Sample records for purification technology applied

  1. Waste water biological purification plants of dairy products industry and energy management

    NASA Astrophysics Data System (ADS)

    Stepanov, Sergey; Solkina, Olga; Stepanov, Alexander; Zhukova, Maria

    2017-10-01

    The paper presents results of engineering and economical comparison of waste water biological purification plants of dairy products industry. Three methods of purification are compared: traditional biological purification with the use of secondary clarifiers and afterpurification through granular-bed filters, biomembrane technology and physical-and-chemical treatment together with biomembrane technology for new construction conditions. The improvement of the biological purification technology using nitro-denitrification and membrane un-mixing of sludge mixture is a promising trend in this area. In these calculations, an energy management which is widely applied abroad was used. The descriptions of the three methods are illustrated with structural schemes. Costs of equipment and production areas are taken from manufacturers’ data. The research is aimed at an engineering and economical comparison of new constructions of waste water purification of dairy products industry. The experiment demonstrates advantages of biomembrane technology in waste water purification. This technology offers prospects of 122 million rubles cost saving during 25 years of operation when compared with of the technology of preparatory reagent flotation and of 13.7 million rubles cost saving compared to the option of traditional biological purification.

  2. SwellGel: an affinity chromatography technology for high-capacity and high-throughput purification of recombinant-tagged proteins.

    PubMed

    Draveling, C; Ren, L; Haney, P; Zeisse, D; Qoronfleh, M W

    2001-07-01

    The revolution in genomics and proteomics is having a profound impact on drug discovery. Today's protein scientist demands a faster, easier, more reliable way to purify proteins. A high capacity, high-throughput new technology has been developed in Perbio Sciences for affinity protein purification. This technology utilizes selected chromatography media that are dehydrated to form uniform aggregates. The SwellGel aggregates will instantly rehydrate upon addition of the protein sample, allowing purification and direct performance of multiple assays in a variety of formats. SwellGel technology has greater stability and is easier to handle than standard wet chromatography resins. The microplate format of this technology provides high-capacity, high-throughput features, recovering milligram quantities of protein suitable for high-throughput screening or biophysical/structural studies. Data will be presented applying SwellGel technology to recombinant 6x His-tagged protein and glutathione-S-transferase (GST) fusion protein purification. Copyright 2001 Academic Press.

  3. Water Purification

    NASA Technical Reports Server (NTRS)

    1992-01-01

    Silver ionization water purification technology was originally developed for Apollo spacecraft. It was later used to cleanse swimming pools and has now been applied to industrial cooling towers and process coolers. Sensible Technologies, Inc. has added two other technologies to the system, which occupies only six square feet. It is manufactured in three capacities, and larger models are custom built on request. The system eliminates scale, corrosion, algae, bacteria and debris, and because of the NASA technology, viruses and waterborne bacteria are also destroyed. Applications include a General Motors cooling tower, amusement parks, ice manufacture and a closed-loop process cooling system.

  4. Supercoiled plasmid DNA purification by integrating membrane technology with a monolithic chromatography.

    PubMed

    Nunes, Catherine; Sousa, Angela; Nunes, José C; Morão, António M; Sousa, Fani; Queiroz, João A

    2014-06-01

    The present study describes the integration of membrane technology with monolithic chromatography to obtain plasmid DNA with high quality. Isolation and clarification of plasmid DNA lysate were first conducted by a microfiltration step, by using a hydrophilic nylon microfiltration membrane, avoiding the need of centrifugation. For the total elimination of the remaining impurities, a suitable purification step is required. Monolithic stationary phases have been successfully applied as an alternative to conventional supports. Thus, the sample recovered from the membrane process was applied into a nongrafted CarbonylDiImidazole disk. Throughout the global procedure, a reduced level of impurities such as proteins and RNA was obtained, and no genomic DNA was detectable in the plasmid DNA sample. The chromatographic process demonstrated an efficient performance on supercoiled plasmid DNA purity and recovery (100 and 84.44%, respectively). Thereby, combining the membrane technology to eliminate some impurities from lysate sample with an efficient chromatographic strategy to purify the supercoiled plasmid DNA arises as a powerful approach for industrial-scale systems aiming at plasmid DNA purification. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Preparative Purification of Recombinant Proteins: Current Status and Future Trends

    PubMed Central

    Saraswat, Mayank; Ravidá, Alessandra; Holthofer, Harry

    2013-01-01

    Advances in fermentation technologies have resulted in the production of increased yields of proteins of economic, biopharmaceutical, and medicinal importance. Consequently, there is an absolute requirement for the development of rapid, cost-effective methodologies which facilitate the purification of such products in the absence of contaminants, such as superfluous proteins and endotoxins. Here, we provide a comprehensive overview of a selection of key purification methodologies currently being applied in both academic and industrial settings and discuss how innovative and effective protocols such as aqueous two-phase partitioning, membrane chromatography, and high-performance tangential flow filtration may be applied independently of or in conjunction with more traditional protocols for downstream processing applications. PMID:24455685

  6. Application of QUAL2K Model to Assess Ecological Purification Technology for a Polluted River

    PubMed Central

    Zhu, Wenting; Niu, Qian; Zhang, Ruibin; Ye, Rui; Qian, Xin; Qian, Yu

    2015-01-01

    Industrialization and urbanization have caused water pollution and ecosystem degradation, especially in urban canals and rivers in China; accordingly, effective water quality improvement programs are needed. In this study, the Tianlai River in Jiangsu, China was taken as a research site, and a combination of ecological purification technologies consisting of biological rope, phytoremediation, and activated carbon were applied in a laboratory-scale study to examine degradation coefficients under dynamic water conditions. Coefficients were then input into the QUAL2K model to simulate various hypothetical scenarios and determine the minimum density of ecological purification combination and hydraulic retention time (HRT) to meet Grade V or IV of the China standard for surface water. The minimum densities for Grade V and IV were 1.6 times and 2 times the experimental density, while the minimum HRTs for Grade V and IV were 2.4 day and 3 day. The results of this study should provide a practical and efficient design method for ecological purification programs. PMID:25689997

  7. Bromelain: an overview of industrial application and purification strategies.

    PubMed

    Arshad, Zatul Iffah Mohd; Amid, Azura; Yusof, Faridah; Jaswir, Irwandi; Ahmad, Kausar; Loke, Show Pau

    2014-09-01

    This review highlights the use of bromelain in various applications with up-to-date literature on the purification of bromelain from pineapple fruit and waste such as peel, core, crown, and leaves. Bromelain, a cysteine protease, has been exploited commercially in many applications in the food, beverage, tenderization, cosmetic, pharmaceutical, and textile industries. Researchers worldwide have been directing their interest to purification strategies by applying conventional and modern approaches, such as manipulating the pH, affinity, hydrophobicity, and temperature conditions in accord with the unique properties of bromelain. The amount of downstream processing will depend on its intended application in industries. The breakthrough of recombinant DNA technology has facilitated the large-scale production and purification of recombinant bromelain for novel applications in the future.

  8. Use of naturally growing aquatic plants for wastewater purification.

    PubMed

    Zimmels, Y; Kirzhner, F; Roitman, S

    2004-01-01

    This paper examines potential uses of naturally growing aquatic plants for wastewater purification. These plants enhance the removal of pollutants by consuming part of them in the form of plant nutrients. This applies to urban and agricultural wastewater, in particular, where treatment units of different sizes can be applied at the pollution source. The effectiveness of wastewater purification by different plants was tested on laboratory and pilot scales. The growth rate of the plants was related to the wastewater content in the water. Batch and semicontinuous experiments verified that the plants are capable of decreasing all tested indicators for water quality to levels that permit the use of the purified water for irrigation. This applies to biochemical oxygen demand (BOD), chemical oxygen demand, total suspended solids. pH, and turbidity. In specific cases, the turbidity reached the level of drinking water. Comparison of BOD concentrations with typical levels in water treatment facilities across the country indicates the effectiveness of water purification with plants. A major effect of treatment with plants was elimination of the disturbing smell from the wastewater. It is shown that mixtures of wastewater and polluted water from the Kishon River are amenable in varying degrees to treatment by the plants. The higher the wastewater content in the mixture, the more effective the treatment by the plants. In this context, a scheme for rehabilitation and restoration of the Kishon River is presented and technical and economical aspects of the purification technology are considered.

  9. Clarification of vaccines: An overview of filter based technology trends and best practices.

    PubMed

    Besnard, Lise; Fabre, Virginie; Fettig, Michael; Gousseinov, Elina; Kawakami, Yasuhiro; Laroudie, Nicolas; Scanlan, Claire; Pattnaik, Priyabrata

    2016-01-01

    Vaccines are derived from a variety of sources including tissue extracts, bacterial cells, virus particles, recombinant mammalian, yeast and insect cell produced proteins and nucleic acids. The most common method of vaccine production is based on an initial fermentation process followed by purification. Production of vaccines is a complex process involving many different steps and processes. Selection of the appropriate purification method is critical to achieving desired purity of the final product. Clarification of vaccines is a critical step that strongly impacts product recovery and subsequent downstream purification. There are several technologies that can be applied for vaccine clarification. Selection of a harvesting method and equipment depends on the type of cells, product being harvested, and properties of the process fluids. These techniques include membrane filtration (microfiltration, tangential-flow filtration), centrifugation, and depth filtration (normal flow filtration). Historically vaccine harvest clarification was usually achieved by centrifugation followed by depth filtration. Recently membrane based technologies have gained prominence in vaccine clarification. The increasing use of single-use technologies in upstream processes necessitated a shift in harvest strategies. This review offers a comprehensive view on different membrane based technologies and their application in vaccine clarification, outlines the challenges involved and presents the current state of best practices in the clarification of vaccines. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. Application of visual basic in high-throughput mass spectrometry-directed purification of combinatorial libraries.

    PubMed

    Li, B; Chan, E C Y

    2003-01-01

    We present an approach to customize the sample submission process for high-throughput purification (HTP) of combinatorial parallel libraries using preparative liquid chromatography electrospray ionization mass spectrometry. In this study, Visual Basic and Visual Basic for Applications programs were developed using Microsoft Visual Basic 6 and Microsoft Excel 2000, respectively. These programs are subsequently applied for the seamless electronic submission and handling of data for HTP. Functions were incorporated into these programs where medicinal chemists can perform on-line verification of the purification status and on-line retrieval of postpurification data. The application of these user friendly and cost effective programs in our HTP technology has greatly increased our work efficiency by reducing paper work and manual manipulation of data.

  11. Current Progress of Capacitive Deionization for Removal of Pollutant Ions

    NASA Astrophysics Data System (ADS)

    Gaikwad, Mahendra S.; Balomajumder, Chandrajit

    2016-08-01

    A mini review of a recently developing water purification technology capacitive deionization (CDI) applied for removal of pollutant ions is provided. The current progress of CDI for removal of different pollutant ions such as arsenic, fluoride, boron, phosphate, lithium, copper, cadmium, ferric, and nitrate ions is presented. This paper aims at motivating new research opportunities in capacitive deionization technology for removal of pollutant ions from polluted water.

  12. Removal of dioxins and furans from flue gases by non-flammable adsorbents in a fixed bed.

    PubMed

    Fell, H J; Tuczek, M

    1998-01-01

    The presented adsorption--process KOMBISORBON is applied for high efficient off-gas purification, preferably of polychlorinated dioxins and furans from off-gas of incineration plants, which are generated, when these are operated under unfavourable conditions [2]. This off-gas purification process complies with german laws, which limit the concentration of these substances to less than 0.1 ng toxicity equivalents (TE) per cubic metre of gas [1]. The adsorbent, the adsorption process and its plant concept (fixed bed) is described in detail including economics and obtained operation results. Alternative removal technologies are briefly outlined.

  13. Cell purification: a new challenge for biobanks.

    PubMed

    Almeida, Maria; García-Montero, Andres C; Orfao, Alberto

    2014-01-01

    Performing '-omics' analyses on heterogeneous biological tissue samples, such as blood or bone marrow, can lead to biased or even erroneous results, particularly when the targeted cells and/or molecules are present at relatively low percentages/amounts. In such cases, whole sample analysis will most probably dilute and mask the features of the cell and/or molecules of interest, and this will negatively impact the results and their interpretation. Therefore, frequently it is critically important to have well-characterized and high-quality purified cell populations for the reliable detection of subtle variations in their specific features, such as gene expression profile, protein expression pattern and metabolic status. Biobanks are technological platforms which aim to provide researchers access to a large number of high-quality biological samples and their associated data, particularly to support high-quality scientific and clinical research projects, and such projects will benefit enormously by having access to high-quality purified cell populations or their biological components (e.g. DNA, RNA, proteins). Therefore, a clear opportunity exists for preparative cell sorting techniques in biobanks. Although multiple different cell purification approaches exist or are under development (e.g. cell purification techniques based on cell adherence, density and/or cell size properties, methods based on antibody binding as well as new lab-on-a-chip purification techniques), the choice for a specific technology depends on multiple variables, including cell recovery, purity and yield, among others. In addition, most cell purification approaches are not well suited for high-throughput (HT) purification of multiple cell populations coexisting in a sample. Here we review the most (currently) used cell sorting methods that may be applied for sample preparation in biobanks. For the different approaches, technical considerations about their advantages and limitations are highlighted, and the requirements to be met by a HT cell sorting technology to be used in biobanks are also discussed.

  14. Arsanilic acid modified superparamagnetic iron oxide nanoparticles for Purification of alkaline phosphatase from hen's egg yolk.

    PubMed

    Farzi-Khajeh, Hamed; Safa, Kazem D; Dastmalchi, Siavoush

    2017-09-01

    Recent studies of magnetic carrier technology have focused on its applications in separation and purification technologies, due to easy separation of the target from the reaction medium by applying an external magnetic field. In the present study, Fe 3 O 4 superparamagnetic nanoparticles were prepared to utilize a chemical co-precipitation method, then the surfaces of the nanoparticles were modified with arsanilic acid derivatives which were used as the specific nanocarriers for the affinity purification of alkaline phosphatase from the hen's egg yolk. The six different types of magnetic nanocarriers with varied lengths of the linkers were obtained. All samples were characterized step by step and validated using FTIR, SEM, EDX, VSM and XRD analysis methods As the results were shown, the use of inflexible tags with long linkers on the surface of the nanocarrier could lead to better results for separation of alkaline phosphatase from the hen's egg yolk with 76.2% recovery and 1361.7-fold purification. The molecular weight of the purified alkaline phosphatase was estimated to be 68kDa by SDS-PAGE. The results of this study showed that the novel magnetic nanocarriers were capable of purifying alkaline phosphatase in a practically time and cost effective way. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. On-chip purification and detection of hepatitis C virus RNA from human plasma.

    PubMed

    Vaghi, V; Potrich, C; Pasquardini, L; Lunelli, L; Vanzetti, L; Ebranati, E; Lai, A; Zehender, G; Mombello, D; Cocuzza, M; Pirri, C F; Pederzolli, C

    2016-01-01

    Hepatitis C virus (HCV) is one of the main causes of chronic liver disease worldwide. The diagnosis and monitoring of HCV infection is a crucial need in the clinical management. The conventional diagnostic technologies are challenged when trying to address molecular diagnostics, especially because they require a complex and time-consuming sample preparation phase. Here, a new concept based on surface functionalization was applied to viral RNA purification: first of all polydimethylsiloxane (PDMS) flat surfaces were modified to hold RNA adsorption. After a careful chemical and morphological analysis of the modified surfaces, the functionalization protocols giving the best RNA adsorbing surfaces were applied to PDMS microdevices. The functionalized microdevices were then used for RNA purification from HCV infected human plasma samples. RNA purification and RT were successfully performed in the same microdevice chamber, saving time of analysis, reagents, and labor. The PCR protocol for HCV cDNA amplification was also implemented in the microdevice, demonstrating that the entire process of HCV analysis, from plasma to molecular readout, could be performed on-chip. Not only HCV but also other microdevice-based viral RNA detection could therefore result in a successful Point-of-Care (POC) diagnostics for resource-limited settings. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. [Pilot-scale purification of lipopeptide from marine-derived Bacillus marinus].

    PubMed

    Gu, Kangbo; Guan, Cheng; Xu, Jiahui; Li, Shulan; Luo, Yuanchan; Shen, Guomin; Zhang, Daojing; Li, Yuanguang

    2016-11-25

    This research was aimed at establishing the pilot-scale purification technology of lipopeptide from marine-derived Bacillus marinus. We studied lipopeptide surfactivity interferences on scale-up unit technologies including acid precipitation, methanol extraction, solvent precipitation, salting out, extraction, silica gel column chromatography and HZ806 macroporous absorption resin column chromatography. Then, the unit technologies were combined in a certain order, to remove the impurities gradually, and to gain purified lipopeptide finally, with high recovery rate throughout the whole process. The novel pilot-scale purification technology could effectively isolate and purify lipopeptide with 87.51% to 100% purity in hectograms from 1 ton of Bacillus marinus B-9987 fermentation broth with more than 81.73% recovery rate. The first practical hectogram production of highly purified lipopeptide derived from Bacillus marinus was achieved. With this new purification method, using complex media became possible in fermentation process to reduce the fermentation cost and scale-up the purification for lipopeptide production. For practicability and economy, foaming problem resulting from massive water evaporation was avoided in this technology.

  17. [Purification Technology Optimization for Saponins from Ziziphi Spinosae Semen with Macroporous Adsorption Resin by Box-Behnken Design-Response Surface Methodology].

    PubMed

    Zhao, Hui-ru; Ren, Zao; Liu, Chun-ye

    2015-04-01

    To compare the purification effect of saponins from Ziziphi Spinosae Semen with different types of macroporous adsorption resin, and to optimize its purification technology. The type of macroporous resins was optimized by static adsorption method. The optimum technological conditions of saponins from Ziziphi Spinosae Semen was screened by single factor test and Box-Behnken Design-Response Surface Methodology. AB-8 macroporous resin had better purification effect of total saponins than other resins, optimum technological parameters were as follows: column height-diameter ratio was 5: 1, the concentration of sample solution was 2. 52 mg/mL, resin adsorption quantity was 8. 915 mg/g, eluted by 3 BV water, flow rate of adsorption and elution was 2 BV/h, elution solvent was 75% ethanol, elution solvent volume was 5 BV. AB-8 macroporous resin has a good purification effect on jujuboside A. The optimized technology is stable and feasible.

  18. Improved technique that allows the performance of large-scale SNP genotyping on DNA immobilized by FTA technology.

    PubMed

    He, Hongbin; Argiro, Laurent; Dessein, Helia; Chevillard, Christophe

    2007-01-01

    FTA technology is a novel method designed to simplify the collection, shipment, archiving and purification of nucleic acids from a wide variety of biological sources. The number of punches that can normally be obtained from a single specimen card are often however, insufficient for the testing of the large numbers of loci required to identify genetic factors that control human susceptibility or resistance to multifactorial diseases. In this study, we propose an improved technique to perform large-scale SNP genotyping. We applied a whole genome amplification method to amplify DNA from buccal cell samples stabilized using FTA technology. The results show that using the improved technique it is possible to perform up to 15,000 genotypes from one buccal cell sample. Furthermore, the procedure is simple. We consider this improved technique to be a promising methods for performing large-scale SNP genotyping because the FTA technology simplifies the collection, shipment, archiving and purification of DNA, while whole genome amplification of FTA card bound DNA produces sufficient material for the determination of thousands of SNP genotypes.

  19. Scale-up of hydrophobin-assisted recombinant protein production in tobacco BY-2 suspension cells.

    PubMed

    Reuter, Lauri J; Bailey, Michael J; Joensuu, Jussi J; Ritala, Anneli

    2014-05-01

    Plant suspension cell cultures are emerging as an alternative to mammalian cells for production of complex recombinant proteins. Plant cell cultures provide low production cost, intrinsic safety and adherence to current regulations, but low yields and costly purification technology hinder their commercialization. Fungal hydrophobins have been utilized as fusion tags to improve yields and facilitate efficient low-cost purification by surfactant-based aqueous two-phase separation (ATPS) in plant, fungal and insect cells. In this work, we report the utilization of hydrophobin fusion technology in tobacco bright yellow 2 (BY-2) suspension cell platform and the establishment of pilot-scale propagation and downstream processing including first-step purification by ATPS. Green fluorescent protein-hydrophobin fusion (GFP-HFBI) induced the formation of protein bodies in tobacco suspension cells, thus encapsulating the fusion protein into discrete compartments. Cultivation of the BY-2 suspension cells was scaled up in standard stirred tank bioreactors up to 600 L production volume, with no apparent change in growth kinetics. Subsequently, ATPS was applied to selectively capture the GFP-HFBI product from crude cell lysate, resulting in threefold concentration, good purity and up to 60% recovery. The ATPS was scaled up to 20 L volume, without loss off efficiency. This study provides the first proof of concept for large-scale hydrophobin-assisted production of recombinant proteins in tobacco BY-2 cell suspensions. © 2013 Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.

  20. Membrane Technologies in Wine Industry: An Overview.

    PubMed

    El Rayess, Youssef; Mietton-Peuchot, Martine

    2016-09-09

    Membrane processes are increasingly reported for various applications in wine industry such as microfiltration, electrodialysis, and reverse osmosis, but also emerging processes as bipolar electrodialysis and membrane contactor. Membrane-based processes are playing a critical role in the field of separation/purification, clarification, stabilization, concentration, and de-alcoholization of wine products. They begin to be an integral part of the winemaking process. This review will provide an overview of recent developments, applications, and published literature in membrane technologies applied in wine industry.

  1. Discussion on runoff purification technology of highway bridge deck based on water quality safety

    NASA Astrophysics Data System (ADS)

    Tan, Sheng-guang; Liu, Xue-xin; Zou, Guo-ping; Xiong, Xin-zhu; Tao, Shuang-cheng

    2018-06-01

    Aiming at the actual problems existing, including a poor purification effect of highway bridge runoff collection and treatment system across sensitive water and necessary manual emergency operation, three kinds of technology, three pools system of bridge runoff purification, the integral pool of bridge runoff purification and ecological planting tank, are put forward by optimizing the structure of purification unit and system setting. At the same time, we come up with an emergency strategy for hazardous material leakage basing on automatic identification and remote control of traffic accidents. On the basis of combining these with the optimized pool structure, sensitive water safety can be guaranteed and water pollution, from directly discharging of bridge runoff, can be decreased. For making up for the shortages of green highway construction technology, the technique has important reference value.

  2. Effect of HEH[EHP] impurities on the ALSEP solvent extraction process

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Holfeltz, Vanessa E.; Campbell, Emily L.; Peterman, Dean R.

    In solvent extraction processes, organic phase impurities can negatively impact separation factors, hydrolytic performance, and overall system robustness. This affects the process-level viability of a separation concept and necessitates knowledge of the behavior and mechanisms to control impurities in the solvent. The most widespread way through which impurities are introduced into a system is through impure extractants and/or diluents used to prepare the solvent, and often development of new purification schemes to achieve the desired level of purity is needed. In this work, the acidic extractant, 2-ethylhexylphosphonic acid mono-2-ethylhexyl ester (HEH[EHP])—proposed for application in extractive processes aimed at separating trivalentmore » minor actinides from lanthanides and other fission products—is characterized with respect to its common impurities and their impact on Am(III) stripping in the Actinide Lanthanide SEParation (ALSEP) system. To control impurities in HEH[EHP], existing purification technologies commonly applied for the acidic organophosphorus reagents are reviewed, and a new method specific to HEH[EHP] purification is presented.« less

  3. Impulse powerful UV-radiation source pumped by the sublight ionization waves for the bacteriological disinfection of water

    NASA Astrophysics Data System (ADS)

    Filiouguine, Igor V.; Kostiouchenko, S. V.; Koudryavtsev, N. N.; Vasilyak, Leonid M.; Yakimenko, A. V.

    1993-11-01

    The bacteriological disinfective action of UV-radiation is well known. The pioneer work on UV-radiation used for bacteriological disinfection of waste water was made in 1910. Because of the high cost and low living time of the UV-radiation sources, the alternative technique for waste water purification by chlorine introducing was spread out. During the second stage of the UV purification development, beginning in approximately 1970, the interest for bacteriological cleaning of water, increased again. Two reasons were responsible for this event: first, the significant improvement of technology and design of UV-bacteriological purificators, and second, recognition of the serious danger of chlorine compounds introduced into water under purification because of the toxicity of these compounds. Further investigations gave excellent results in the creation and industrial applications of UV- bacteriological purificators. Now we can see a rapid development of industrial technology in UV-purification of drinking and waste waters.

  4. Characterization of the diatomite binding domain in the ribosomal protein L2 from E. coli and functions as an affinity tag.

    PubMed

    Li, Junhua; Zhang, Yang; Yang, Yanjun

    2013-03-01

    The ribosomal protein L2, a constituent protein of the 50S large ribosomal subunit, can be used as Si-tag using silica particles for the immobilization and purification of recombinant proteins (Ikeda et al. (Protein Expr Purif 71:91-95, 2010); Taniguchi et al. (Biotechnol Bioeng 96:1023-1029, 2007)). We applied a diatomite powder, a sedimentary rock mainly composed with diatoms silica, as an affinity solid phase and small ubiquitin-like modifier (SUMO) technology to release a target protein from the solid phase. The L2 (203-273) was the sufficient region for the adsorption of ribosomal protein L2 on diatomite. We comparatively analyzed the different adsorption properties of the two deleted proteins of L2 (L2 (1-60, 203-273) and L2 (203-273)) on diatomite. The time required to reach adsorption equilibrium of L2 (203-273) fusion protein on diatomite was shorter than that of L2 (1-60, 203-273) fusion protein. The maximum adsorption capacity of L2 (203-273) fusion protein was larger than that of L2 (1-60, 203-273) fusion protein. In order to study whether the L2 (203-273) can function as an affinity purification tag, SUMO was introduced as one specific protease cleavage site between the target protein and the purification tags. The L2 (203-273) and SUMO fusion protein purification method was tested using enhanced green fluorescent protein as a model protein; the result shows that the purification performance of this affinity purification method was good. The strong adsorption characteristic of L2 (203-273) on diatomite also provides a potential protein fusion tag for the immobilization of enzyme.

  5. Development of Ultrafiltration Membrane-Separation Technology for Energy-Efficient Water Treatment and Desalination Process

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Yim, Woosoon; Bae, Chulsung

    The growing scarcity of fresh water is a major political and economic challenge in the 21st century. Compared to thermal-based distillation technique of water production, pressure driven membrane-based water purification process, such as ultrafiltration (UF), nanofiltration (NF) and reverse osmosis (RO), can offer more energy-efficient and environmentally friendly solution to clean water production. Potential applications also include removal of hazardous chemicals (i.e., arsenic, pesticides, organics) from water. Although those membrane-separation technologies have been used to produce drinking water from seawater (desalination) and non-traditional water (i.e., municipal wastewater and brackish groundwater) over the last decades, they still have problems in ordermore » to be applied in large-scale operations. Currently, a major huddle of membrane-based water purification technology for large-scale commercialization is membrane fouling and its resulting increases in pressure and energy cost of filtration process. Membrane cleaning methods, which can restore the membrane properties to some degree, usually cause irreversible damage to the membranes. Considering that electricity for creating of pressure constitutes a majority of cost (~50%) in membrane-based water purification process, the development of new nano-porous membranes that are more resistant to degradation and less subject to fouling is highly desired. Styrene-ethylene/butylene-styrene (SEBS) block copolymer is one of the best known block copolymers that induces well defined morphologies. Due to the polarity difference of aromatic styrene unit and saturated ethylene/butylene unit, these two polymer chains self-assemble each other and form different phase-separated morphologies depending on the ratios of two polymer chain lengths. Because the surface of SEBS is hydrophobic which easily causes fouling of membrane, incorporation of ionic group (e,g, sulfonate) to the polymer is necessary to reduces fouling. Recently, sulfonated SEBS became commercially available and has been extensively explored for membrane-mediated water purification technology. The sulfonated block copolymer creates a well developed nano-sale phase-separated morphologies composed of hydrophilic domains (sulfonated polystyrene) and hydrophobic domains (polyethylene/polybutylene). The hydrophilic domains determines transport properties (water transport, salt and/or ion rejection, etc) and the hydrophobic domains provides mechanical stability of the membrane. Unfortunately, a high degree of sulfonation of SEBS induces excessive swelling and deterioration of mechanical stability of the membrane. In an effort to develop robust polymeric membrane materials for water purification technology, phosphonic acid-functionalized SEBS membranes are investigated during this report period. In compare to sulfonated polymers, the corresponding phosphonated polymers are known to swell less because of the formation of extensive hydrogen bonding networks between phosphonates. In addition to the expected better mechanical stability, phosphonated polymers has another advantage over sulfonated polymers for the use water purification membrane; each phosphonate can accommodate two ions while each sulfonate accommodates only one ion. Membrane properties (ion type, ionic density, etc) of new membranes will be studied and their separation performance will be evaluated in water purification and desalination process. Through systematic study of the relationship of chemical structure–surface property–membrane performance, we aim to better understand the nature of membrane fouling and develop more fouling-resistant water purification membranes. The basic understanding of this relationship will lead to the development of advanced membrane materials which can offer a solution to environmentally sustainable production of fresh water.« less

  6. Evaluation of immobilized metal membrane affinity chromatography for purification of an immunoglobulin G1 monoclonal antibody.

    PubMed

    Serpa, Gisele; Augusto, Elisabeth Fátima Pires; Tamashiro, Wirla Maria Silva Cunha; Ribeiro, Mariana Borçoe; Miranda, Everson Alves; Bueno, Sônia Maria Alves

    2005-02-25

    The large scale production of monoclonal antibodies (McAbs) has gaining increased relevance with the development of the hybridoma cell culture in bioreactors creating a need for specific efficient bioseparation techniques. Conventional fixed bead affinity adsorption commonly applied for McAbs purification has the drawback of low flow rates and colmatage. We developed and evaluated a immobilized metal affinity chromatographies (IMAC) affinity membrane for the purification of anti-TNP IgG(1) mouse McAbs. We immobilized metal ions on a poly(ethylene vinyl alcohol) hollow fiber membrane (Me(2+)-IDA-PEVA) and applied it for the purification of this McAbs from cell culture supernatant after precipitation with 50% saturation of ammonium sulphate. The purity of IgG(1) in the eluate fractions was high when eluted from Zn(2+) complex. The anti-TNP antibody could be eluted under conditions causing no loss of antigen binding capacity. The purification procedure can be considered as an alternative to the biospecific adsorbent commonly applied for mouse IgG(1) purification, the protein G-Sepharose.

  7. Experimental Study on the Plasma Purification for Diesel Engine Exhaust Gas

    NASA Astrophysics Data System (ADS)

    Chen, Jing; Zu, Kan; Wang, Mei

    2018-02-01

    It is known that the use of ternary catalysis is capable of significantly reducing the emission of pollutants from petrol vehicles. However, the disadvantages such as the temperature and other limitations make it unsuitable for diesel engines. The plasma-assisted catalyst technology has been applied in dealing with the diesel exhaust in the experiment in order to do further research on the effects of plasma in exhaust processing. The paper not only includes the experimental observation on the change of particle concentration after the operation of purification device, but also builds the kinetic model of chemical reactions to simulate the reactions of nitrogen oxides in plasma through using the software of Matlab, then compares the calculation results with experimental samples and finally gets some useful conclusions in practice.

  8. Photocatalytic materials and technologies for air purification.

    PubMed

    Ren, Hangjuan; Koshy, Pramod; Chen, Wen-Fan; Qi, Shaohua; Sorrell, Charles Christopher

    2017-03-05

    Since there is increasing concern for the impact of air quality on human health, the present work surveys the materials and technologies for air purification using photocatalytic materials. The coverage includes (1) current photocatalytic materials for the decomposition of chemical contaminants and disinfection of pathogens present in air and (2) photocatalytic air purification systems that are used currently and under development. The present work focuses on five main themes. First, the mechanisms of photodegradation and photodisinfection are explained. Second, system designs for photocatalytic air purification are surveyed. Third, the photocatalytic materials used for air purification and their characteristics are considered, including both conventional and more recently developed photocatalysts. Fourth, the methods used to fabricate these materials are discussed. Fifth, the most significant coverage is devoted to materials design strategies aimed at improving the performance of photocatalysts for air purification. The review concludes with a brief consideration of promising future directions for materials research in photocatalysis. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Affinity approaches in RNAi-based therapeutics purification.

    PubMed

    Pereira, Patrícia; Queiroz, João A; Figueiras, Ana; Sousa, Fani

    2016-05-15

    The recent investigation on RNA interference (RNAi) related mechanisms and applications led to an increased awareness of the importance of RNA in biology. Nowadays, RNAi-based technology has emerged as a potentially powerful tool for silencing gene expression, being exploited to develop new therapeutics for treating a vast number of human disease conditions, as it is expected that this technology can be translated onto clinical applications in a near future. This approach makes use of a large number of small (namely short interfering RNAs, microRNAs and PIWI-interacting RNAs) and long non-coding RNAs (ncRNAs), which are likely to have a crucial role as the next generation therapeutics. The commercial and biomedical interest in these RNAi-based therapy applications have fostered the need to develop innovative procedures to easily and efficiently purify RNA, aiming to obtain the final product with high purity degree, good quality and biological activity. Recently, affinity chromatography has been applied to ncRNAs purification, in view of the high specificity. Therefore, this article intends to review the biogenesis pathways of regulatory ncRNAs and also to discuss the most significant and recent developments as well as applications of affinity chromatography in the challenging task of purifying ncRNAs. In addition, the importance of affinity chromatography in ncRNAs purification is addressed and prospects for what is forthcoming are presented. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Water Purification Product

    NASA Technical Reports Server (NTRS)

    2004-01-01

    Ecomaster, an affiliate of BioServe Space Technologies, this PentaPure technology has been used to purify water for our nation's Space Shuttle missions since 1981. WTC-Ecomaster of Mirneapolis, Minnesota manufactures water purification systems under the brand name PentaPure (TM). BioServe researcher Dr. George Marchin, of Kansas State University, first demonstrated the superiority of this technology and licensed it to WTC. Marchin continues to perform microgravity research in the development of new technologies for the benefit of life on Earth.

  11. Biochar-based functional materials in the purification of agricultural wastewater: Fabrication, application and future research needs.

    PubMed

    Wei, Dongning; Li, Bingyu; Huang, Hongli; Luo, Lin; Zhang, Jiachao; Yang, Yuan; Guo, Jiajun; Tang, Lin; Zeng, Guangming; Zhou, Yaoyu

    2018-04-01

    Nowadays, agricultural contamination is becoming more and more serious due to the rapid growth of agricultural industry, which discharged antibiotics, pesticides or toxic metals into farmlands. A large number of researchers have applied biochar-based functional materials to the treatment of agricultural wastewater contamination. Meanwhile, biochar has also proved to be a very promising and effective technology in water purification field due to its various beneficial properties (e.g., cost effective, high specific surface area, and surface reactive groups). The focus of this review is to highlight the fabrication methods and application of biochar-based functional materials with the removal of different agricultural contaminants, and discuss the underlying mechanisms. However, the application of biochar-based functional materials is currently under its infancy, with the main hindrance is identified as the gap between laboratory scale and field application, immaturity of engineered biochar production technologies, and lack of quality standards. In order to fill these knowledge gaps, more efforts should be made to pay for the relevant research in future studies. Copyright © 2018 Elsevier Ltd. All rights reserved.

  12. Air and Water System (AWS) Design and Technology Selection for the Vision for Space Exploration

    NASA Technical Reports Server (NTRS)

    Jones, Harry; Kliss, Mark

    2005-01-01

    This paper considers technology selection for the crew air and water recycling systems to be used in long duration human space exploration. The specific objectives are to identify the most probable air and water technologies for the vision for space exploration and to identify the alternate technologies that might be developed. The approach is to conduct a preliminary first cut systems engineering analysis, beginning with the Air and Water System (AWS) requirements and the system mass balance, and then define the functional architecture, review the International Space Station (ISS) technologies, and discuss alternate technologies. The life support requirements for air and water are well known. The results of the mass flow and mass balance analysis help define the system architectural concept. The AWS includes five subsystems: Oxygen Supply, Condensate Purification, Urine Purification, Hygiene Water Purification, and Clothes Wash Purification. AWS technologies have been evaluated in the life support design for ISS node 3, and in earlier space station design studies, in proposals for the upgrade or evolution of the space station, and in studies of potential lunar or Mars missions. The leading candidate technologies for the vision for space exploration are those planned for Node 3 of the ISS. The ISS life support was designed to utilize Space Station Freedom (SSF) hardware to the maximum extent possible. The SSF final technology selection process, criteria, and results are discussed. Would it be cost-effective for the vision for space exploration to develop alternate technology? This paper will examine this and other questions associated with AWS design and technology selection.

  13. Nanodiamonds of Laser Synthesis for Biomedical Applications.

    PubMed

    Perevedentseva, E; Peer, D; Uvarov, V; Zousman, B; Levinson, O

    2015-02-01

    In recent decade detonation nanodiamonds (DND), discovered 50 years ago and used in diverse technological processes, have been actively applied in biomedical research as a drug and gene delivery carrier, a contrast agent for bio-imaging and diagnostics and an adsorbent for protein separation and purification. In this work we report about nanodiamonds of high purity produced by laser assisted technique, compare them with DND and consider the prospect and advantages of their use in the said applications.

  14. Single-step affinity purification of enzyme biotherapeutics: a platform methodology for accelerated process development.

    PubMed

    Brower, Kevin P; Ryakala, Venkat K; Bird, Ryan; Godawat, Rahul; Riske, Frank J; Konstantinov, Konstantin; Warikoo, Veena; Gamble, Jean

    2014-01-01

    Downstream sample purification for quality attribute analysis is a significant bottleneck in process development for non-antibody biologics. Multi-step chromatography process train purifications are typically required prior to many critical analytical tests. This prerequisite leads to limited throughput, long lead times to obtain purified product, and significant resource requirements. In this work, immunoaffinity purification technology has been leveraged to achieve single-step affinity purification of two different enzyme biotherapeutics (Fabrazyme® [agalsidase beta] and Enzyme 2) with polyclonal and monoclonal antibodies, respectively, as ligands. Target molecules were rapidly isolated from cell culture harvest in sufficient purity to enable analysis of critical quality attributes (CQAs). Most importantly, this is the first study that demonstrates the application of predictive analytics techniques to predict critical quality attributes of a commercial biologic. The data obtained using the affinity columns were used to generate appropriate models to predict quality attributes that would be obtained after traditional multi-step purification trains. These models empower process development decision-making with drug substance-equivalent product quality information without generation of actual drug substance. Optimization was performed to ensure maximum target recovery and minimal target protein degradation. The methodologies developed for Fabrazyme were successfully reapplied for Enzyme 2, indicating platform opportunities. The impact of the technology is significant, including reductions in time and personnel requirements, rapid product purification, and substantially increased throughput. Applications are discussed, including upstream and downstream process development support to achieve the principles of Quality by Design (QbD) as well as integration with bioprocesses as a process analytical technology (PAT). © 2014 American Institute of Chemical Engineers.

  15. [Studies on preparative technology and quantitative determination for extracts of total saponin in roof of Panax japonicus].

    PubMed

    He, Yu-min; Lu, Ke-ming; Yuan, Ding; Zhang, Chang-cheng

    2008-11-01

    To explore the optimum extraction and purification condition of the total saponins in the root of Panax japonicus (RPJ), and establish its quality control methods. Designed L16 (4(5)) orthogonal test with the extraction rate of total saponins as index, to determine the rational extraction process, and the techniques of water-saturated n-butanol extraction and acetone precipitation were applied to purify the alcohol extract of RPJ. Total saponins were detected by spectrophotometry and its triterpenoidal sapogenin oleanolic acid detected by HPLC. The optimum conditions of total saponins from RPJ was as follows: the material was pulverized, dipped in 60% ethanol aqueous solution as extract solvent at 10 times of volume, and refluxed 3 times for 3 h each time. Extractant of water-saturated n-butanol with extraction times of 3 and precipitant of acetone with precipitation amount of 4-5 times were included in the purification process, which would obtain the quality products. The content of total saponins could reach to 83.48%, and oleanolic acid to 38.30%. The optimized preparative technology is stable, convenient and practical. The extract rate of RPJ was high and steady with this technology, which provided new evidence for industrializing production of the plant and developing new drug.

  16. Phytofilter - environmental friendly solution for purification of surface plate from urbanized territories

    NASA Astrophysics Data System (ADS)

    Ruchkinova, O.; Shchuckin, I.

    2017-06-01

    Its proved, that phytofilters are environmental friendly solution of problem of purification of surface plate from urbanized territories. Phytofilters answer the nowadays purposes to systems of purification of land drainage. The main problem of it is restrictions, connecter with its use in the conditions of cold temperature. Manufactured a technology and mechanism, which provide a whole-year purification of surface plate and its storage. Experimentally stated optimal makeup of filtering load: peat, zeolite and sand in per cent of volume, which provides defined hydraulic characteristics. Stated sorbate and ion-selective volume of complex filtering load of ordered composition in dynamic conditions. Estimated dependences of exit concentrations of oil products and heavy metals on temperature by filtering through complex filtering load of ordered composition. Defined effectiveness of purification at phytofiltering installation. Fixed an influence of embryophytes on process of phytogeneration and capacity of filtering load. Recommended swamp iris, mace reed and reed grass. Manufactured phytofilter calculation methodology. Calculated economic effect from use of phytofiltration technology in comparison with traditional block-modular installations.

  17. New research on bioregenerative air/water purification systems

    NASA Technical Reports Server (NTRS)

    Johnson, Anne H.; Ellender, R. D.; Watkins, Paul J.

    1991-01-01

    For the past several years, air and water purification systems have been developed and used. This technology is based on the combined activities of plants and microorganisms as they function in a natural environment. More recently, researchers have begun to address the problems associated with indoor air pollution. Various common houseplants are currently being evaluated for their abilities to reduce concentrations of volatile organic compounds (VOCS) such as formaldehyde and benzene. With development of the Space Exploration Initiative, missions will increase in duration, and problems with resupply necessitates implementation of regenerative technology. Aspects of bioregenerative technology have been included in a habitat known as the BioHome. The ultimate goal is to use this technology in conjunction with physicochemical systems for air and water purification within closed systems. This study continued the risk assessment of bioregenerative technology with emphasis on biological hazards. In an effort to evaluate the risk for human infection, analyses were directed at enumeration of fecal streptococci and enteric viruses with the BioHome waste water treatment system.

  18. Analysis and evaluation in the production process and equipment area of the low-cost solar array project

    NASA Technical Reports Server (NTRS)

    Goldman, H.; Wolf, M.

    1979-01-01

    Analyses of slicing processes and junction formation processes are presented. A simple method for evaluation of the relative economic merits of competing process options with respect to the cost of energy produced by the system is described. An energy consumption analysis was developed and applied to determine the energy consumption in the solar module fabrication process sequence, from the mining of the SiO2 to shipping. The analysis shows that, in current technology practice, inordinate energy use in the purification step, and large wastage of the invested energy through losses, particularly poor conversion in slicing, as well as inadequate yields throughout. The cell process energy expenditures already show a downward trend based on increased throughput rates. The large improvement, however, depends on the introduction of a more efficient purification process and of acceptable ribbon growing techniques.

  19. Using Haloarcula marismortui Bacteriorhodopsin as a Fusion Tag for Enhancing and Visible Expression of Integral Membrane Proteins in Escherichia coli

    PubMed Central

    Hsu, Min-Feng; Yu, Tsung-Fu; Chou, Chia-Cheng; Fu, Hsu-Yuan; Yang, Chii-Shen; Wang, Andrew H. J.

    2013-01-01

    Membrane proteins are key targets for pharmacological intervention because of their vital functions. Structural and functional studies of membrane proteins have been severely hampered because of the difficulties in producing sufficient quantities of properly folded and biologically active proteins. Here we generate a high-level expression system of integral membrane proteins in Escherichia coli by using a mutated bacteriorhodopsin (BR) from Haloarcula marismortui (HmBRI/D94N) as a fusion partner. A purification strategy was designed by incorporating a His-tag on the target membrane protein for affinity purification and an appropriate protease cleavage site to generate the final products. The fusion system can be used to detect the intended target membrane proteins during overexpression and purification either with the naked eye or by directly monitoring their characteristic optical absorption. In this study, we applied this approach to produce two functional integral membrane proteins, undecaprenyl pyrophosphate phosphatase and carnitine/butyrobetaine antiporter with significant yield enhancement. This technology could facilitate the development of a high-throughput strategy to screen for conditions that improve the yield of correctly folded target membrane proteins. Other robust BRs can also be incorporated in this system. PMID:23457558

  20. Choices of capture chromatography technology in antibody manufacturing processes.

    PubMed

    DiLeo, Michael; Ley, Arthur; Nixon, Andrew E; Chen, Jie

    2017-11-15

    The capture process employed in monoclonal antibody downstream purification is not only the most critically impacted process by increased antibody titer resulting from optimized mammalian cell culture expression systems, but also the most important purification step in determining overall process throughput, product quality, and economics. Advances in separation technology for capturing antibodies from complex feedstocks have been one focus of downstream purification process innovation for past 10 years. In this study, we evaluated new generation chromatography resins used in the antibody capture process including Protein A, cation exchange, and mixed mode chromatography to address the benefits and unique challenges posed by each chromatography approach. Our results demonstrate the benefit of improved binding capacity of new generation Protein A resins, address the concern of high concentration surge caused aggregation when using new generation cation exchange resins with over 100mg/mL binding capacity, and highlight the potential of multimodal cation exchange resins for capture process design. The new landscape of capture chromatography technologies provides options to achieve overall downstream purification outcome with high product quality and process efficiency. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. New data on electron-beam purification of wastewater

    NASA Astrophysics Data System (ADS)

    Pikaev, A. K.

    2002-11-01

    Recent environmental applications of radiation technology, developed in the author's laboratory, are presented in this paper. They are electron-beam and coagulation purification of molasses distillery slops from distillery-produced ethyl alcohol by fermentation of plant materials, electron-beam purification of wastewater from carboxylic acids (for example, formic acid) and removal of petroleum products (diesel fuel, motor oil and residual fuel oil) from water by γ-irradiation.

  2. Membrane processing technology in the food industry: food processing, wastewater treatment, and effects on physical, microbiological, organoleptic, and nutritional properties of foods.

    PubMed

    Kotsanopoulos, Konstantinos V; Arvanitoyannis, Ioannis S

    2015-01-01

    Membrane processing technology (MPT) is increasingly used nowadays in a wide range of applications (demineralization, desalination, stabilization, separation, deacidification, reduction of microbial load, purification, etc.) in food industries. The most frequently applied techniques are electrodialysis (ED), reverse osmosis (RO), nanofiltration (NF), ultrafiltration (UF), and microfiltration (MF). Several membrane characteristics, such as pore size, flow properties, and the applied hydraulic pressure mainly determine membranes' potential uses. In this review paper the basic membrane techniques, their potential applications in a large number of fields and products towards the food industry, the main advantages and disadvantages of these methods, fouling phenomena as well as their effects on the organoleptic, qualitative, and nutritional value of foods are synoptically described. Some representative examples of traditional and modern membrane applications both in tabular and figural form are also provided.

  3. Analysis of cavitation effect for water purifier using electrolysis

    NASA Astrophysics Data System (ADS)

    Shin, Dong Ho; Ko, Han Seo; Lee, Seung Ho

    2015-11-01

    Water is a limited and vital resource, so it should not be wasted by pollution. A development of new water purification technology is urgent nowadays since the original and biological treatments are not sufficient. The microbubble-aided method was investigated for removal of algal in this study since it overcomes demerits of the existing purification technologies. Thus, the cavitation effect in a venturi-type tube using the electrolysis was analyzed. Ruthenium-coated titanium plates were used as electrodes. Optimum electrode interval and applied power were determined for the electrolysis. Then, the optimized electrodes were installed in the venturi-type tube for generating cavitation. The cavitation effect could be enhanced without any byproduct by the bubbly flow induced by the electrolysis. The optimum mass flow rate and current were determined for the cavitation with the electrolysis. Finally, the visualization techniques were used to count the cell number of algal and microbubbles for the confirmation of the performance. As a result, the energy saving and high efficient water purifier was fabricated in this study. This work was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Korean government (MEST) (No. 2013R1A2A2A01068653).

  4. Multi-copy entanglement purification with practical spontaneous parametric down conversion sources

    NASA Astrophysics Data System (ADS)

    Zhang, Shuai-Shuai; Shu, Qi; Zhou, Lan; Sheng, Yu-Bo

    2017-06-01

    Entanglement purification is to distill the high quality entanglement from the low quality entanglement with local operations and classical communications. It is one of the key technologies in long-distance quantum communication. We discuss an entanglement purification protocol (EPP) with spontaneous parametric down conversion (SPDC) sources, in contrast to previous EPP with multi-copy mixed states, which requires ideal entanglement sources. We show that the SPDC source is not an obstacle for purification, but can benefit the fidelity of the purified mixed state. This EPP works for linear optics and is feasible in current experiment technology. Project supported by the National Natural Science Foundation of China (Grant Nos. 11474168 and 61401222), the Natural Science Foundation of Jiangsu Province, China (Grant No. BK20151502), the Qing Lan Project in Jiangsu Province, China, and a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions, China.

  5. [Complex technology for water and wastewater disinfection and its industrial realization in prototype unit].

    PubMed

    Arakcheev, E N; Brunman, V E; Brunman, M V; Konyashin, A V; Dyachenko, V A; Petkova, A P

    Usage of complex automated electrolysis unit for drinking water disinfection and wastewater oxidation and coagulation is scoped, its ecological and energy efficiency is shown. Properties of technological process of anolyte production using membrane electrolysis of brine for water disinfection in municipal pipelines and potassium ferrate production using electrochemical dissolution of iron anode in NaOH solution for usage in purification plants are listed. Construction of modules of industrial prototype for anolyte and ferrate production and applied aspects of automation of complex electrolysis unit are proved. Results of approbation of electrolytic potassium ferrate for drinking water disinfection and wastewater, rain water and environmental water oxidation and coagulation are shown.

  6. Method for Rapid Purification of Class IIa Bacteriocins and Comparison of Their Activities

    PubMed Central

    Guyonnet, D.; Fremaux, C.; Cenatiempo, Y.; Berjeaud, J. M.

    2000-01-01

    A three-step method was developed for the purification of mesentericin Y105 (60% yield) from the culture supernatant of Leuconostoc mesenteroides Y105. The same procedure was successfully applied to the purification of five other anti-Listeria bacteriocins identified by mass spectrometry. Specific activities of the purified bacteriocins were compared. PMID:10742275

  7. [Studies on technology optimization for extraction and purification of total flavones from root bark of Artocarpus styracifolius].

    PubMed

    Ren, Gang; Liu, Rong-hua; Shao, Feng; Huang, Hui-lian; Wen, Li-rong

    2010-08-01

    To study the technology optimization for extraction and purification of total flavones from root bark of Artocarpus styracifolius. The optimum extraction conditions were investigated by the contents of the total flavones, using orthogonal test; Static adsorption capacity and desorption rate were employed as examine items for the screening of optimum macroporous resin and optimum technology for the purification of total flavones with selected macroporous were also investigated. The optimum extraction conditions were as follows: using 60% alcohol of seven times than amounts of original material soaking 12 hours,extracting once with hot reflux method at 50 degrees C. HPD-500 type macroporous resin showed better adsorption and desorption property. The optimum purification conditions were as follows: the sample solution was prepared at the concentration of 50.0 mg/mL, subjected to HPD-500 type macroporous resin column chromatography with a load ratio of 22.0 mg total flavones per gram of resin. After standing for 1 hour, the column was eluted with 4 BV water before being eluted with 4 BV 80% alcohol. The purity of the product was 86.4%, which enhanced the content of total flavones by 533%. The optimum conditions for extraction and purification of total flavones from root bark of Artocarpus styractifolius are convenient and practical, and could be used as a reference for industrial production.

  8. A new method of auxiliary purification for motor vehicle exhaust.

    PubMed

    Li, Dingqi

    2018-07-01

    As a result of the limitations of current purification technologies, purification efficiency is relatively low, particularly during startup or in the case of other abnormal automobile exhaust. Therefore, a new method of auxiliary purification is proposed in this paper. The acidic solution of potassium permanganate can oxidize carbon monoxide, nitrogen oxides and sulfur dioxide at relatively high temperatures and the alkaline solution of potassium permanganate can selectively absorb nitrogen oxide and sulfur dioxide. Therefore, we carried out the experiment using a solution of potassium permanganate and sulfuric acid as well as a solution of sodium carbonate and potassium permanganate, which served as the reagents for the auxiliary purification. The results of the test showed that after auxiliary purification by the acidic solution of potassium permanganate and the alkaline solution of potassium permanganate, the concentrations of carbon monoxide, hydrocarbons, nitrogen oxides and solid particles in the emissions were considerably lower than the concentrations prior to purification. It is possible to reduce the motor vehicle exhaust by the auxiliary purification of the solutions.

  9. Tandem Affinity Purification of Protein Complexes from Eukaryotic Cells.

    PubMed

    Ma, Zheng; Fung, Victor; D'Orso, Iván

    2017-01-26

    The purification of active protein-protein and protein-nucleic acid complexes is crucial for the characterization of enzymatic activities and de novo identification of novel subunits and post-translational modifications. Bacterial systems allow for the expression and purification of a wide variety of single polypeptides and protein complexes. However, this system does not enable the purification of protein subunits that contain post-translational modifications (e.g., phosphorylation and acetylation), and the identification of novel regulatory subunits that are only present/expressed in the eukaryotic system. Here, we provide a detailed description of a novel, robust, and efficient tandem affinity purification (TAP) method using STREP- and FLAG-tagged proteins that facilitates the purification of protein complexes with transiently or stably expressed epitope-tagged proteins from eukaryotic cells. This protocol can be applied to characterize protein complex functionality, to discover post-translational modifications on complex subunits, and to identify novel regulatory complex components by mass spectrometry. Notably, this TAP method can be applied to study protein complexes formed by eukaryotic or pathogenic (viral and bacterial) components, thus yielding a wide array of downstream experimental opportunities. We propose that researchers working with protein complexes could utilize this approach in many different ways.

  10. EUV tools: hydrogen gas purification and recovery strategies

    NASA Astrophysics Data System (ADS)

    Landoni, Cristian; Succi, Marco; Applegarth, Chuck; Riddle Vogt, Sarah

    2015-03-01

    The technological challenges that have been overcome to make extreme ultraviolet lithography (EUV) a reality have been enormous1. This vacuum driven technology poses significant purity challenges for the gases employed for purging and cleaning the scanner EUV chamber and source. Hydrogen, nitrogen, argon and ultra-high purity compressed dry air (UHPCDA) are the most common gases utilized at the scanner and source level. Purity requirements are tighter than for previous technology node tools. In addition, specifically for hydrogen, EUV tool users are facing not only gas purity challenges but also the need for safe disposal of the hydrogen at the tool outlet. Recovery, reuse or recycling strategies could mitigate the disposal process and reduce the overall tool cost of operation. This paper will review the types of purification technologies that are currently available to generate high purity hydrogen suitable for EUV applications. Advantages and disadvantages of each purification technology will be presented. Guidelines on how to select the most appropriate technology for each application and experimental conditions will be presented. A discussion of the most common approaches utilized at the facility level to operate EUV tools along with possible hydrogen recovery strategies will also be reported.

  11. Current submarine atmosphere control technology.

    PubMed

    Mazurek, W

    1998-01-01

    Air purification in submarines was introduced towards the end of World War II and was limited to the use of soda lime for the removal of carbon dioxide and oxygen candles for the regeneration of oxygen. The next major advances came with the advent of nuclear-powered submarines. These included the development of regenerative and, sometimes, energy-intensive processes for comprehensive atmosphere revitalization. With the present development of conventional submarines using air-independent propulsion there is a requirement for air purification similar to that of the nuclear-powered submarines but it is constrained by limited power and space. Some progress has been made in the development of new technology and the adoption of air purification equipment used in the nuclear-powered submarines for this application.

  12. Intein-mediated one-step purification of Escherichia coli secreted human antibody fragments.

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Wu, Wan-Yi; Miller, Keith D.; Coolbaugh, Michael

    In this work, we apply self-cleaving affinity tag technology to several target proteins secreted into the Escherichia coli periplasm, including two with disulfide bonds. The target proteins were genetically fused to a self-cleaving chitin-binding domain intein tag for purification via a chitin agarose affinity resin. By attaching the intein-tagged fusion genes to the PelB secretion leader sequence, the tagged target proteins were secreted to the periplasmic space and could be recovered in active form by simple osmotic shock. After chitin-affinity purification, the target proteins were released from the chitin-binding domain tag via intein self-cleaving. This was induced by a smallmore » change in pH from 8.5 to 6.5 at room temperature, allowing direct elution of the cleaved target protein from the chitin affinity resin. The target proteins include the E. coli maltose-binding protein and b-lactamase enzyme, as well as two human antibody fragments that contain disulfide bonds. In all cases, the target proteins were purified with good activity and yield, without the need for refolding. Overall, this work demonstrates the compatibility of the DI-CM intein with the PelB secretion system in E. coli, greatly expanding its potential to more complex proteins.« less

  13. NASA - Johnson Space Center's New Capabilities for Air Purification

    NASA Technical Reports Server (NTRS)

    Graf, John

    2015-01-01

    NASA has some unique and challenging air purification problems that cannot be adequately met with COTS technology: 1) ammonia removal from air, 2) hydrazine removal from air, 3) CO conversion to CO2 in low temperature, high humidity environments. NASA has sponsored the development of new sorbents and new catalysts. These new sorbents and catalysts work better than COTS technology for our application. If attendees have a need for an effective ammonia sorbent, an effective hydrazine sorbent, or an effective CO conversion catalyst, we should learn to see if NASA sponsored technology development can help.

  14. Study of positive and negative plasma catalytic oxidation of ethylene.

    PubMed

    Van Wesenbeeck, K; Hauchecorne, B; Lenaerts, S

    2017-06-01

    The effect of introducing a photocatalytically active coating inside a plasma unit is investigated. This technique combines the advantages of high product selectivity from catalysis and the fast start-up from plasma technology. In this study, a preselected TiO 2 coating is applied on the collector electrode of a DC corona discharge unit as non-thermal plasma reactor, in order to study the oxidation of ethylene. For both positive and negative polarities an enhanced mineralization is observed while the formation of by-products drastically decreases. The plasma catalytic unit gave the best results when using negative polarity at a voltage of 15 kV. This shows the potential of plasma catalysis as indoor air purification technology.

  15. An effective method for thallium bromide purification and research on crystal properties

    NASA Astrophysics Data System (ADS)

    Zheng, Zhiping; Meng, Fang; Gong, Shuping; Quan, Lin; Wang, Jing; Zhou, Dongxiang

    2012-06-01

    Thallium bromide (TlBr) is a promising candidate for room-temperature X- and gamma-ray detectors in view of its excellent intrinsic features. However, material purity and crystal quality concerns still limit the use of TlBr crystals as detectors. In this work, a combination of hydrothermal recrystallization (HR) and vacuum distillation (VD) methods were applied to purify TlBr salts prior to crystal growth. Trace impurities at the ppb/ppm level were determined by inductively coupled plasma mass spectroscopy (ICP-MS). The results showed that the impurity concentrations of the TlBr salt decreased significantly after HR and VD purification, and high performance of the resultant TlBr crystal in areas such as electrical and optical properties was achieved. The combination of HR and VD methods could fabricate purer material, with an order of magnitude higher resistivity and better optical quality, than HR or VD method used separately. The possible technological considerations affecting the parameters of the crystals are investigated.

  16. Nanofluidic Lab-On-Chip Technology for DNA Identification

    DTIC Science & Technology

    2013-09-30

    samples Fluorescently labeled (FAM tag) DNA oligomers (10, 20, and 50 bases long) were purchased with standard desalting and additional HPLC purification...A.2 DNA samples: DNA oligomers (10, 20, 50 nt long) were purchased with standard desalting and additional HPLC purification for the 50 base

  17. Air/Water Purification

    NASA Technical Reports Server (NTRS)

    1992-01-01

    After 18 years of research into air/water pollution at Stennis Space Center, Dr. B. C. Wolverton formed his own company, Wolverton Environmental Services, Inc., to provide technology and consultation in air and water treatment. Common houseplants are used to absorb potentially harmful materials from bathrooms and kitchens. The plants are fertilized, air is purified, and wastewater is converted to clean water. More than 100 U.S. communities have adopted Wolverton's earlier water hyacinth and artificial marsh applications. Catfish farmers are currently evaluating the artificial marsh technology as a purification system.

  18. Purity and the dangers of regenerative medicine: regulatory innovation of human tissue-engineered technology.

    PubMed

    Faulkner, Alex; Kent, Julie; Geesink, Ingrid; FitzPatrick, David

    2006-11-01

    This paper examines the development of innovation in human tissue technologies as a form of regenerative medicine, firstly by applying 'pollution ideas' to contemporary trends in its risk regulation and to the processes of regulatory policy formation, and secondly by analysing the classificatory processes deployed in regulatory policy. The analysis draws upon data from fieldwork and documentary materials with a focus on the UK and EU (2002-05) and explores four arenas: governance and regulatory policy; commercialisation and the market; 'evidentiality' manifest in evidence-based policy; and publics' and technology users' values and ethics. The analysis suggests that there is a trend toward 'purification' across these arenas, both material and socio-political. A common process of partitioning is found in stakeholders' attempts to define a clear terrain, which the field of tissue-engineered technology might occupy. We conclude that pollution ideas and partitioning processes are useful in understanding regulatory ordering and innovation in the emerging technological zone of human tissue engineering.

  19. Extraction and purification of high added value compounds from by-products of the winemaking chain using alternative/nonconventional processes/technologies.

    PubMed

    Yammine, Sami; Brianceau, Sylène; Manteau, Sébastien; Turk, Mohammad; Ghidossi, Rémy; Vorobiev, Eugène; Mietton-Peuchot, Martine

    2018-05-24

    Grape byproducts are today considered as a cheap source of valuable compounds since existent technologies allow the recovery of target compounds and their recycling. The goal of the current article is to explore the different recovery stages used by both conventional and alternative techniques and processes. Alternative pre-treatments techniques reviewed are: ultrasounds, pulsed electric fields and high voltage discharges. In addition, nonconventional solvent extraction under high pressure, specifically, supercritical fluid extraction and subcritical water extraction are discussed. Finally alternative purification technologies, for example membrane processing were also examined. The intent is to describe the mechanisms involved by these alternative technologies and to summarize the work done on the improvement of the extraction process of phenolic compounds from winery by-products. With a focus on the developmental stage of each technology, highlighting the research need and challenges to be overcome for an industrial implementation of these unitary operations in the overall extraction process. A critical comparison of conventional and alternative techniques will be reviewed for ethe pre-treatment of raw material, the diffusion of polyphenols and the purification of these high added value compounds. This review intends to give the reader some key answers (costs, advantages, drawbacks) to help in the choice of alternative technologies for extraction purposes.

  20. Comparison of a novel TiO₂/diatomite composite and pure TiO₂ for the purification of phosvitin phosphopeptides.

    PubMed

    Zhang, Yang; Li, Junhua; Niu, Fuge; Sun, Jun; Dou, Yuan; Liu, Yuntao; Su, Yujie; Zhou, Bei; Xu, Qinqin; Yang, Yanjun

    2014-06-01

    A novel TiO2/diatomite composite (TD) was prepared and then characterized by scanning electron microscope (SEM) and Fourier Transform Infrared (FTIR). The results of SEM showed that after modification, the porous surface of diatomite was covered with TiO2. Both diatomite and TD had clear disc-shaped structures with average grain diameters of around 25 μm. Then TD and pure TiO2 were applied in the purification of phosvitin phosphopeptides (PPPs) from the digest of egg yolk protein, and a comparative study of adsorption properties of PPPs on TD and TiO2 was performed. In the study of adsorption kinetics, the adsorption equilibrium of PPPs on TD and TiO2 fitted well with the Langmuir model, and the time needed to reach adsorption equilibrium were both around 10 min. The maximum dynamic adsorption capacity of TD (8.15 mg/g) was higher than that of TiO2 (4.96 mg/g). The results of repeated use showed that TD and TiO2 were very stable after being subjected to ten repeated adsorption-elution cycles, and TD could easily be separated from aqueous solution by filtration. On the other hand, the present synthetic technology of TD was very simple, cost-effective, organic solvent-free and available for large-scale preparation. Thus, this separation method not only brings great advantages in the purification of PPPs from egg yolk protein but also provides a promising purification material for the enrichment of phosphopeptides in proteomic researches. Copyright © 2014 Elsevier B.V. All rights reserved.

  1. TiO2-Based Advanced Oxidation Nanotechnologies For Water Purification And Reuse

    EPA Science Inventory

    TiO2 photocatalysis, one of the UV-based advanced oxidation technologies (AOTs) and nanotechnologies (AONs), has attracted great attention for the development of efficient water treatment and purification systems due to the effectiveness of TiO2 to generate ...

  2. Evaluation of the effectiveness of semen storage and sperm purification methods for spermatozoa transcript profiling.

    PubMed

    Mao, Shihong; Goodrich, Robert J; Hauser, Russ; Schrader, Steven M; Chen, Zhen; Krawetz, Stephen A

    2013-10-01

    Different semen storage and sperm purification methods may affect the integrity of isolated spermatozoal RNA. RNA-Seq was applied to determine whether semen storage methods (pelleted vs. liquefied) and somatic cell lysis buffer (SCLB) vs. PureSperm (PS) purification methods affect the quantity and quality of sperm RNA. The results indicate that the method of semen storage does not markedly impact RNA profiling whereas the choice of purification can yield significant differences. RNA-Seq showed that the majority of mitochondrial and mid-piece associated transcripts were lost after SCLB purification, which indicated that the mid-piece of spermatozoa may have been compromised. In addition, the number of stable transcript pairs from SCLB-samples was less than that from the PS samples. This study supports the view that PS purification better maintains the integrity of spermatozoal RNAs.

  3. Two-step purification method of vitellogenin from three teleost fish species: rainbow trout (Oncorhynchus mykiss), gudgeon (Gobio gobio) and chub (Leuciscus cephalus).

    PubMed

    Brion, F; Rogerieux, F; Noury, P; Migeon, B; Flammarion, P; Thybaud, E; Porcher, J M

    2000-01-14

    A two-step purification protocol was developed to purify rainbow trout (Oncorhynchus mykiss) vitellogenin (Vtg) and was successfully applied to Vtg of chub (Leuciscus cephalus) and gudgeon (Gobio gobio). Capture and intermediate purification were performed by anion-exchange chromatography on a Resource Q column and a polishing step was performed by gel permeation chromatography on Superdex 200 column. This method is a rapid two-step purification procedure that gave a pure solution of Vtg as assessed by silver staining electrophoresis and immunochemical characterisation.

  4. Production of alpha-amylase from Aspergillus oryzae for several industrial applications in a single step.

    PubMed

    Porfirif, María C; Milatich, Esteban J; Farruggia, Beatriz M; Romanini, Diana

    2016-06-01

    A one-step method as a strategy of alpha-amylase concentration and purification was developed in this work. This methodology requires the use of a very low concentration of biodegradable polyelectrolyte (Eudragit(®) E-PO) and represents a low cost, fast, easy to scale up and non-polluting technology. Besides, this methodology allows recycling the polymer after precipitation. The formation of reversible soluble/insoluble complexes between alpha-amylase and the polymer Eudragit(®) E-PO was studied, and their precipitation in selected conditions was applied with bioseparation purposes. Turbidimetric assays allowed to determine the pH range where the complexes are insoluble (4.50-7.00); pH 5.50 yielded the highest turbidity of the system. The presence of NaCl (0.05M) in the medium totally dissociates the protein-polymer complexes. When the adequate concentration of polymer was added under these conditions to a liquid culture of Aspergillus oryzae, purification factors of alpha-amylase up to 7.43 and recoveries of 88% were obtained in a simple step without previous clarification. These results demonstrate that this methodology is suitable for the concentration and production of alpha-amylase from this source and could be applied at the beginning of downstream processing. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. [Extraction and purification technologies of total flavonoids from Aconitum tanguticum].

    PubMed

    Li, Yan-Rong; Yan, Li-Xin; Feng, Wei-Hong; Li, Chun; Wang, Zhi-Min

    2014-04-01

    To optimize the extraction and purification technologies of total flavonoids from Aconitum tanguticum whole plant. With the content of total flavonoids as index, the optimum extraction conditions for the concentration, volume of alcohol, extracting time and times were selected by orthogonal optimized; Comparing the adsorption quantity (mg/g) and resolution (%), four kinds of macroporous adsorption resins including D101, AB-8, X-5 and XAD-16 were investigated for the enrichment ability of total flavonoids from Aconitum tanguticum; Concentration and pH value of sample, sampling amount, elution solvent and loading and elution velocity for the optimum adsorption resin were determined. The content of total flavonoids in Aconitum tanguticum was about 4.39%; The optimum extraction technique was 70% alcohol reflux extraction for three times,each time for one hour, the ratio of material and liquid was 1:10 (w/v); The optimum purification technology was: using XAD-16 macroporous resin, the initial concentration of total flavonoids of Aconitum tanguticum was 8 mg/mL, the sampling amount was 112 mg/g dry resin, the pH value was 5, the loading velocity was 3 mL/min, the elution solvent was 70% ethanol and the elution velocity was 5 mL/min. Under the optimum conditions, the average content of total flavonoids was raised from 4.39% to 46.19%. The optimum extraction and purification technologies for total flavonoids of Aconitum tanguticum were suitable for industrial production for its simplicity and responsibility.

  6. Study on the Key Technology of High Purity Strontium Titanate Powder Synthesized from Oxalic Acid Co-sediment Precipitation

    NASA Astrophysics Data System (ADS)

    Bi, Xiaoguo; Dong, Yingnan; Li, Yingjie; Niu, Wei; Tang, Jian; Ding, Shuang; Li, Meiyang

    2017-09-01

    Oxalate coprecipitation is applied in this paper, high purity titanium tetrachloride, and after the purification of strontium chloride, match with a certain concentration of solution, oxalate and strontium chloride and titanium tetrachloride in 1.005:1.000 make strontium titanium mixture ratio, slowly under 60°C to join in oxalic acid solution, aging around 4 h, get oxygen titanium strontium oxalate (SrTiO(C2O4)2 • 4H2 ) precipitation, after washing, drying and other process made oxygen titanium strontium oxalate powder.

  7. Automated high-throughput purification of genomic DNA from plant leaf or seed using MagneSil paramagnetic particles

    NASA Astrophysics Data System (ADS)

    Bitner, Rex M.; Koller, Susan C.

    2004-06-01

    Three different methods of automated high throughput purification of genomic DNA from plant materials processed in 96 well plates are described. One method uses MagneSil paramagnetic particles to purify DNA present in single leaf punch samples or small seed samples, using 320ul capacity 96 well plates which minimizes reagent and plate costs. A second method uses 2.2 ml and 1.2 ml capacity plates and allows the purification of larger amounts of DNA from 5-6 punches of materials or larger amounts of seeds. The third method uses the MagneSil ONE purification system to purify a fixed amount of DNA, thus simplifying the processing of downstream applications by normalizing the amounts of DNA so they do not require quantitation. Protocols for the purification of a fixed yield of DNA, e.g. 1 ug, from plant leaf or seed samples using MagneSil paramagnetic particles and a Beckman-Coulter BioMek FX robot are described. DNA from all three methods is suitable for applications such as PCR, RAPD, STR, READIT SNP analysis, and multiplexed PCR systems. The MagneSil ONE system is also suitable for use with SNP detection systems such as Third Wave Technology"s Invader methods.

  8. Environmental Technology Verification Report - NATCO Group, Inc. – Paques THIOPAQ Gas Purification Technology

    EPA Science Inventory

    The U.S. Environmental Protection Agency (EPA) has created the Environmental Technology Verification (ETV) program to facilitate the deployment of innovative or improved environmental technologies through performance verification and dissemination of information. The goal of the ...

  9. Sophisticated Cloning, Fermentation, and Purification Technologies for an Enhanced Therapeutic Protein Production: A Review

    PubMed Central

    Gupta, Sanjeev K.; Shukla, Pratyoosh

    2017-01-01

    The protein productions strategies are crucial towards the development of application based research and elucidating the novel purification strategies for industrial production. Currently, there are few innovative avenues are studies for cloning, upstream, and purification through efficient bioprocess development. Such strategies are beneficial for industries as well as proven to be vital for effectual therapeutic protein development. Though, these techniques are well documented, but, there is scope of addition to current knowledge with novel and new approaches and it will pave new avenues in production of recombinant microbial and non-microbial proteins including secondary metabolites. In this review, we have focussed on the recent development in clone selection, various modern fermentation and purification technologies and future directions in these emerging areas. Moreover, we have also highlighted notable perspectives and challenges involved in the bioengineering of such proteins, including quality by design, gene editing and pioneering ideas. The biopharmaceutical industries continue to shift towards more flexible, automated platforms and economical product development, which in turn can help in developing the cost effective processes and affordable drug development for a large community. PMID:28725194

  10. Production of recombinant protein G through high-density fermentation of engineered bacteria as well as purification.

    PubMed

    Zhang, Hu-Cheng; Yang, Jun; Yang, Guo-Wei; Wang, Xiao-Jie; Fan, Hai-Tao

    2015-08-01

    Recombinant Streptococcus Protein G (PG) is a cell wall protein, which, when combined with mammal immunoglobulin, is used in separating antibody technology. High-density fermentation technologies using an engineered recombinant PG-producing bacteria as well as PG separation and purification technologies have a direct impact on the availability and application of PG. Through primary and secondary seed cultivation, a recombinant E. coli strain was subjected to high-density fermentation with controlled feed supplement concentration under stimulation with isopropyl β-D-1-thiogalactopyranoside. The present study investigated the effect of factors including inoculum size, oxygen levels, pH and the cultivating method on the fermentation process, as well as the effect of the separation and purification technologies, including ultrasonication, nickel column affinity chromatography, Sephadex G-25 gel filtration chromatography and diethylaminoethanol-sepharose fast flow ion exchange chromatography on the yield and purity of PG. The efficiency of extraction was detected using SDS-PAGE. High-density fermentation yielded 80-150 g/l of bacteria and 1 g PG was obtained from one liter broth. The present study delivered a highly efficient novel method via which PG can be obtained at a high concentration and a purity >95%.

  11. Establishment of cell surface engineering and its development.

    PubMed

    Ueda, Mitsuyoshi

    2016-07-01

    Cell surface display of proteins/peptides has been established based on mechanisms of localizing proteins to the cell surface. In contrast to conventional intracellular and extracellular (secretion) expression systems, this method, generally called an arming technology, is particularly effective when using yeasts as a host, because the control of protein folding that is often required for the preparation of proteins can be natural. This technology can be employed for basic and applied research purposes. In this review, I describe various strategies for the construction of engineered yeasts and provide an outline of the diverse applications of this technology to industrial processes such as the production of biofuels and chemicals, as well as bioremediation and health-related processes. Furthermore, this technology is suitable for novel protein engineering and directed evolution through high-throughput screening, because proteins/peptides displayed on the cell surface can be directly analyzed using intact cells without concentration and purification. Functional proteins/peptides with improved or novel functions can be created using this beneficial, powerful, and promising technique.

  12. A CRISPR/Cas9 and Cre/Lox system-based express vaccine development strategy against re-emerging Pseudorabies virus.

    PubMed

    Liang, Xun; Sun, Leqiang; Yu, Teng; Pan, Yongfei; Wang, Dongdong; Hu, Xueying; Fu, Zhenfang; He, Qigai; Cao, Gang

    2016-01-18

    Virus evolves rapidly to escape vaccine-induced immunity, posing a desperate demand for efficient vaccine development biotechnologies. Here we present an express vaccine development strategy based on CRISPR/Cas9 and Cre/Lox system against re-emerging Pseudorabies virus, which caused the recent devastating swine pseudorabies outbreak in China. By CRISPR/Cas9 system, the virulent genes of the newly isolated strain were simultaneously substituted by marker genes, which were subsequently excised using Cre/Lox system for vaccine safety concern. Notably, single cell FACS technology was applied to further promote virus purification efficiency. The combination of these state-of-art technologies greatly accelerated vaccine development. Finally, vaccination and challenge experiments proved this vaccine candidate's protective efficacy in pigs and the promise to control current pseudorabies outbreak. This is, to our knowledge, the first successful vaccine development based on gene edit technologies, demonstrating these technologies leap from laboratory to industry. It may pave the way for future express antiviral vaccine development.

  13. A CRISPR/Cas9 and Cre/Lox system-based express vaccine development strategy against re-emerging Pseudorabies virus

    PubMed Central

    Liang, Xun; Sun, Leqiang; Yu, Teng; Pan, Yongfei; Wang, Dongdong; Hu, Xueying; Fu, Zhenfang; He, Qigai; Cao, Gang

    2016-01-01

    Virus evolves rapidly to escape vaccine-induced immunity, posing a desperate demand for efficient vaccine development biotechnologies. Here we present an express vaccine development strategy based on CRISPR/Cas9 and Cre/Lox system against re-emerging Pseudorabies virus, which caused the recent devastating swine pseudorabies outbreak in China. By CRISPR/Cas9 system, the virulent genes of the newly isolated strain were simultaneously substituted by marker genes, which were subsequently excised using Cre/Lox system for vaccine safety concern. Notably, single cell FACS technology was applied to further promote virus purification efficiency. The combination of these state-of-art technologies greatly accelerated vaccine development. Finally, vaccination and challenge experiments proved this vaccine candidate’s protective efficacy in pigs and the promise to control current pseudorabies outbreak. This is, to our knowledge, the first successful vaccine development based on gene edit technologies, demonstrating these technologies leap from laboratory to industry. It may pave the way for future express antiviral vaccine development. PMID:26777545

  14. ELISA AND SOL-GEL BASED IMMUNOAFFINITY PURIFICATION OF THE PYRETHROID BIOALLETHRIN IN FOOD AND ENVIRONMENTAL SAMPLES

    EPA Science Inventory

    The peer-reviewed article describes the development of a new sol-gel based immunoaffinity purification procedure and an immunoassay for the pyrethroid bioallethrin. The immunoaffinity chromatography procedure was applied to food samples providing an efficient cleanup prior to im...

  15. Purification of crime scene DNA extracts using centrifugal filter devices

    PubMed Central

    2013-01-01

    Background The success of forensic DNA analysis is limited by the size, quality and purity of biological evidence found at crime scenes. Sample impurities can inhibit PCR, resulting in partial or negative DNA profiles. Various DNA purification methods are applied to remove impurities, for example, employing centrifugal filter devices. However, irrespective of method, DNA purification leads to DNA loss. Here we evaluate the filter devices Amicon Ultra 30 K and Microsep 30 K with respect to recovery rate and general performance for various types of PCR-inhibitory crime scene samples. Methods Recovery rates for DNA purification using Amicon Ultra 30 K and Microsep 30 K were gathered using quantitative PCR. Mock crime scene DNA extracts were analyzed using quantitative PCR and short tandem repeat (STR) profiling to test the general performance and inhibitor-removal properties of the two filter devices. Additionally, the outcome of long-term routine casework DNA analysis applying each of the devices was evaluated. Results Applying Microsep 30 K, 14 to 32% of the input DNA was recovered, whereas Amicon Ultra 30 K retained 62 to 70% of the DNA. The improved purity following filter purification counteracted some of this DNA loss, leading to slightly increased electropherogram peak heights for blood on denim (Amicon Ultra 30 K and Microsep 30 K) and saliva on envelope (Amicon Ultra 30 K). Comparing Amicon Ultra 30 K and Microsep 30 K for purification of DNA extracts from mock crime scene samples, the former generated significantly higher peak heights for rape case samples (P-values <0.01) and for hairs (P-values <0.036). In long-term routine use of the two filter devices, DNA extracts purified with Amicon Ultra 30 K were considerably less PCR-inhibitory in Quantifiler Human qPCR analysis compared to Microsep 30 K. Conclusions Amicon Ultra 30 K performed better than Microsep 30 K due to higher DNA recovery and more efficient removal of PCR-inhibitory substances. The different performances of the filter devices are likely caused by the quality of the filters and plastic wares, for example, their DNA binding properties. DNA purification using centrifugal filter devices can be necessary for successful DNA profiling of impure crime scene samples and for consistency between different PCR-based analysis systems, such as quantification and STR analysis. In order to maximize the possibility to obtain complete STR DNA profiles and to create an efficient workflow, the level of DNA purification applied should be correlated to the inhibitor-tolerance of the STR analysis system used. PMID:23618387

  16. Effective removal of co-purified inhibitors from extracted DNA samples using synchronous coefficient of drag alteration (SCODA) technology.

    PubMed

    Schmedes, Sarah; Marshall, Pamela; King, Jonathan L; Budowle, Bruce

    2013-07-01

    Various types of biological samples present challenges for extraction of DNA suitable for subsequent molecular analyses. Commonly used extraction methods, such as silica membrane columns and phenol-chloroform, while highly successful may still fail to provide a sufficiently pure DNA extract with some samples. Synchronous coefficient of drag alteration (SCODA), implemented in Boreal Genomics' Aurora Nucleic Acid Extraction System (Boreal Genomics, Vancouver, BC), is a new technology that offers the potential to remove inhibitors effectively while simultaneously concentrating DNA. In this initial study, SCODA was tested for its ability to remove various concentrations of forensically and medically relevant polymerase chain reaction (PCR) inhibitors naturally found in tissue, hair, blood, plant, and soil samples. SCODA was used to purify and concentrate DNA from intentionally contaminated DNA samples containing known concentrations of hematin, humic acid, melanin, and tannic acid. The internal positive control (IPC) provided in the Quantifiler™ Human DNA Quantification Kit (Life Technologies, Foster City, CA) and short tandem repeat (STR) profiling (AmpFℓSTR® Identifiler® Plus PCR Amplification Kit; Life Technologies, Foster City, CA) were used to measure inhibition effects and hence purification. SCODA methodology yielded overall higher efficiency of purification of highly contaminated samples compared with the QIAquick® PCR Purification Kit (Qiagen, Valencia, CA). SCODA-purified DNA yielded no cycle shift of the IPC for each sample and yielded greater allele percentage recovery and relative fluorescence unit values compared with the QIAquick® purification method. The Aurora provided an automated, minimal-step approach to successfully remove inhibitors and concentrate DNA from challenged samples.

  17. Bridging the Gap: Use of Spaceflight Technologies for Earth-Based Problems

    NASA Technical Reports Server (NTRS)

    Brinley, Alaina; Vidlak, Carissa; Davis, Jeffrey R.

    2012-01-01

    Spaceflight is colloquially deemed, the final frontier, or the last area which humans have not yet explored in great depth. While this is true, there are still many regions on Earth that remain isolated from the urban, socially and electronically connected world. Because travelling to space requires a great deal of foresight, engineers are required to think creatively in order to invent technologies that are durable enough to withstand the rigors of the unique and often treacherous environment of outer space. The innovations that are a result of spaceflight designs can often be applied to life on Earth, particularly in the rural, isolated communities found throughout the world. The NASA Human Health and Performance Center (NHHPC) is a collaborative, virtual forum that connects businesses, non-profit organizations, academia, and government agencies to allow for better distribution of ideas and technology between these entities (http://www.nasa.gov/offices/NHHPC). There are many technologies that have been developed for spaceflight that can be readily applied to rural communities on Earth. For example, water filtration systems designed for spaceflight must be robust and easily repaired; therefore, a system with these qualifications may be used in rural areas on Earth. This particular initiative seeks to connect established, non-profit organizations working in isolated communities throughout the world with NASA technologies devised for spaceflight. These technologies could include water purification systems, solar power generators, or telemedicine techniques. Applying innovative, spaceflight technologies to isolated communities on Earth provides greater benefits from the same research dollars, thus fulfilling the Space Life Science motto at Johnson Space Center: Exploring Space and Enhancing Life. This paper will discuss this NHHPC global outreach initiative and give examples based on the recent work of the organization.

  18. Purification of nanogram-range immunoprecipitated DNA in ChIP-seq application.

    PubMed

    Zhong, Jian; Ye, Zhenqing; Lenz, Samuel W; Clark, Chad R; Bharucha, Adil; Farrugia, Gianrico; Robertson, Keith D; Zhang, Zhiguo; Ordog, Tamas; Lee, Jeong-Heon

    2017-12-21

    Chromatin immunoprecipitation-sequencing (ChIP-seq) is a widely used epigenetic approach for investigating genome-wide protein-DNA interactions in cells and tissues. The approach has been relatively well established but several key steps still require further improvement. As a part of the procedure, immnoprecipitated DNA must undergo purification and library preparation for subsequent high-throughput sequencing. Current ChIP protocols typically yield nanogram quantities of immunoprecipitated DNA mainly depending on the target of interest and starting chromatin input amount. However, little information exists on the performance of reagents used for the purification of such minute amounts of immunoprecipitated DNA in ChIP elution buffer and their effects on ChIP-seq data. Here, we compared DNA recovery, library preparation efficiency, and ChIP-seq results obtained with several commercial DNA purification reagents applied to 1 ng ChIP DNA and also investigated the impact of conditions under which ChIP DNA is stored. We compared DNA recovery of ten commercial DNA purification reagents and phenol/chloroform extraction from 1 to 50 ng of immunopreciptated DNA in ChIP elution buffer. The recovery yield was significantly different with 1 ng of DNA while similar in higher DNA amounts. We also observed that the low nanogram range of purified DNA is prone to loss during storage depending on the type of polypropylene tube used. The immunoprecipitated DNA equivalent to 1 ng of purified DNA was subject to DNA purification and library preparation to evaluate the performance of four better performing purification reagents in ChIP-seq applications. Quantification of library DNAs indicated the selected purification kits have a negligible impact on the efficiency of library preparation. The resulting ChIP-seq data were comparable with the dataset generated by ENCODE consortium and were highly correlated between the data from different purification reagents. This study provides comparative data on commercial DNA purification reagents applied to nanogram-range immunopreciptated ChIP DNA and evidence for the importance of storage conditions of low nanogram-range purified DNA. We verified consistent high performance of a subset of the tested reagents. These results will facilitate the improvement of ChIP-seq methodology for low-input applications.

  19. [Alcohol-purification technology and its particle sedimentation process in manufactory of Fufang Kushen injection].

    PubMed

    Liu, Xiaoqian; Tong, Yan; Wang, Jinyu; Wang, Ruizhen; Zhang, Yanxia; Wang, Zhimin

    2011-11-01

    Fufang Kushen injection was selected as the model drug, to optimize its alcohol-purification process and understand the characteristics of particle sedimentation process, and to investigate the feasibility of using process analytical technology (PAT) on traditional Chinese medicine (TCM) manufacturing. Total alkaloids (calculated by matrine, oxymatrine, sophoridine and oxysophoridine) and macrozamin were selected as quality evaluation markers to optimize the process of Fufang Kushen injection purification with alcohol. Process parameters of particulate formed in the alcohol-purification, such as the number, density and sedimentation velocity, were also determined to define the sedimentation time and well understand the process. The purification process was optimized as that alcohol is added to the concentrated extract solution (drug material) to certain concentration for 2 times and deposited the alcohol-solution containing drug-material to sediment for some time, i.e. 60% alcohol deposited for 36 hours, filter and then 80% -90% alcohol deposited for 6 hours in turn. The content of total alkaloids was decreased a little during the depositing process. The average settling time of particles with the diameters of 10, 25 microm were 157.7, 25.2 h in the first alcohol-purified process, and 84.2, 13.5 h in the second alcohol-purified process, respectively. The optimized alcohol-purification process remains the marker compositions better and compared with the initial process, it's time saving and much economy. The manufacturing quality of TCM-injection can be controlled by process. PAT pattern must be designed under the well understanding of process of TCM production.

  20. Vacuum isostatic micro/macro molding of PTFE materials for laser beam shaping in environmental applications: large scale UV laser water purification

    NASA Astrophysics Data System (ADS)

    Lizotte, Todd; Ohar, Orest

    2009-08-01

    Accessibility to fresh clean water has determined the location and survival of civilizations throughout the ages [1]. The tangible economic value of water is demonstrated by industry's need for water in fields such as semiconductor, food and pharmaceutical manufacturing. Economic stability for all sectors of industry depends on access to reliable volumes of good quality water. As can be seen on television a nation's economy is seriously affected by water shortages through drought or mismanagement and as such those water resources must therefore be managed both for the public interest and the economic future. For over 50 years ultraviolet water purification has been the mainstay technology for water treatment, killing potential microbiological agents in water for leisure activities such as swimming pools to large scale waste water treatment facilities where the UV light photo-oxidizes various pollutants and contaminants. Well tailored to the task, UV provides a cost effective way to reduce the use of chemicals in sanitization and anti-biological applications. Predominantly based on low pressure Hg UV discharge lamps, the system is plagued with lifetime issues (~1 year normal operation), the last ten years has shown that the technology continues to advance and larger scale systems are turning to more advanced lamp designs and evaluating solidstate UV light sources and more powerful laser sources. One of the issues facing the treatment of water with UV lasers is an appropriate means of delivering laser light efficiently over larger volumes or cross sections of water. This paper examines the potential advantages of laser beam shaping components made from isostatically micro molding microstructured PTFE materials for integration into large scale water purification and sterilization systems, for both lamps and laser sources. Applying a unique patented fabrication method engineers can form micro and macro scale diffractive, holographic and faceted reflective structures into fused and semi-fused PTFE materials and compounds for use in UV Reactors. The materials unique attributes provide an unusual but effective hybrid element, by combining Lambertian diffusion and spectral reflective attributes. This paper will provide examples of the applications where this technology could be applied and typical constructions. An overview of UV sources commonly used in water treatment, including high power UV lasers and solid state UV light sources will be discussed. The paper will summarize how beam shaping elements produced in PTFE materials would provide further benefits to the emerging water disinfection or treatment market.

  1. Rationale for constructing waste-disposal plants at existing enterprises

    NASA Astrophysics Data System (ADS)

    Strelkov, Alexander; Teplykh, Svetlana; Gorshkalev, Pavel; Proshina, Elizaveta

    2017-10-01

    The Federal State Statistics Service of the Republic of Tatarstan collected data on registered organizations involved in the fabrication and dyeing of fur. This paper describes wastewater characteristic of an existing enterprise, LLC “Melita”. This enterprise is a factory of a complete technology cycle, with the process starting from fur manufacture and design to implementation. Maximum capacity of the factory is 1800 skins per day, (excluding fur), and its average productivity is 1000 skins per day. Thorough examination of possible methods and schemes for wastewater from fur production purification showed that it is most reasonable to use technology schemes which included the structures of mechanical, physico-chemical and biological purification. As a result, the study provided a new technological scheme for industrial wastewater purification. This scheme offers using chloride barium and sodium hydrocarbons complex as reagents. For LLC “Melita”, the wastewater absorbing pond is the Volga River. Water quality indicators are taken according to the data of the FGBU “Hydrometeorology and Environmental Monitoring Office”. The research also calculates allowable discharge rates and environmental charges in the city sewer networks and ponds.

  2. Recent advances in cellulose and chitosan based membranes for water purification: A concise review.

    PubMed

    Thakur, Vijay Kumar; Voicu, Stefan Ioan

    2016-08-01

    Recently membrane technology has emerged as a new promising and pervasive technology due to its innate advantages over traditional technologies such as adsorption, distillation and extraction. In this article, some of the recent advances in developing polymeric composite membrane materials for water purification from natural polysaccharide based polymers namely cellulose derivatives and chitosan are concisely reviewed. The impact of human social, demographic and industrial evolution along with expansion through environment has significantly affected the quality of water by pollution with large quantities of pesticides, minerals, drugs or other residues. At the forefront of decontamination and purification techniques, we found the membrane materials from polymers as a potential alternative. In an attempt to reduce the number of technical polymers widely used in the preparation of membranes, many researchers have reported new solutions for desalination or retention of organic yeasts, based on bio renewable polymers like cellulose derivatives and chitosan. These realizations are presented and discussed in terms of the most important parameters of membrane separation especially water flux and retention in this article. Published by Elsevier Ltd.

  3. Halophilic viruses with varying biochemical and biophysical properties are amenable to purification with asymmetrical flow field-flow fractionation.

    PubMed

    Eskelin, Katri; Lampi, Mirka; Meier, Florian; Moldenhauer, Evelin; Bamford, Dennis H; Oksanen, Hanna M

    2017-11-01

    Viruses come in various shapes and sizes, and a number of viruses originate from extremities, e.g. high salinity or elevated temperature. One challenge for studying extreme viruses is to find efficient purification conditions where viruses maintain their infectivity. Asymmetrical flow field-flow fractionation (AF4) is a gentle native chromatography-like technique for size-based separation. It does not have solid stationary phase and the mobile phase composition is readily adjustable according to the sample needs. Due to the high separation power of specimens up to 50 µm, AF4 is suitable for virus purification. Here, we applied AF4 for extremophilic viruses representing four morphotypes: lemon-shaped, tailed and tailless icosahedral, as well as pleomorphic enveloped. AF4 was applied to input samples of different purity: crude supernatants of infected cultures, polyethylene glycol-precipitated viruses and viruses purified by ultracentrifugation. All four virus morphotypes were successfully purified by AF4. AF4 purification of culture supernatants or polyethylene glycol-precipitated viruses yielded high recoveries, and the purities were comparable to those obtained by the multistep ultracentrifugation purification methods. In addition, we also demonstrate that AF4 is a rapid monitoring tool for virus production in slowly growing host cells living in extreme conditions.

  4. [Investigation on the process of sapindus saponin purified with macroporous adsorption resin and screening of its bacteriostasis].

    PubMed

    Fu, Yong; Lei, Peng; Han, Yu-mei; Yan, Dan

    2010-02-01

    To study the technological parameters of the purification process of saponins with macroporous adsorption resin. The adsorptive characteristics and elutive parameters of the process were studied by taking the elutive and purified ratio of saponins as markers. Bacteriostasis activity of each parts eluted was evaluated by the mean of cup-plate method. 13.6 mL of the extraction of sapindus saponin (crude drugs 0.01 g/mL) was purified with a column of macroporous adsorption resin (phi15 mm x H90 mm, dry weight 2.5 g) and washed with 3BV of distilled water, then eluted with 3BV of 30% ethanol and 3BV of 70% ethanol, most of saponins were collected in the 70% ethanol. With macroporous adsorption resin adsorbing and purifying, the elutive ratio of saponins was 93.8% and the purity reached 250.1%. So this process of applying macroporous adsorption resin to adsorb and purify saponins is feasible, and supplies reference to the purification of other types of saponin.

  5. Scalable Electrophysiological Investigation of iPS Cell-Derived Cardiomyocytes Obtained by a Lentiviral Purification Strategy.

    PubMed

    Friedrichs, Stephanie; Malan, Daniela; Voss, Yvonne; Sasse, Philipp

    2015-01-08

    Disease-specific induced pluripotent stem (iPS) cells can be generated from patients and differentiated into functional cardiomyocytes for characterization of the disease and for drug screening. In order to obtain pure cardiomyocytes for automated electrophysiological investigation, we here report a novel non-clonal purification strategy by using lentiviral gene transfer of a puromycin resistance gene under the control of a cardiac-specific promoter. We have applied this method to our previous reported wild-type and long QT syndrome 3 (LQTS 3)-specific mouse iPS cells and obtained a pure cardiomyocyte population. These cells were investigated by action potential analysis with manual and automatic planar patch clamp technologies, as well as by recording extracellular field potentials using a microelectrode array system. Action potentials and field potentials showed the characteristic prolongation at low heart rates in LQTS 3-specific, but not in wild-type iPS cell-derived cardiomyocytes. Hence, LQTS 3-specific cardiomyocytes can be purified from iPS cells with a lentiviral strategy, maintain the hallmarks of the LQTS 3 disease and can be used for automated electrophysiological characterization and drug screening.

  6. Purification of silicon for photovoltaic applications

    NASA Astrophysics Data System (ADS)

    Delannoy, Yves

    2012-12-01

    Solar grade silicon, as a starting material for crystallization to produce solar cells, is discussed here in terms of impurities whose maximum content is estimated from recent literature and conferences. A review of the production routes for each category of solar-grade silicon (undoped, compensated or heavily compensated) is proposed with emphasis on the metallurgical route. Some recent results are proposed concerning segregation, showing that directional solidification systems can be used for solidification even at high solidification rate (15 cm/h). Results on inductive plasma purification, where boron is evacuated as HBO in a gas phase blown from an inductive plasma torch, are shown to apply as well to arc plasmas and purification by moist gas. Special attention is paid to the history of impurities in the purification processes, showing that impure auxiliary phases (silicon tetrachloride, slag, aluminum, etc.) often need their own purification process to enable their recycling, which has to be considered to evaluate the cost (financial, energetic and environmental) of the purification route.

  7. Effects of indoor air purification by an air cleaning system (Koala technology) on semen parameters in male factor infertility: results of a pilot study.

    PubMed

    Paradisi, R; Vanella, S; Barzanti, R; Cani, C; Battaglia, C; Seracchioli, R; Venturoli, S

    2009-06-01

    A number of studies indicated a clear decline in semen quality in the past 30-50 years and there is accumulating evidence that this decline might result from exposure to high levels of air pollution. To examine the impact of environment on male reproductive ability, we undertook for the first time a pilot study on semen quality of infertile men exposed to purification of indoor air. Ten subjects with a history of unexplained male infertility and poor semen quality were exposed for at least 1 year to a cleaning indoor air system (Koala technology). The key feature of this air purifier is the unique innovative multiple filtering system. The treatment of total purification of indoor air showed neither improvements in semen parameters nor variation in reproductive hormones (P = N.S.), but induced an evident increase (P < 0.03 and more) in seminal leucocytic concentrations. Within the limits due to the small sample of subjects recruited, the sole purification of indoor air does not seem enough to improve semen quality, although the increase in leucocytic concentrations could indicate an activation of the role of immunosurveillance in a purified indoor air environment.

  8. Removal of emerging micropollutants from water using cyclodextrin.

    PubMed

    Nagy, Zsuzsanna Magdolna; Molnár, Mónika; Fekete-Kertész, Ildikó; Molnár-Perl, Ibolya; Fenyvesi, Éva; Gruiz, Katalin

    2014-07-01

    Small scale laboratory experiment series were performed to study the suitability of a cyclodextrin-based sorbent (ß-cyclodextrin bead polymer, BCDP) for modelling the removal of micropollutants from drinking water and purified waste water using simulated inflow test solutions containing target analytes (ibuprofen, naproxen, ketoprofen, bisphenol-A, diclofenac, β-estradiol, ethinylestradiol, estriol, cholesterol at 2-6 μg/L level). This work was focused on the preliminary evaluation of BCDP as a sorbent in two different model systems (filtration and fluidization) applied for risk reduction of emerging micropollutants. For comparison different filter systems combined with various sorbents (commercial filter and activated carbon) were applied and evaluated in the filtration experiment series. The spiked test solution (inflow) and the treated outflows were characterized by an integrated methodology including chemical analytical methods gas chromatography-tandem mass spectrometry (GC-MS/MS) and various environmental toxicity tests to determine the efficiency and selectivity of the applied sorbents. Under experimental conditions the cyclodextrin-based filters used for purification of drinking water in most cases were able to absorb more than 90% of the bisphenol-A and of the estrogenic compounds. Both the analytical chemistry and toxicity results showed efficient elimination of these pollutants. Especially the toxicity of the filtrate decreased considerably. Laboratory experiment modelling post-purification of waste water was also performed applying fluidization technology by ß-cyclodextrin bead polymer. The BCDP removed efficiently from the spiked test solution most of the micropollutants, especially the bisphenol-A (94%) and the hormones (87-99%) The results confirmed that the BCDP-containing sorbents provide a good solution to water quality problems and they are able to decrease the load and risk posed by micropollutants to the water systems. Copyright © 2014 Elsevier B.V. All rights reserved.

  9. Clinical application of microencapsulated islets: actual prospectives on progress and challenges.

    PubMed

    Calafiore, Riccardo; Basta, Giuseppe

    2014-04-01

    After 25 years of intense pre-clinical work on microencapsulated intraperitoneal islet grafts into non-immunosuppressed diabetic recipients, the application of this procedure to patients with type 1 diabetes mellitus has been a significant step forward. This result, achieved in a few centers worldwide, underlies the safety of biopolymers used for microencapsulation. Without this advance, no permission for human application of microcapsules would have ever been obtained after years of purification technologies applied to the raw alginates. To improve safety of the encapsulated islet graft system, renewed efforts on the capsules' bioengineering, as well as on insulin-producing cells within the capsular membranes, are in progress. It is hoped that advances in these two critical aspects of the cell encapsulation technology will result in wider human application of this system. Copyright © 2013 Elsevier B.V. All rights reserved.

  10. Safety Assessment of Nanotechnology Products

    EPA Science Inventory

    Nanotechnology has important opportunities to affect technological challenges in such diverse areas as electronics, energy, water purification, food storage, and therapeutics. These emerging technologies hold great promise both for global economic growth and a sustainable environ...

  11. A scalable method for O-antigen purification applied to various Salmonella serovars

    PubMed Central

    Micoli, F.; Rondini, S.; Gavini, M.; Pisoni, I.; Lanzilao, L.; Colucci, A.M.; Giannelli, C.; Pippi, F.; Sollai, L.; Pinto, V.; Berti, F.; MacLennan, C.A.; Martin, L.B.; Saul, A.

    2014-01-01

    The surface lipopolysaccharide of gram-negative bacteria is both a virulence factor and a B cell antigen. Antibodies against O-antigen of lipopolysaccharide may confer protection against infection, and O-antigen conjugates have been designed against multiple pathogens. Here, we describe a simplified methodology for extraction and purification of the O-antigen core portion of Salmonella lipopolysaccharide, suitable for large-scale production. Lipopolysaccharide extraction and delipidation are performed by acetic acid hydrolysis of whole bacterial culture and can take place directly in a bioreactor, without previous isolation and inactivation of bacteria. Further O-antigen core purification consists of rapid filtration and precipitation steps, without using enzymes or hazardous chemicals. The process was successfully applied to various Salmonella enterica serovars (Paratyphi A, Typhimurium, and Enteritidis), obtaining good yields of high-quality material, suitable for conjugate vaccine preparations. PMID:23142430

  12. Recovery and purification process development for monoclonal antibody production

    PubMed Central

    Ma, Junfen; Winter, Charles; Bayer, Robert

    2010-01-01

    Hundreds of therapeutic monoclonal antibodies (mAbs) are currently in development, and many companies have multiple antibodies in their pipelines. Current methodology used in recovery processes for these molecules are reviewed here. Basic unit operations such as harvest, Protein A affinity chromatography and additional polishing steps are surveyed. Alternative processes such as flocculation, precipitation and membrane chromatography are discussed. We also cover platform approaches to purification methods development, use of high throughput screening methods, and offer a view on future developments in purification methodology as applied to mAbs. PMID:20647768

  13. Two-step purification of scutellarin from Erigeron breviscapus (vant.) Hand. Mazz. by high-speed counter-current chromatography.

    PubMed

    Gao, Min; Gu, Ming; Liu, Chun-Zhao

    2006-07-11

    Scutellarin, a flavone glycoside, popularly applied for the treatment of cardiopathy, has been purified in two-step purification by high-speed counter-current chromatography (HSCCC) from Erigeron breviscapus (vant.) Hand. Mazz. (Deng-zhan-hua in Chinese), a well-known traditional Chinese medicinal plant for heart disease. Two solvent systems, n-hexane-ethyl acetate-methanol-acetic acid-water (1:6:1.5:1:4, v/v/v/v/v) and ethyl acetate-n-butanol-acetonitrile-0.1% HCl (5:2:5:10, v/v/v/v) were used for the two-step purification. The purity of the collected fraction of scutellarin was 95.6%. This study supplies a new alternative method for purification of scutellarin.

  14. Aptamer-based downstream processing of his-tagged proteins utilizing magnetic beads.

    PubMed

    Kökpinar, Öznur; Walter, Johanna-Gabriela; Shoham, Yuval; Stahl, Frank; Scheper, Thomas

    2011-10-01

    Aptamers are synthetic nucleic acid-based high affinity ligands that are able to capture their corresponding target via molecular recognition. Here, aptamer-based affinity purification for His-tagged proteins was developed. Two different aptamers directed against the His-tag were immobilized on magnetic beads covalently. The resulting aptamer-modified magnetic beads were characterized and successfully applied for purification of different His-tagged proteins from complex E. coli cell lysates. Purification effects comparable to conventional immobilized metal affinity chromatography were achieved in one single purification step. Moreover, we have investigated the possibility to regenerate and reuse the aptamer-modified magnetic beads and have shown their long-term stability over a period of 6 months. Copyright © 2011 Wiley Periodicals, Inc.

  15. Addition of chlorine during water purification reduces iodine content of drinking water and contributes to iodine deficiency.

    PubMed

    Samson, L; Czegeny, I; Mezosi, E; Erdei, A; Bodor, M; Cseke, B; Burman, K D; Nagy, E V

    2012-01-01

    Drinking water is the major natural source of iodine in many European countries. In the present study, we examined possible sites of iodine loss during the usual water purification process.Water samples from 6 sites during the technological process were taken and analyzed for iodine content. Under laboratory circumstances, prepared iodine in water solution has been used as a model to test the effect of the presence of chlorine. Samples from the purification sites revealed that in the presence of chlorine there is a progressive loss of iodine from the water. In the chlorine concentrations employed in the purification process, 24-h chlorine exposure eliminated more than 50% of iodine when the initial iodine concentration was 250 μg/l or less. Iodine was completely eliminated if the starting concentration was 16 μg/l.We conclude that chlorine used during water purification may be a major contributor to iodine deficiency in European communities.

  16. Approaches to High-Performance Preparative Chromatography of Proteins

    NASA Astrophysics Data System (ADS)

    Sun, Yan; Liu, Fu-Feng; Shi, Qing-Hong

    Preparative liquid chromatography is widely used for the purification of chemical and biological substances. Different from high-performance liquid chromatography for the analysis of many different components at minimized sample loading, high-performance preparative chromatography is of much larger scale and should be of high resolution and high capacity at high operation speed and low to moderate pressure drop. There are various approaches to this end. For biochemical engineers, the traditional way is to model and optimize a purification process to make it exert its maximum capability. For high-performance separations, however, we need to improve chromatographic technology itself. We herein discuss four approaches in this review, mainly based on the recent studies in our group. The first is the development of high-performance matrices, because packing material is the central component of chromatography. Progress in the fabrication of superporous materials in both beaded and monolithic forms are reviewed. The second topic is the discovery and design of affinity ligands for proteins. In most chromatographic methods, proteins are separated based on their interactions with the ligands attached to the surface of porous media. A target-specific ligand can offer selective purification of desired proteins. Third, electrochromatography is discussed. An electric field applied to a chromatographic column can induce additional separation mechanisms besides chromatography, and result in electrokinetic transport of protein molecules and/or the fluid inside pores, thus leading to high-performance separations. Finally, expanded-bed adsorption is described for process integration to reduce separation steps and process time.

  17. EVALUATION OF USFILTER CORPORATION'S RETEC® MODEL SCP-6 SEPARATED CELL PURIFICATION SYSTEM FOR CHROMIC ACID ANODIZE BATH SOLUTION

    EPA Science Inventory

    The USEPA has created the Environmental Technology Verification (ETV) Program to facilitate the deployment of innovative or improved environmental technologies through performance verification and dissemination of information. The ETV P2 Metal Finishing Technologies (ETV-MF) Prog...

  18. Magneto-capillary valve for integrated purification and enrichment of nucleic acids and proteins.

    PubMed

    den Dulk, Remco C; Schmidt, Kristiane A; Sabatté, Gwénola; Liébana, Susana; Prins, Menno W J

    2013-01-07

    We describe the magneto-capillary valve (MCV) technology, a flexible approach for integrated biological sample preparation within the concept of stationary microfluidics. Rather than moving liquids in a microfluidic device, discrete units of liquid are present at fixed positions in the device and magnetic particles are actuated between the fluids. The MCV concept is characterized by the use of two planar surfaces at a capillary mutual distance, with specific features to confine the fluids by capillary forces, and the use of a gas or a phase-change material separating the stationary aqueous liquids. We have studied the physics of magneto-capillary valving by quantifying the magnetic force as a function of time and position, which reveals the balance of magnetic, capillary and frictional forces in the system. By purification experiments with a fluorescent tracer we have measured the amount of co-transported liquid, which is a key parameter for efficient purification. To demonstrate the versatility of the technology, several MCV device architectures were tested in a series of biological assays, showing the purification and enrichment of nucleic acids and proteins. Target recovery comparable to non-miniaturized commercial kits was observed for the extraction of DNA from human cells in buffer, using a device architecture with patterned air valves. Experiments using an enrichment module and patterned air valves demonstrate a 40-fold effective enrichment of DNA in buffer. DNA was also successfully purified from blood plasma using paraffin phase-change valves. Finally, the enrichment of a protein biomarker (prostate-specific antigen) using geometrical air valves resulted in a 7-fold increase of detection signal. The MCV technology is versatile, offers extensive freedom for the design of fully integrated systems, and is expected to be manufacturable in a cost-effective way. We conclude that the MCV technology can become an important enabling technology for point-of-care systems with sample in-result out performance.

  19. Arming Technology in Yeast-Novel Strategy for Whole-cell Biocatalyst and Protein Engineering.

    PubMed

    Kuroda, Kouichi; Ueda, Mitsuyoshi

    2013-09-09

    Cell surface display of proteins/peptides, in contrast to the conventional intracellular expression, has many attractive features. This arming technology is especially effective when yeasts are used as a host, because eukaryotic modifications that are often required for functional use can be added to the surface-displayed proteins/peptides. A part of various cell wall or plasma membrane proteins can be genetically fused to the proteins/peptides of interest to be displayed. This technology, leading to the generation of so-called "arming technology", can be employed for basic and applied research purposes. In this article, we describe various strategies for the construction of arming yeasts, and outline the diverse applications of this technology to industrial processes such as biofuel and chemical productions, pollutant removal, and health-related processes, including oral vaccines. In addition, arming technology is suitable for protein engineering and directed evolution through high-throughput screening that is made possible by the feature that proteins/peptides displayed on cell surface can be directly analyzed using intact cells without concentration and purification. Actually, novel proteins/peptides with improved or developed functions have been created, and development of diagnostic/therapeutic antibodies are likely to benefit from this powerful approach.

  20. Future of antibody purification.

    PubMed

    Low, Duncan; O'Leary, Rhona; Pujar, Narahari S

    2007-03-15

    Antibody purification seems to be safely ensconced in a platform, now well-established by way of multiple commercialized antibody processes. However, natural evolution compels us to peer into the future. This is driven not only by a large, projected increase in the number of antibody therapies, but also by dramatic improvements in upstream productivity, and process economics. Although disruptive technologies have yet escaped downstream processes, evolution of the so-called platform is already evident in antibody processes in late-stage development. Here we perform a wide survey of technologies that are competing to be part of that platform, and provide our [inherently dangerous] assessment of those that have the most promise.

  1. Forward Osmosis in Wastewater Treatment Processes.

    PubMed

    Korenak, Jasmina; Basu, Subhankar; Balakrishnan, Malini; Hélix-Nielsen, Claus; Petrinic, Irena

    2017-01-01

    In recent years, membrane technology has been widely used in wastewater treatment and water purification. Membrane technology is simple to operate and produces very high quality water for human consumption and industrial purposes. One of the promising technologies for water and wastewater treatment is the application of forward osmosis. Essentially, forward osmosis is a process in which water is driven through a semipermeable membrane from a feed solution to a draw solution due to the osmotic pressure gradient across the membrane. The immediate advantage over existing pressure driven membrane technologies is that the forward osmosis process per se eliminates the need for operation with high hydraulic pressure and forward osmosis has low fouling tendency. Hence, it provides an opportunity for saving energy and membrane replacement cost. However, there are many limitations that still need to be addressed. Here we briefly review some of the applications within water purification and new developments in forward osmosis membrane fabrication.

  2. Rapid Two-Step Procedure for Large-Scale Purification of Pediocin-Like Bacteriocins and Other Cationic Antimicrobial Peptides from Complex Culture Medium

    PubMed Central

    Uteng, Marianne; Hauge, Håvard Hildeng; Brondz, Ilia; Nissen-Meyer, Jon; Fimland, Gunnar

    2002-01-01

    A rapid and simple two-step procedure suitable for both small- and large-scale purification of pediocin-like bacteriocins and other cationic peptides has been developed. In the first step, the bacterial culture was applied directly on a cation-exchange column (1-ml cation exchanger per 100-ml cell culture). Bacteria and anionic compounds passed through the column, and cationic bacteriocins were subsequently eluted with 1 M NaCl. In the second step, the bacteriocin fraction was applied on a low-pressure, reverse-phase column and the bacteriocins were detected as major optical density peaks upon elution with propanol. More than 80% of the activity that was initially in the culture supernatant was recovered in both purification steps, and the final bacteriocin preparation was more than 90% pure as judged by analytical reverse-phase chromatography and capillary electrophoresis. PMID:11823243

  3. Using a Water Purification Activity to Teach the Philosophy and Nature of Technology

    ERIC Educational Resources Information Center

    Kruse, Jerrid; Wilcox, Jesse

    2017-01-01

    Next Generation Science Standards (NGSS), with new emphasis on engineering, reflects broadening definitions of scientific and technological literacy. However, engaging in science and engineering practices is necessary, but insufficient, for developing technological literacy. Just as robust scientific literacy includes a deep understanding of the…

  4. [Purifying process of gynostemma pentaphyllum saponins based on "adjoint marker" online control technology and identification of their compositions by UPLC-QTOF-MS].

    PubMed

    Fan, Dong-Dong; Kuang, Yan-Hui; Dong, Li-Hua; Ye, Xiao; Chen, Liang-Mian; Zhang, Dong; Ma, Zhen-Shan; Wang, Jin-Yu; Zhu, Jing-Jing; Wang, Zhi-Min; Wang, De-Qin; Li, Chu-Yuan

    2017-04-01

    To optimize the purification process of gynostemma pentaphyllum saponins (GPS) based on "adjoint marker" online control technology with GPS as the testing index. UPLC-QTOF-MS technology was used for qualitative analysis. "Adjoint marker" online control results showed that the end point of load sample was that the UV absorbance of effluent liquid was equal to half of that of load sample solution, and the absorbance was basically stable when the end point was stable. In UPLC-QTOF-MS qualitative analysis, 16 saponins were identified from GPS, including 13 known gynostemma saponins and 3 new saponins. This optimized method was proved to be simple, scientific, reasonable, easy for online determination, real-time record, and can be better applied to the mass production and automation of production. The results of qualitative analysis indicated that the "adjoint marker" online control technology can well retain main efficacy components of medicinal materials, and provide analysis tools for the process control and quality traceability. Copyright© by the Chinese Pharmaceutical Association.

  5. Water Purification Systems

    NASA Technical Reports Server (NTRS)

    1994-01-01

    Clearwater Pool Technologies employs NASA-developed silver/copper ionization to purify turtle and dolphin tanks, cooling towers, spas, water recycling systems, etc. The pool purifier consists of a microcomputer to monitor water conditions, a pair of metallic electrodes, and a rheostat controller. Ions are generated by passing a low voltage current through the electrodes; the silver ions kill the bacteria, and the copper ions kill algae. This technology has found broad application because it offers an alternative to chemical disinfectants. It was originally developed to purify water on Apollo spacecraft. Caribbean Clear has been using NASA's silver ionization technology for water purification for more than a decade. Two new products incorporate advancements of the basic technology. One is the AquaKing, a system designed for areas with no source of acceptable drinking water. Another is the Caribbean Clear Controller, designed for commercial pool and water park applications where sanitizing is combined with feedback control of pH and an oxidizer, chlorine or bromine. The technology was originally developed to purify water on Apollo spacecraft.

  6. Renewable Energy Production from DoD Installation Solid Wastes by Anaerobic Digestion

    DTIC Science & Technology

    2016-06-01

    and purification of methane -rich biogas was conducted at the US Air Force Academy. Cost and performance of the technology with respect to renewable...SUBJECT TERMS Food waste, FOG, solid waste, anaerobic digestion, methane , biogas, biomethane, biogas purification, vehicle fuel, renewable energy...The project demonstrated the ability to digest these wastes in a controlled and predictable manner to maximize the generation of biogas, a methane

  7. Pool Purification

    NASA Technical Reports Server (NTRS)

    1988-01-01

    Caribbean Clear, Inc. used NASA's silver ion technology as a basis for its automatic pool purifier. System offers alternative approach to conventional purification chemicals. Caribbean Clear's principal markets are swimming pool owners who want to eliminate chlorine and bromine. Purifiers in Caribbean Clear System are same silver ions used in Apollo System to kill bacteria, plus copper ions to kill algae. They produce spa or pool water that exceeds EPA Standards for drinking water.

  8. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Davis, R.; Tao, L.; Scarlata, C.

    This report describes one potential conversion process to hydrocarbon products by way of catalytic conversion of lignocellulosic-derived hydrolysate. This model leverages expertise established over time in biomass deconstruction and process integration research at NREL, while adding in new technology areas for sugar purification and catalysis. The overarching process design converts biomass to die die diesel- and naphtha-range fuels using dilute-acid pretreatment, enzymatic saccharification, purifications, and catalytic conversion focused on deoxygenating and oligomerizing biomass hydrolysates.

  9. A Novel Active-Learning Protein Purification Exercise for Large-Enrollment Introductory Biochemistry Courses Using the CHROM Web Applet

    ERIC Educational Resources Information Center

    Barrette-Ng, Isabelle H.; Usher, Ken C.

    2013-01-01

    The CHROM Web applet has been used to create a new active-learning exercise in which students design a purification scheme for a recombinant protein using ion-exchange chromatography (IEC). To successfully complete the exercise, students are challenged to apply elementary concepts from acid-base chemistry as well as protein and amino acid…

  10. Antigen identification starting from the genome: a "Reverse Vaccinology" approach applied to MenB.

    PubMed

    Palumbo, Emmanuelle; Fiaschi, Luigi; Brunelli, Brunella; Marchi, Sara; Savino, Silvana; Pizza, Mariagrazia

    2012-01-01

    Most of the vaccines available today, albeit very effective, have been developed using traditional "old-style" methodologies. Technologies developed in recent years have opened up new perspectives in the field of vaccinology and novel strategies are now being used to design improved or new vaccines against infections for which preventive measures do not exist. The Reverse Vaccinology (RV) approach is one of the most powerful examples of biotechnology applied to the field of vaccinology for identifying new protein-based vaccines. RV combines the availability of genomic data, the analyzing capabilities of new bioinformatic tools, and the application of high throughput expression and purification systems combined with serological screening assays for a coordinated screening process of the entire genomic repertoire of bacterial, viral, or parasitic pathogens. The application of RV to Neisseria meningitidis serogroup B represents the first success of this novel approach. In this chapter, we describe how this revolutionary approach can be easily applied to any pathogen.

  11. Adsorption and Processes in Spacecraft Environmental Control and Life Support Systems

    NASA Technical Reports Server (NTRS)

    Dall-Bauman, Liese; Finn, John E.; Kliss, Mark (Technical Monitor)

    1997-01-01

    The environmental control and life support system on a spacecraft must maintain a safe and comfortable environment in which the crew can live and work. The system's functions include supplying the crew with oxygen and water, as well as removing carbon dioxide, water vapor, and trace contaminants from cabin air. Although open-loop systems have been used in the past, logistics and safety factors of current and future missions in space make near-complete recycling of the cabin's air and water desirable. The recycling process may include separation and reduction of carbon dioxide, removal of trace gas-phase contaminants, recovery and purification of humidity condensate, purification and polishing of wastewater streams, and other processes. Several of these operations can be performed totally or in part by adsorption processes. Adsorption processes are frequently good candidates for separation and purification in space by virtue of such characteristics as gravity independence, high reliability, relatively high energy efficiency, design flexibility, technological maturity, and regenerability. For these reasons, adsorption has historically played a key role in life support on U.S. and Russian piloted spacecraft. This article focuses on three current spacecraft life support applications that often use adsorption technology: carbon dioxide separation from cabin air, gas-phase trace contaminant control, and potable water recovery from waste streams. In each application, adsorption technology has been selected for use on the International Space Station. The requirements, science, and hardware for each application are discussed. Eventually, human space exploration may lead to construction of planetary habitats. These habitats may have additional applications, such as control of greenhouse gas composition and purification of hydroponic solutions, and may have different requirements and resources available to them, such as gases present in the planetary atmosphere. Adsorption separation and purification processes may continue to fulfill environmental control and life support needs well into the future.

  12. Purification of functionalized DNA origami nanostructures.

    PubMed

    Shaw, Alan; Benson, Erik; Högberg, Björn

    2015-05-26

    The high programmability of DNA origami has provided tools for precise manipulation of matter at the nanoscale. This manipulation of matter opens up the possibility to arrange functional elements for a diverse range of applications that utilize the nanometer precision provided by these structures. However, the realization of functionalized DNA origami still suffers from imperfect production methods, in particular in the purification step, where excess material is separated from the desired functionalized DNA origami. In this article we demonstrate and optimize two purification methods that have not previously been applied to DNA origami. In addition, we provide a systematic study comparing the purification efficacy of these and five other commonly used purification methods. Three types of functionalized DNA origami were used as model systems in this study. DNA origami was patterned with either small molecules, antibodies, or larger proteins. With the results of our work we aim to provide a guideline in quality fabrication of various types of functionalized DNA origami and to provide a route for scalable production of these promising tools.

  13. Carbon nanotube membranes with ultrahigh specific adsorption capacity for water desalination and purification.

    PubMed

    Yang, Hui Ying; Han, Zhao Jun; Yu, Siu Fung; Pey, Kin Leong; Ostrikov, Kostya; Karnik, Rohit

    2013-01-01

    Development of technologies for water desalination and purification is critical to meet the global challenges of insufficient water supply and inadequate sanitation, especially for point-of-use applications. Conventional desalination methods are energy and operationally intensive, whereas adsorption-based techniques are simple and easy to use for point-of-use water purification, yet their capacity to remove salts is limited. Here we report that plasma-modified ultralong carbon nanotubes exhibit ultrahigh specific adsorption capacity for salt (exceeding 400% by weight) that is two orders of magnitude higher than that found in the current state-of-the-art activated carbon-based water treatment systems. We exploit this adsorption capacity in ultralong carbon nanotube-based membranes that can remove salt, as well as organic and metal contaminants. These ultralong carbon nanotube-based membranes may lead to next-generation rechargeable, point-of-use potable water purification appliances with superior desalination, disinfection and filtration properties.

  14. Scalable Electrophysiological Investigation of iPS Cell-Derived Cardiomyocytes Obtained by a Lentiviral Purification Strategy

    PubMed Central

    Friedrichs, Stephanie; Malan, Daniela; Voss, Yvonne; Sasse, Philipp

    2015-01-01

    Disease-specific induced pluripotent stem (iPS) cells can be generated from patients and differentiated into functional cardiomyocytes for characterization of the disease and for drug screening. In order to obtain pure cardiomyocytes for automated electrophysiological investigation, we here report a novel non-clonal purification strategy by using lentiviral gene transfer of a puromycin resistance gene under the control of a cardiac-specific promoter. We have applied this method to our previous reported wild-type and long QT syndrome 3 (LQTS 3)-specific mouse iPS cells and obtained a pure cardiomyocyte population. These cells were investigated by action potential analysis with manual and automatic planar patch clamp technologies, as well as by recording extracellular field potentials using a microelectrode array system. Action potentials and field potentials showed the characteristic prolongation at low heart rates in LQTS 3-specific, but not in wild-type iPS cell-derived cardiomyocytes. Hence, LQTS 3-specific cardiomyocytes can be purified from iPS cells with a lentiviral strategy, maintain the hallmarks of the LQTS 3 disease and can be used for automated electrophysiological characterization and drug screening. PMID:26237021

  15. Extraction and purification methods in downstream processing of plant-based recombinant proteins.

    PubMed

    Łojewska, Ewelina; Kowalczyk, Tomasz; Olejniczak, Szymon; Sakowicz, Tomasz

    2016-04-01

    During the last two decades, the production of recombinant proteins in plant systems has been receiving increased attention. Currently, proteins are considered as the most important biopharmaceuticals. However, high costs and problems with scaling up the purification and isolation processes make the production of plant-based recombinant proteins a challenging task. This paper presents a summary of the information regarding the downstream processing in plant systems and provides a comprehensible overview of its key steps, such as extraction and purification. To highlight the recent progress, mainly new developments in the downstream technology have been chosen. Furthermore, besides most popular techniques, alternative methods have been described. Copyright © 2015 Elsevier Inc. All rights reserved.

  16. 2D nanostructures for water purification: graphene and beyond.

    PubMed

    Dervin, Saoirse; Dionysiou, Dionysios D; Pillai, Suresh C

    2016-08-18

    Owing to their atomically thin structure, large surface area and mechanical strength, 2D nanoporous materials are considered to be suitable alternatives for existing desalination and water purification membrane materials. Recent progress in the development of nanoporous graphene based materials has generated enormous potential for water purification technologies. Progress in the development of nanoporous graphene and graphene oxide (GO) membranes, the mechanism of graphene molecular sieve action, structural design, hydrophilic nature, mechanical strength and antifouling properties and the principal challenges associated with nanopore generation are discussed in detail. Subsequently, the recent applications and performance of newly developed 2D materials such as 2D boron nitride (BN) nanosheets, graphyne, molybdenum disulfide (MoS2), tungsten chalcogenides (WS2) and titanium carbide (Ti3C2Tx) are highlighted. In addition, the challenges affecting 2D nanostructures for water purification are highlighted and their applications in the water purification industry are discussed. Though only a few 2D materials have been explored so far for water treatment applications, this emerging field of research is set to attract a great deal of attention in the near future.

  17. Very large scale monoclonal antibody purification: the case for conventional unit operations.

    PubMed

    Kelley, Brian

    2007-01-01

    Technology development initiatives targeted for monoclonal antibody purification may be motivated by manufacturing limitations and are often aimed at solving current and future process bottlenecks. A subject under debate in many biotechnology companies is whether conventional unit operations such as chromatography will eventually become limiting for the production of recombinant protein therapeutics. An evaluation of the potential limitations of process chromatography and filtration using today's commercially available resins and membranes was conducted for a conceptual process scaled to produce 10 tons of monoclonal antibody per year from a single manufacturing plant, a scale representing one of the world's largest single-plant capacities for cGMP protein production. The process employs a simple, efficient purification train using only two chromatographic and two ultrafiltration steps, modeled after a platform antibody purification train that has generated 10 kg batches in clinical production. Based on analyses of cost of goods and the production capacity of this very large scale purification process, it is unlikely that non-conventional downstream unit operations would be needed to replace conventional chromatographic and filtration separation steps, at least for recombinant antibodies.

  18. High-yield fermentation and a novel heat-precipitation purification method for hydrophobin HGFI from Grifola frondosa in Pichia pastoris.

    PubMed

    Song, Dongmin; Gao, Zhendong; Zhao, Liqiang; Wang, Xiangxiang; Xu, Haijin; Bai, Yanling; Zhang, Xiuming; Linder, Markus B; Feng, Hui; Qiao, Mingqiang

    2016-12-01

    Hydrophobins are proteins produced by filamentous fungi with high natural-surfactant activities and that can self-assemble in interfaces of air-water or solid-water to form amphiphilic membranes. Here, we reported a high-yield fermentation method for hydrophobin HGFI from Grifola frondosa in Pichia pastoris, attaining production of 300 mg/L by keeping the dissolved oxygen level at 15%-25% by turning the methanol-feeding speed. We also developed a novel HGFI-purification method enabling large-scare purification of HGFI, with >90% recovery. Additionally, we observed that hydrophobin HGFI in fermentation broth precipitated at pH < 7.0 and temperatures >90 °C. We also identified the structure and properties of proteins purified by this method through atomic force microscopy, circular dichroism, X-ray photoelectron spectroscopy, and water-contact angle measurement, which is similar to protein purification by ultrafiltration without heating treatment that enables our method to maintain native HGFI structure and properties. Furthermore, the purification method presented here can be applied to large-scale purification of other type I hydrophobins. Copyright © 2016. Published by Elsevier Inc.

  19. Strong ion exchange in centrifugal partition extraction (SIX-CPE): effect of partition cell design and dimensions on purification process efficiency.

    PubMed

    Hamzaoui, Mahmoud; Hubert, Jane; Reynaud, Romain; Marchal, Luc; Foucault, Alain; Renault, Jean-Hugues

    2012-07-20

    The aim of this article was to evaluate the influence of the column design of a hydrostatic support-free liquid-liquid chromatography device on the process efficiency when the strong ion-exchange (SIX) development mode is used. The purification of p-hydroxybenzylglucosinolate (sinalbin) from a crude aqueous extract of white mustard seeds (Sinapis alba L.) was achieved on two types of devices: a centrifugal partition chromatograph (CPC) and a centrifugal partition extractor (CPE). They differ in the number, volume and geometry of their partition cells. The SIX-CPE process was evaluated in terms of productivity and sinalbin purification capability as compared to previously optimized SIX-CPC protocols that were carried out on columns of 200 mL and 5700 mL inner volume, respectively. The objective was to determine whether the decrease in partition cell number, the increase in their volume and the use of a "twin cell" design would induce a significant increase in productivity by applying higher mobile phase flow rate while maintaining a constant separation quality. 4.6g of sinalbin (92% recovery) were isolated from 25 g of a crude white mustard seed extract, in only 32 min and with a purity of 94.7%, thus corresponding to a productivity of 28 g per hour and per liter of column volume (g/h/LV(c)). Therefore, the SIX-CPE process demonstrates promising industrial technology transfer perspectives for the large-scale isolation of ionized natural products. Copyright © 2012 Elsevier B.V. All rights reserved.

  20. Highly efficient purification of protein complexes from mammalian cells using a novel streptavidin-binding peptide and hexahistidine tandem tag system: Application to Bruton's tyrosine kinase

    PubMed Central

    Li, Yifeng; Franklin, Sarah; Zhang, Michael J; Vondriska, Thomas M

    2011-01-01

    Tandem affinity purification (TAP) is a generic approach for the purification of protein complexes. The key advantage of TAP is the engineering of dual affinity tags that, when attached to the protein of interest, allow purification of the target protein along with its binding partners through two consecutive purification steps. The tandem tag used in the original method consists of two IgG-binding units of protein A from Staphylococcus aureus (ProtA) and the calmodulin-binding peptide (CBP), and it allows for recovery of 20–30% of the bait protein in yeast. When applied to higher eukaryotes, however, this classical TAP tag suffers from low yields. To improve protein recovery in systems other than yeast, we describe herein the development of a three-tag system comprised of CBP, streptavidin-binding peptide (SBP) and hexa-histidine. We illustrate the application of this approach for the purification of human Bruton's tyrosine kinase (Btk), which results in highly efficient binding and elution of bait protein in both purification steps (>50% recovery). Combined with mass spectrometry for protein identification, this TAP strategy facilitated the first nonbiased analysis of Btk interacting proteins. The high efficiency of the SBP-His6 purification allows for efficient recovery of protein complexes formed with a target protein of interest from a small amount of starting material, enhancing the ability to detect low abundance and transient interactions in eukaryotic cell systems. PMID:21080425

  1. AN ENVIRONMENTAL AND ECONOMIC COMPARISON OF ION EXCHANGE AND RECENTLY COMMERCIALIZED ELECTROCHEMICAL TECHNOLOGIES FOR THE RECOVERY OF RINSE WATER IN BRIGHT NICKEL PLATING FACILITY

    EPA Science Inventory

    Researchers at USEPA are testing and evaluating two commercial electrochemical technologies for the purification of rinse water and the recovery of copper and nickel from a variety of electroplating processes. One of the investigated technologies is based on the application of hi...

  2. Polymer Based Highly Parallel Nanoscopic Sensors for Rapid Detection of Chemical and Biological Threats

    DTIC Science & Technology

    2007-09-18

    Xuliang Han, PI of Brewer Science, Inc. Subcontract Center for Applied Science & Engineering Missouri State University 901 South National Avenue...Science an effective post-growth purification procedure was developed to reduce the amount of impurities, and several characterization techniques were...CNTs) contain a wide range of impurities from the growth process. At Brewer Science an effective post-growth purification procedure was developed to

  3. Biocapture of CO2 by Different Microalgal-Based Technologies for Biogas Upgrading and Simultaneous Biogas Slurry Purification under Various Light Intensities and Photoperiods

    PubMed Central

    Guo, Pengfei; Zhang, Yuejin; Zhao, Yongjun

    2018-01-01

    Co-cultivation of microalgae and microbes for pollutant removal from sewage is considered as an effective wastewater treatment method. The aim of this study is to screen the optimal photoperiod, light intensity and microalgae co-cultivation method for simultaneously removing nutrients in biogas slurry and capturing CO2 in biogas. The microalgae–fungi pellets are deemed to be a viable option because of their high specific growth rate and nutrient and CO2 removal efficiency under the photoperiod of 14 h light:10 h dark. The order of both the biogas slurry purification and biogas upgrading is ranked the same, that is Chlorella vulgaris–Ganoderma lucidum > Chlorella vulgaris–activated sludge > Chlorella vulgaris under different light intensities. For all cultivation methods, the moderate light intensity of 450 μmol m−2 s−1 is regarded as the best choice. This research revealed that the control of photoperiod and light intensity can promote the biological treatment process of biogas slurry purification and biogas upgrading using microalgal-based technology. PMID:29543784

  4. Biocapture of CO₂ by Different Microalgal-Based Technologies for Biogas Upgrading and Simultaneous Biogas Slurry Purification under Various Light Intensities and Photoperiods.

    PubMed

    Guo, Pengfei; Zhang, Yuejin; Zhao, Yongjun

    2018-03-15

    Abstract : Co-cultivation of microalgae and microbes for pollutant removal from sewage is considered as an effective wastewater treatment method. The aim of this study is to screen the optimal photoperiod, light intensity and microalgae co-cultivation method for simultaneously removing nutrients in biogas slurry and capturing CO₂ in biogas. The microalgae-fungi pellets are deemed to be a viable option because of their high specific growth rate and nutrient and CO 2 removal efficiency under the photoperiod of 14 h light:10 h dark. The order of both the biogas slurry purification and biogas upgrading is ranked the same, that is Chlorella vulga ris - Ganoderma lucidum > Chlorella vulga ris -activated sludge > Chlorella vulgaris under different light intensities. For all cultivation methods, the moderate light intensity of 450 μmol m -2 s -1 is regarded as the best choice. This research revealed that the control of photoperiod and light intensity can promote the biological treatment process of biogas slurry purification and biogas upgrading using microalgal-based technology.

  5. Design and analysis of siloxanes removal by adsorption from landfill gas for waste-to-energy processes.

    PubMed

    Elwell, Anthony C; Elsayed, Nada H; Kuhn, John N; Joseph, Babu

    2018-03-01

    Separation of volatile methyl siloxanes from landfill gas using fixed adsorption beds was modeled with the objective of identifying appropriate technology and the economics associated with this purification step. A general adsorption model assuming plug flow and radial symmetry was developed and used to conduct a parametric sweep of 162 unique cases. The varied parameters were adsorbent type (activated carbon and silica gel), bed height (3.05-9.15 m/10-30 ft), inlet siloxane concentration (5-15 mg/m 3 ), moisture content (0-100% relative humidity at STP or RH), and siloxane tolerance limit (0.094-9.4 mg/m 3 ) that correlated to three distinct energy conversion technologies (electricity production using engines or fuels cells or catalytic conversion to liquid hydrocarbon fuels). Due to the detrimental effect of RH on siloxane absorption, the maximum allowable moisture content of LFG before purification is 50% RH and moisture removal processes are also required. The design calculations using a selected case study show that the adsorption bed height required needed for 6 months minimum breakthrough time for catalytic fuel production is twice that for engine applications. Fuel cell applications require 3 times the bed height compared to engine applications. However, the purification costs amounted to 94%, 16% and 52% of recovered product value for engine, liquefaction, and fuel cell applications, respectively indicating the need for a high value product to justify purification costs. The approaches and conclusions can be extended to specific process conditions for landfill gas purification and to other processes that use biogas produced from waste as a feedstock. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. [Study on extraction and purification process of total ginsenosides from Radix Ginseng].

    PubMed

    Xie, Li-Ling; Ren, Li; Lai, Xian-Sheng; Cao, Jun-Hui; Mo, Quan-Yi; Chen, Wei-Wen

    2009-10-01

    To optimize the technological parameters of the extraction and purification process of total ginsenosides from Radix Ginseng. With the contents of ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1, the orthogonal design was adopted to optimize the extraction process. The purification process was studied by optimizing the elutive ratio of total ginsenosides as the marker. HPLC and spectrophotometer were employed for the study. The optimum conditions were as follows:Using 8 times volume of 75% ethanol extracting for 120 minutes and 2 times, the extraction temperature was 85 degrees C. AB-8 macroporous resin was selected, and the eluant was 4 BV 70% ethanol. The optimal conditions of extracting and purifying the total ginsenosides from Radix Ginseng is feasible.

  7. Entanglement concentration and purification of two-mode squeezed microwave photons in circuit QED

    NASA Astrophysics Data System (ADS)

    Zhang, Hao; Alsaedi, Ahmed; Hayat, Tasawar; Deng, Fu-Guo

    2018-04-01

    We present a theoretical proposal for a physical implementation of entanglement concentration and purification protocols for two-mode squeezed microwave photons in circuit quantum electrodynamics (QED). First, we give the description of the cross-Kerr effect induced between two resonators in circuit QED. Then we use the cross-Kerr media to design the effective quantum nondemolition (QND) measurement on microwave-photon number. By using the QND measurement, the parties in quantum communication can accomplish the entanglement concentration and purification of nonlocal two-mode squeezed microwave photons. We discuss the feasibility of our schemes by giving the detailed parameters which can be realized with current experimental technology. Our work can improve some practical applications in continuous-variable microwave-based quantum information processing.

  8. Rapid purification of diastereoisomers from Piper kadsura using supercritical fluid chromatography with chiral stationary phases.

    PubMed

    Xin, Huaxia; Dai, Zhuoshun; Cai, Jianfeng; Ke, Yanxiong; Shi, Hui; Fu, Qing; Jin, Yu; Liang, Xinmiao

    2017-08-04

    Supercritical fluid chromatography (SFC) with chiral stationary phases (CSPs) is an advanced solution for the separation of achiral compounds in Piper kadsura. Analogues and stereoisomers are abundant in natural products, but there are obstacles in separation using conventional method. In this paper, four lignan diastereoisomers, (-)-Galbelgin, (-)-Ganschisandrin, Galgravin and (-)-Veraguensin, from Piper kadsura were separated and purified by chiral SFC. Purification strategy was designed, considering of the compound enrichment, sample purity and purification throughput. Two-step achiral purification method on chiral preparative columns with stacked automated injections was developed. Unconventional mobile phase modifier dichloromethane (DCM) was applied to improve the sample solubility. Four diastereoisomers was prepared at the respective weight of 103.1mg, 10.0mg, 152.3mg and 178.6mg from 710mg extract with the purity of greater than 98%. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Antimicrobial Peptide Production and Purification.

    PubMed

    Suda, Srinivas; Field, Des; Barron, Niall

    2017-01-01

    Antimicrobial peptides (AMPs) are natural defense compounds which are synthesized as ribosomal gene-encoded pre-peptides and produced by all living organisms. AMPs are small peptides, usually cationic and typically have hydrophobic residues which interact with cell membranes and have either a narrow or broad spectrum of biological activity. AMPs are isolated from the natural host or heterologously expressed in other hosts such as Escherichia coli. The proto-typical lantibiotic Nisin is a widely used AMP that is produced by the food-grade organism Lactococcus lactis. Although AMP production and purification procedures require optimization for individual AMPs, the Nisin production and purification protocol outlined in this chapter can be easily applied with minor modifications for the production and purification of other lantibiotics or AMPs. While Nisin is produced and secreted into the supernatant, steps to recover Nisin from both cell-free supernatant and cell pellet are outlined in detail.

  10. Rapid and Scalable Characterization of CRISPR Technologies Using an E. coli Cell-Free Transcription-Translation System.

    PubMed

    Marshall, Ryan; Maxwell, Colin S; Collins, Scott P; Jacobsen, Thomas; Luo, Michelle L; Begemann, Matthew B; Gray, Benjamin N; January, Emma; Singer, Anna; He, Yonghua; Beisel, Chase L; Noireaux, Vincent

    2018-01-04

    CRISPR-Cas systems offer versatile technologies for genome engineering, yet their implementation has been outpaced by ongoing discoveries of new Cas nucleases and anti-CRISPR proteins. Here, we present the use of E. coli cell-free transcription-translation (TXTL) systems to vastly improve the speed and scalability of CRISPR characterization and validation. TXTL can express active CRISPR machinery from added plasmids and linear DNA, and TXTL can output quantitative dynamics of DNA cleavage and gene repression-all without protein purification or live cells. We used TXTL to measure the dynamics of DNA cleavage and gene repression for single- and multi-effector CRISPR nucleases, predict gene repression strength in E. coli, determine the specificities of 24 diverse anti-CRISPR proteins, and develop a fast and scalable screen for protospacer-adjacent motifs that was successfully applied to five uncharacterized Cpf1 nucleases. These examples underscore how TXTL can facilitate the characterization and application of CRISPR technologies across their many uses. Copyright © 2017 Elsevier Inc. All rights reserved.

  11. Innovative Process for Comprehensive Treatment of Liquid Radioactive Waste - 12551

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Penzin, R.A.; Sarychev, G.A.

    This paper presents the results of research activities aimed at creation of a principally new LRW distilling treatment method. The new process is based on the instantaneous evaporation method widely used in distillation units. The main difference of the proposed process is that the vapor condensation is conducted without using heat exchangers in practically ideal mode by way of direct contacting in a vapor-liquid system. This process is conducted in a specially designed ejector unit in supersonic mode. Further recuperation of excess heat of vaporization is carried out in a standard heat exchanger. Such an arrangement of the process, togethermore » with use of the barometric height principle, allows to carry out LRW evaporation under low temperatures, which enables to use excess heat from NPS for heating initial LRW. Thermal calculations and model experiments have revealed that, in this case, the expenditure of energy for LRW treatment by distilling will not exceed 3 kilowatt-hour/m{sup 3}, which is comparable with the reverse-osmosis desalination method. Besides, the proposed devices are 4 to 5 times less metal-intensive than standard evaporation units. These devices are also characterized by versatility. Experiments have revealed that the new method can be used for evaporation of practically any types of LRW, including those containing a considerable amount of oil products. Owing to arrangement of the evaporation process at low temperatures, the new devices are not sensitive to 'scale formation'. This is why, they can be used for concentrating brines of up to 500-600 g/l. New types of such evaporating devices can be required both for LRW treatment processes at nuclear-power plants under design and for treating 'non-standard' LRW with complex physicochemical and radionuclide composition resulting from the disaster at the Fukushima I Nuclear Power Plant.) As a result of accidents at nuclear energy objects, as it has recently happened at NPP 'Fukushima-1', personnel faces the necessity to take emergency measures and to use marine water for cooling of reactor zone in contravention of the technological regulations. In these cases significant amount of liquid radioactive wastes of complex physicochemical composition is being generated, the purification of which by traditional methods is close to impossible. According to the practice of elimination of the accident after-effects at NPP 'Fukushima' there are still no technical means for the efficient purification of liquid radioactive wastes of complex composition like marine water from radionuclides. Therefore development of state-of-the-art highly efficient facilities capable of fast and safe purification of big amounts of liquid radioactive wastes of complex physicochemical composition from radionuclides turns to be utterly topical problem. Cesium radionuclides, being extremely dangerous for the environment, present over 90% of total radioactivity contained in liquid radioactive wastes left as a result of accidents at nuclear power objects. For the purpose of radiation accidents aftereffects liquidation VNIIHT proposes to create a plant for LRW reprocessing, consisting of 4 major technological modules: Module of LRW pretreatment to remove mechanical and organic impurities including oil products; Module of sorption purification of LWR by means of selective inorganic sorbents; Module of reverse osmotic purification and desalination; Module of deep evaporation of LRW concentrates. The first free modules are based on completed technological and designing concepts implemented by VNIIHT in the framework of LLRW Project in the period of 2000-2001 in Russia for comprehensive treatment of LWR of atomic fleet. These industrial plants proved to be highly efficient and secure during their long operation life. Module of deep evaporation is a new technological development. It will ensure conduction of evaporation and purification of LRW of different physicochemical composition, including those containing hardness salts, resulted in generation of LRW concentrate 300-600 g/l. The method is based on utilization of supersonic ejector for intensification of thermal physic processes and performance of evaporation in brine recycling mode. All proposed technological solutions are totally based on patented Russian developments. Proposed work will allow to construct modular plants, which will be totally prepared for efficient purification of any types of liquid radioactive wastes from radionuclides in case of force majeure. According to proposed scheme concentration level of cesium radionuclides in safe-for-storage form will make up not less than 5000. With respect to purification from cesium radionuclides of liquid radioactive wastes stored at NPP 'Fukushima' about 10 t of inorganic sorbents, loaded in 160 protective filter-containers, will be required for solving this problem. The amount of secondary wastes will be reduced approximately in 5 times in comparison with traditional schemes, applied in purification of secondary LRW of Fukushima-1 by Areva (France) and Kurion (USA) companies. All units of modular plants will be constructed and manufactured as totally automated, providing their twenty-four-hour safe operation. Modular design will ensure efficiency and let optimize the costs of secondary LRW treatment. In order to ensure off-line operation in emergency conditions the plant should be equipped with auxiliary modules: energy and ventilation ones. Under normal conditions these modules can be stored in 'mothballed' condition at special warehouses under the authority of federal bodies. It will be reasonable to choose required transport facilities, the most suitable for transportation of modules to target destination beforehand, using vessel classification list.« less

  12. Standing surface acoustic wave technology applied for micro-particle concentration in oil

    NASA Astrophysics Data System (ADS)

    Wang, Ziping; Xue, Xian; Luo, Ying; Yuan, Fuh-Gwo

    2018-03-01

    Oil lubrication plays an important role in a variety of mechanical equipment. The traditional purification method is difficult to remove the tiny impurity size of 5-15 μm. Three different types of the transducers and its preparation methods were used in the experiment. The phenomenon that the impurity particles in viscous fluid by the acoustic radiation force was moved the wave node position and focused on the center line was observed by the super-depth microscope. The influence factors of the produced SSAW, particle force condition and movement track were analyzed. The experimental results show that the interdigital transducer can be used to generate SSAW, so as to achieve the separation effect of oil and suspended particles.

  13. Water Purifier

    NASA Technical Reports Server (NTRS)

    1992-01-01

    The Floatron water purifier combines two space technologies - ionization for water purification and solar electric power generation. The water purification process involves introducing ionized minerals that kill microorganisms like algae and bacteria. The 12 inch unit floats in a pool while its solar panel collects sunlight that is converted to electricity. The resulting current energizes a specially alloyed mineral electrode below the waterline, causing release of metallic ions into the water. The electrode is the only part that needs replacing, and water purified by the system falls within EPA drinking water standards.

  14. Integrated Method for Purification and Single-Particle Characterization of Lentiviral Vector Systems by Size Exclusion Chromatography and Tunable Resistive Pulse Sensing.

    PubMed

    Heider, Susanne; Muzard, Julien; Zaruba, Marianne; Metzner, Christoph

    2017-07-01

    Elements derived from lentiviral particles such as viral vectors or virus-like particles are commonly used for biotechnological and biomedical applications, for example in mammalian protein expression, gene delivery or therapy, and vaccine development. Preparations of high purity are necessary in most cases, especially for clinical applications. For purification, a wide range of methods are available, from density gradient centrifugation to affinity chromatography. In this study we have employed size exclusion columns specifically designed for the easy purification of extracellular vesicles including exosomes. In addition to viral marker protein and total protein analysis, a well-established single-particle characterization technology, termed tunable resistive pulse sensing, was employed to analyze fractions of highest particle load and purity and characterize the preparations by size and surface charge/electrophoretic mobility. With this study, we propose an integrated platform combining size exclusion chromatography and tunable resistive pulse sensing for monitoring production and purification of viral particles.

  15. Technological assumptions for biogas purification.

    PubMed

    Makareviciene, Violeta; Sendzikiene, Egle

    2015-01-01

    Biogas can be used in the engines of transport vehicles and blended into natural gas networks, but it also requires the removal of carbon dioxide, hydrogen sulphide, and moisture. Biogas purification process flow diagrams have been developed for a process enabling the use of a dolomite suspension, as well as for solutions obtained by the filtration of the suspension, to obtain biogas free of hydrogen sulphide and with a carbon dioxide content that does not exceed 2%. The cost of biogas purification was evaluated on the basis of data on biogas production capacity and biogas production cost obtained from local water treatment facilities. It has been found that, with the use of dolomite suspension, the cost of biogas purification is approximately six times lower than that in the case of using a chemical sorbent such as monoethanolamine. The results showed travelling costs using biogas purified by dolomite suspension are nearly 1.5 time lower than travelling costs using gasoline and slightly lower than travelling costs using mineral diesel fuel.

  16. Optimization of a micro-scale, high throughput process development tool and the demonstration of comparable process performance and product quality with biopharmaceutical manufacturing processes.

    PubMed

    Evans, Steven T; Stewart, Kevin D; Afdahl, Chris; Patel, Rohan; Newell, Kelcy J

    2017-07-14

    In this paper, we discuss the optimization and implementation of a high throughput process development (HTPD) tool that utilizes commercially available micro-liter sized column technology for the purification of multiple clinically significant monoclonal antibodies. Chromatographic profiles generated using this optimized tool are shown to overlay with comparable profiles from the conventional bench-scale and clinical manufacturing scale. Further, all product quality attributes measured are comparable across scales for the mAb purifications. In addition to supporting chromatography process development efforts (e.g., optimization screening), comparable product quality results at all scales makes this tool is an appropriate scale model to enable purification and product quality comparisons of HTPD bioreactors conditions. The ability to perform up to 8 chromatography purifications in parallel with reduced material requirements per run creates opportunities for gathering more process knowledge in less time. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  17. Tetanus toxoid purification: chromatographic procedures as an alternative to ammonium-sulphate precipitation.

    PubMed

    Stojićević, Ivana; Dimitrijević, Ljiljana; Dovezenski, Nebojša; Živković, Irena; Petrušić, Vladimir; Marinković, Emilija; Inić-Kanada, Aleksandra; Stojanović, Marijana

    2011-08-01

    Given an existing demand to establish a process of tetanus vaccine production in a way that allows its complete validation and standardization, this paper focuses on tetanus toxoid purification step. More precisely, we were looking at a possibility to replace the widely used ammonium-sulphate precipitation by a chromatographic method. Based on the tetanus toxin's biochemical characteristics, we have decided to examine the possibility of tetanus toxoid purification by hydrophobic chromatography, and by chromatographic techniques based on interaction with immobilized metal ions, i.e. chelating chromatography and immobilized metal affinity chromatography. We used samples obtained from differently fragmented crude tetanus toxins by formaldehyde treatment (assigned as TTd-A and TTd-B) as starting material for tetanus toxoid purification. Obtained results imply that purification of tetanus toxoid by hydrophobic chromatography represents a good alternative to ammonium-sulphate precipitation. Tetanus toxoid preparations obtained by hydrophobic chromatography were similar to those obtained by ammonium-sulphate precipitation in respect to yield, purity and immunogenicity. In addition, their immunogenicity was similar to standard tetanus toxoid preparation (NIBSC, Potters Bar, UK). Furthermore, the characteristics of crude tetanus toxin preparations had the lowest impact on the final purification product when hydrophobic chromatography was the applied method of tetanus toxoid purification. On the other hand, purifications of tetanus toxoid by chelating chromatography or immobilized metal affinity chromatography generally resulted in a very low yield due to not satisfactory tetanus toxoid binding to the column, and immunogenicity of the obtained tetanus toxoid-containing preparations was poor. Copyright © 2011 Elsevier B.V. All rights reserved.

  18. Purification of recombinant Aβ(1-42) and pGlu-Aβ(3-42) using preparative SDS-PAGE.

    PubMed

    Spahn, Claudia; Wermann, Michael; Eichentopf, Rico; Hause, Gerd; Schlenzig, Dagmar; Schilling, Stephan

    2017-08-01

    Recombinant expression and purification of amyloid peptides represents a common basis for investigating the molecular mechanisms of amyloid formation and toxicity. However, the isolation of the recombinant peptides is hampered by inefficient separation from contaminants such as the fusion protein required for efficient expression in E. coli. Here, we present a new approach for the isolation of highly purified Aβ(1-42) and pGlu-Aβ(3-42), which is based on a separation using preparative SDS-PAGE. The method relies on the purification of the Aβ fusion protein by affinity chromatography followed by preparative SDS-PAGE under reducing conditions and subsequent removal of detergents by precipitation. The application of preparative SDS-PAGE represents the key step to isolate highly pure recombinant Aβ, which has been applied for characterization of aggregation and toxicity. Thereby, the yield of the purification strategy was  >60%. To the best of our knowledge, this is the first description of an electrophoresis-based method for purification of a recombinant Aβ peptide. Therefore, the method might be of interest for isolation of other amyloid peptides, which are critical for conventional purification strategies due to their aggregation propensity. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Soil-based filtration technology for air purification: potentials for environmental and space life support application

    NASA Astrophysics Data System (ADS)

    Nelson, Mark; Bohn, Hinrich

    Soil biofiltration, also known as Soil bed reactor (SBR), technology was originally developed in Germany to take advantage of the diversity in microbial mechanisms to control gases producing malodor in industrial processes. The approach has since gained wider international acceptance and seen numerous improvements, for example, by the use of high-organic compost beds to maximize microbial processes. This paper reviews the basic mechanisms which underlay soil processes involved in air purification, advantages and limitations of the technology and the cur-rent research status of the approach. Soil biofiltration has lower capital and operating/energetic costs than conventional technologies and is well adapted to handle contaminants in moderate concentrations. The systems can be engineered to optimize efficiency though manipulation of temperature, pH, moisture content, soil organic matter and airflow rates. SBR technology was modified for application in the Biosphere 2 project, which demonstrated in preparatory research with a number of closed system testbeds that soil could also support crop plants while also serving as soil filters with air pumps to push air through the soil. This Biosphere 2 research demonstrated in several closed system testbeds that a number of important trace gases could be kept under control and led to the engineering of the entire agricultural soil of Biosphere 2 to serve as a soil filtration unit for the facility. Soil biofiltration, coupled with food crop produc-tion, as a component of bioregenerative space life support systems has the advantages of lower energy use and avoidance of the consumables required for other air purification approaches. Expanding use of soil biofiltration can aid a number of environmental applications, from the mitigation of indoor air pollution, improvement of industrial air emissions and prevention of accidental release of toxic gases.

  20. The Radio Frequency Fragment Separator for Rare Isotope Beams at the NSCL

    NASA Astrophysics Data System (ADS)

    Stoker, Joshua; Andreev, Vladimir; Bazin, Daniel; Becerril, Ana; Doleans, Marc; Gorelov, Dimitry; Glennon, Patrick; Grimm, Terry; Lawton, Don; Mantica, Paul; Marti, Felix; Ottarson, Jack; Schatz, Hendrik; Vincent, John; Wagner, Jim; Wu, Xiaoyu; Zeller, Al

    2006-10-01

    Secondary beams at the National Superconducting Cyclotron Laboratory (NSCL) are separated through a combined application of magnetic rigidity and energy loss filtering. Design and construction of a Radio Frequency Fragment Separator (RFFS) for further beam purification is underway. The RFFS will apply a time-varying electromagnetic field to induce transverse beam separation. This method relies on velocity differences of the beam species to selectivey apply separation to unwanted fragments. The technical design of the RFFS and the expected purification of exotic beams are shown in detail[1]. [1] Gorelev, D. et al., ``RF Kicker System for Secondary Beams at the NSCL'' Proc of Part Accel Conf 2005, Knoxville, TN

  1. Postextraction Separation, On-Board Storage, and Catalytic Conversion of Methane in Natural Gas: A Review.

    PubMed

    Saha, Dipendu; Grappe, Hippolyte A; Chakraborty, Amlan; Orkoulas, Gerassimos

    2016-10-12

    In today's perspective, natural gas has gained considerable attention, due to its low emission, indigenous availability, and improvement in the extraction technology. Upon extraction, it undergoes several purification protocols including dehydration, sweetening, and inert rejection. Although purification is a commercially established technology, several drawbacks of the current process provide an essential impetus for developing newer separation protocols, most importantly, adsorption and membrane separation. This Review summarizes the needs of natural gas separation, gives an overview of the current technology, and provides a detailed discussion of the progress in research on separation and purification of natural gas including the benefits and drawbacks of each of the processes. The transportation sector is another growing sector of natural gas utilization, and it requires an efficient and safe on-board storage system. Compressed natural gas (CNG) and liquefied natural gas (LNG) are the most common forms in which natural gas can be stored. Adsorbed natural gas (ANG) is an alternate storage system of natural gas, which is advantageous as compared to CNG and LNG in terms of safety and also in terms of temperature and pressure requirements. This Review provides a detailed discussion on ANG along with computation predictions. The catalytic conversion of methane to different useful chemicals including syngas, methanol, formaldehyde, dimethyl ether, heavier hydrocarbons, aromatics, and hydrogen is also reviewed. Finally, direct utilization of methane onto fuel cells is also discussed.

  2. Purification of phage display-modified bacteriophage T4 by affinity chromatography

    PubMed Central

    2011-01-01

    Background Affinity chromatography is one of the most efficient protein purification strategies. This technique comprises a one-step procedure with a purification level in the order of several thousand-fold, adaptable for various proteins, differentiated in their size, shape, charge, and other properties. The aim of this work was to verify the possibility of applying affinity chromatography in bacteriophage purification, with the perspective of therapeutic purposes. T4 is a large, icosahedral phage that may serve as an efficient display platform for foreign peptides or proteins. Here we propose a new method of T4 phage purification by affinity chromatography after its modification with affinity tags (GST and Histag) by in vivo phage display. As any permanent introduction of extraneous DNA into a phage genome is strongly unfavourable for medical purposes, integration of foreign motifs with the phage genome was not applied. The phage was propagated in bacteria expressing fusions of the phage protein Hoc with affinity tags from bacterial plasmids, independently from the phage expression system. Results Elution profiles of phages modified with the specific affinity motifs (compared to non-specific phages) document their binding to the affinity resins and effective elution with standard competitive agents. Non-specific binding was also observed, but was 102-105 times weaker than the specific one. GST-modified bacteriophages were also effectively released from glutathione Sepharose by proteolytic cleavage. The possibility of proteolytic release was designed at the stage of expression vector construction. Decrease in LPS content in phage preparations was dependent on the washing intensity; intensive washing resulted in preparations of 11-40 EU/ml. Conclusions Affinity tags can be successfully incorporated into the T4 phage capsid by the in vivo phage display technique and they strongly elevate bacteriophage affinity to a specific resin. Affinity chromatography can be considered as a new phage purification method, appropriate for further investigations and development. PMID:21627821

  3. Affinity precipitation of human serum albumin using a thermo-response polymer with an L-thyroxin ligand.

    PubMed

    Ding, Zhaoyang; Cao, Xuejun

    2013-12-17

    Affinity precipitation has been reported as a potential technology for the purification of proteins at the early stage of downstream processing. The technology could be achieved using reversible soluble-insoluble polymers coupled with an affinity ligand to purify proteins from large volumes of dilute solution material such as fermentation broths or plasma. In this study, a thermo-response polymer was synthesized using N-methylol acrylamide, N-isopropyl acrylamide and butyl acrylate as monomers. The molecular weight of the polymer measured by the viscosity method was 3.06 × 104 Da and the lower critical solution temperature (LCST) was 28.0°C.The recovery of the polymer above the LCST was over 95.0%. Human serum albumin (HSA) is the most abundant protein in the human serum system, and it has important functions in the human body. High purity HSA is required in pharmaceuticals. Safe and efficient purification is a crucial process during HSA production. A thermo-response polymer was synthesized and L-thyroxin immobilized on the polymer as an affinity ligand to enable affinity precipitation of HSA. The LCST of the affinity polymer was 31.0°C and the recovery was 99.6% of its original amount after recycling three times. The optimal adsorption condition was 0.02 M Tris-HCl buffer (pH 7.0) and the HSA adsorption capacity was 14.9 mg/g polymer during affinity precipitation. Circular dichroism spectra and a ForteBio Octet system were used to analyze the interactions between the affinity polymer and HSA during adsorption and desorption. The recovery of total HSA by elution with 1.0 mol/L NaSCN was 93.6%. When the affinity polymer was applied to purification of HSA from human serum, HSA could be purified to single-band purity according to SDS-PAGE. A thermo-response polymer was synthesized and L-thyroxin was attached to the polymer. Affinity precipitation was used to purify HSA from human serum.

  4. Affinity precipitation of human serum albumin using a thermo-response polymer with an L-thyroxin ligand

    PubMed Central

    2013-01-01

    Background Affinity precipitation has been reported as a potential technology for the purification of proteins at the early stage of downstream processing. The technology could be achieved using reversible soluble-insoluble polymers coupled with an affinity ligand to purify proteins from large volumes of dilute solution material such as fermentation broths or plasma. In this study, a thermo-response polymer was synthesized using N-methylol acrylamide, N-isopropyl acrylamide and butyl acrylate as monomers. The molecular weight of the polymer measured by the viscosity method was 3.06 × 104 Da and the lower critical solution temperature (LCST) was 28.0°C.The recovery of the polymer above the LCST was over 95.0%. Human serum albumin (HSA) is the most abundant protein in the human serum system, and it has important functions in the human body. High purity HSA is required in pharmaceuticals. Safe and efficient purification is a crucial process during HSA production. Results A thermo-response polymer was synthesized and L-thyroxin immobilized on the polymer as an affinity ligand to enable affinity precipitation of HSA. The LCST of the affinity polymer was 31.0°C and the recovery was 99.6% of its original amount after recycling three times. The optimal adsorption condition was 0.02 M Tris–HCl buffer (pH 7.0) and the HSA adsorption capacity was 14.9 mg/g polymer during affinity precipitation. Circular dichroism spectra and a ForteBio Octet system were used to analyze the interactions between the affinity polymer and HSA during adsorption and desorption. The recovery of total HSA by elution with 1.0 mol/L NaSCN was 93.6%. When the affinity polymer was applied to purification of HSA from human serum, HSA could be purified to single-band purity according to SDS-PAGE. Conclusion A thermo-response polymer was synthesized and L-thyroxin was attached to the polymer. Affinity precipitation was used to purify HSA from human serum. PMID:24341315

  5. An integrated approach to scale up the market penetration of low carbon technologies in developing countries and water scarce regions

    NASA Astrophysics Data System (ADS)

    Thompson, Michelle Angela

    Water scarcity is a global challenge that stifles social and economic growth. There is a growing concern to examine the water-energy nexus to understand the importance of applying energy and water interactions to technology. In developing countries there are many communities that live off-grid in remote region with no access to electricity or clean water. Additionally, there are developed countries that are located in regions with electricity but no access to clean water. Recent developments in renewable energy technology and energy policies have greatly reduced the costs of renewable energy making them more attractive and affordable. The purpose of this dissertation is to evaluate the main barriers to deploying renewables to non-Organization for Economic Co-operation and Development (non-OECD) countries and member countries of the Organization for Economic Co-operation and Development (OECD). This dissertation examines the potential of renewable desalination technology systems across emerging countries. The findings of this research can serve as the basis for investors interested in entering this market. The combined chapters seek to address potential problems regarding the costs, methods, and tools required for the implementation of the appropriate water purification technologies for off-grid, community scale infrastructures.

  6. Near-Zero Emissions Oxy-Combustion Flue Gas Purification Task 2: SOx/Nox/Hg Removal for High Sulfur Coal

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Nick Degenstein; Minish Shah; Doughlas Louie

    2012-05-01

    The goal of this project is to develop a near-zero emissions flue gas purification technology for existing PC (pulverized coal) power plants that are retrofitted with oxy-combustion technology. The objective of Task 2 of this project was to evaluate an alternative method of SOx, NOx and Hg removal from flue gas produced by burning high sulfur coal in oxy-combustion power plants. The goal of the program was not only to investigate a new method of flue gas purification but also to produce useful acid byproduct streams as an alternative to using a traditional FGD and SCR for flue gas processing.more » During the project two main constraints were identified that limit the ability of the process to achieve project goals. 1) Due to boiler island corrosion issues >60% of the sulfur must be removed in the boiler island with the use of an FGD. 2) A suitable method could not be found to remove NOx from the concentrated sulfuric acid product, which limits sale-ability of the acid, as well as the NOx removal efficiency of the process. Given the complexity and safety issues inherent in the cycle it is concluded that the acid product would not be directly saleable and, in this case, other flue gas purification schemes are better suited for SOx/NOx/Hg control when burning high sulfur coal, e.g. this project's Task 3 process or a traditional FGD and SCR.« less

  7. Improving the large scale purification of the HIV microbicide, griffithsin.

    PubMed

    Fuqua, Joshua L; Wanga, Valentine; Palmer, Kenneth E

    2015-02-22

    Griffithsin is a broad spectrum antiviral lectin that inhibits viral entry and maturation processes through binding clusters of oligomannose glycans on viral envelope glycoproteins. An efficient, scaleable manufacturing process for griffithsin active pharmaceutical ingredient (API) is essential for particularly cost-sensitive products such as griffithsin -based topical microbicides for HIV-1 prevention in resource poor settings. Our previously published purification method used ceramic filtration followed by two chromatography steps, resulting in a protein recovery of 30%. Our objective was to develop a scalable purification method for griffithsin expressed in Nicotiana benthamiana plants that would increase yield, reduce production costs, and simplify manufacturing techniques. Considering the future need to transfer griffithsin manufacturing technology to resource poor areas, we chose to focus modifying the purification process, paying particular attention to introducing simple, low-cost, and scalable procedures such as use of temperature, pH, ion concentration, and filtration to enhance product recovery. We achieved >99% pure griffithsin API by generating the initial green juice extract in pH 4 buffer, heating the extract to 55°C, incubating overnight with a bentonite MgCl2 mixture, and final purification with Capto™ multimodal chromatography. Griffithsin extracted with this protocol maintains activity comparable to griffithsin purified by the previously published method and we are able to recover a substantially higher yield: 88 ± 5% of griffithsin from the initial extract. The method was scaled to produce gram quantities of griffithsin with high yields, low endotoxin levels, and low purification costs maintained. The methodology developed to purify griffithsin introduces and develops multiple tools for purification of recombinant proteins from plants at an industrial scale. These tools allow for robust cost-effective production and purification of griffithsin. The methodology can be readily scaled to the bench top or industry and process components can be used for purification of additional proteins based on biophysical characteristics.

  8. An oral vaccine against candidiasis generated by a yeast molecular display system.

    PubMed

    Shibasaki, Seiji; Aoki, Wataru; Nomura, Takashi; Miyoshi, Ayuko; Tafuku, Senji; Sewaki, Tomomitsu; Ueda, Mitsuyoshi

    2013-12-01

    Enolase 1 (Eno1p) of Candida albicans is an immunodominant antigen. However, conventional technologies for preparing an injectable vaccine require purification of the antigenic protein and preparation of an adjuvant. To develop a novel type of oral vaccine against candidiasis, we generated Saccharomyces cerevisiae cells that display the Eno1p antigen on their surfaces. Oral delivery of the engineered S. cerevisiae cells prolonged survival rate of mice that were subsequently challenged with C. albicans. Given that a vaccine produced using molecular display technology avoids the need for protein purification, this oral vaccine offers a promising alternative to the use of conventional and injectable vaccines for preventing a range of infectious diseases. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  9. Hydrogen Purification Using Natural Zeolite Membranes

    NASA Technical Reports Server (NTRS)

    DelValle, William

    2003-01-01

    The School of Science at Universidad del Turabo (UT) have a long-lasting investigation plan to study the hydrogen cleaning and purification technologies. We proposed a research project for the synthesis, phase analysis and porosity characterization of zeolite based ceramic perm-selective membranes for hydrogen cleaning to support NASA's commitment to achieving a broad-based research capability focusing on aerospace-related issues. The present study will focus on technology transfer by utilizing inorganic membranes for production of ultra-clean hydrogen for application in combustion. We tested three different natural zeolite membranes (different particle size at different temperatures and time of exposure). Our results show that the membranes exposured at 900 C for 1Hr has the most higher permeation capacity, indicated that our zeolite membranes has the capacity to permeate hydrogen.

  10. Application of FTA technology to extraction of sperm DNA from mixed body fluids containing semen.

    PubMed

    Fujita, Yoshihiko; Kubo, Shin-ichi

    2006-01-01

    FTA technology is a novel method designed to simplify the collection, shipment, archiving and purification of nucleic acids from a wide variety of biological sources. In this study, we report a rapid and simple method of extracting DNA from sperm when body fluids mixed with semen were collected using FTA cards. After proteinase K digestion of the sperm and body fluid mixture, the washed pellet suspension as the sperm fraction and the concentrated supernatant as the epithelial cell fraction were respectively applied to FTA cards containing DTT. The FTA cards were dried, then directly added to a polymerase chain reaction (PCR) mix and processed by PCR. The time required from separation of the mixed fluid into sperm and epithelial origin DNA extractions was only about 2.5-3h. Furthermore, the procedure was extremely simple. It is considered that our designed DNA extraction procedure using an FTA card is available for application to routine work.

  11. Field Application of the Micro Biological Survey Method for the Assessment of the Microbiological Safety of Different Water Sources in Horn of Africa and the Evaluation of the Effectiveness of Moringa Oleifera in Drinking Water Purification

    PubMed Central

    Losito, Francesca; Arienzo, Alyexandra; Somma, Daniela; Murgia, Lorenza; Stalio, Ottavia; Zuppi, Paolo; Rossi, Elisabetta; Antonini, Giovanni

    2017-01-01

    Water monitoring requires expensive instrumentations and skilled technicians. In developing Countries as Africa, the severe economic restrictions and lack of technology make water safety monitoring approaches applied in developed Countries, still not sustainable. The need to develop new methods that are suitable, affordable, and sustainable in the African context is urgent. The simple, economic and rapid Micro Biological Survey (MBS) method does not require an equipped laboratory nor special instruments and skilled technicians, but it can be very useful for routine water analysis. The aim of this work was the application of the MBS method to evaluate the microbiological safety of different water sources and the effectiveness of different drinking water treatments in the Horn of Africa. The obtained results have proved that this method could be very helpful to monitor water safety before and after various purification treatments, with the aim to control waterborne diseases especially in developing Countries, whose population is the most exposed to these diseases. In addition, it has been proved that Moringa oleifera water treatment is ineffective in decreasing bacterial load of Eritrea water samples. PMID:28748063

  12. Aqueous two-phase partition applied to the isolation of plasma membranes and Golgi apparatus from cultured mammalian cells.

    PubMed

    Morré, D M; Morre, D J

    2000-06-23

    Partitioning in dextran-poly(ethylene)glycol (PEG) aqueous-aqueous phase systems represents a mature technology with many applications to separations of cells and to the preparation of membranes from mammalian cells. Most applications to membrane isolation and purification have focused on plasma membranes, plasma membrane domains and separation of right side-out and inside-out plasma membrane vesicles. The method exploits a combination of membrane properties, including charge and hydrophobicity. Purification is based upon differential distributions of the constituents in a sample between the two principal compartments of the two phases (upper and lower) and at the interface. The order of affinity of animal cell membranes for the upper phase is: endoplasmic reticulum

  13. Field Application of the Micro Biological Survey Method for the Assessment of the Microbiological Safety of Different Water Sources in Horn of Africa and the Evaluation of the Effectiveness of Moringa Oleifera in Drinking Water Purification.

    PubMed

    Losito, Francesca; Arienzo, Alyexandra; Somma, Daniela; Murgia, Lorenza; Stalio, Ottavia; Zuppi, Paolo; Rossi, Elisabetta; Antonini, Giovanni

    2017-06-23

    Water monitoring requires expensive instrumentations and skilled technicians. In developing Countries as Africa, the severe economic restrictions and lack of technology make water safety monitoring approaches applied in developed Countries, still not sustainable. The need to develop new methods that are suitable, affordable, and sustainable in the African context is urgent. The simple, economic and rapid Micro Biological Survey (MBS) method does not require an equipped laboratory nor special instruments and skilled technicians, but it can be very useful for routine water analysis. The aim of this work was the application of the MBS method to evaluate the microbiological safety of different water sources and the effectiveness of different drinking water treatments in the Horn of Africa. The obtained results have proved that this method could be very helpful to monitor water safety before and after various purification treatments, with the aim to control waterborne diseases especially in developing Countries, whose population is the most exposed to these diseases. In addition, it has been proved that Moringa oleifera water treatment is ineffective in decreasing bacterial load of Eritrea water samples.

  14. Aqueous two-phase partition applied to the isolation of plasma membranes and Golgi apparatus from cultured mammalian cells

    NASA Technical Reports Server (NTRS)

    Morre, D. M.; Morre, D. J.

    2000-01-01

    Partitioning in dextran-poly(ethylene)glycol (PEG) aqueous-aqueous phase systems represents a mature technology with many applications to separations of cells and to the preparation of membranes from mammalian cells. Most applications to membrane isolation and purification have focused on plasma membranes, plasma membrane domains and separation of right side-out and inside-out plasma membrane vesicles. The method exploits a combination of membrane properties, including charge and hydrophobicity. Purification is based upon differential distributions of the constituents in a sample between the two principal compartments of the two phases (upper and lower) and at the interface. The order of affinity of animal cell membranes for the upper phase is: endoplasmic reticulum

  15. Chemical purification of lanthanides for low-background experiments

    NASA Astrophysics Data System (ADS)

    Boiko, R. S.

    2017-10-01

    There are many potentially active isotopes among the lanthanide elements which are possible to use for low-background experiments to search for double β decay, dark matter, to investigate rare α and β decays. These kind of experiments require very low level of radioactive contamination, but commercially available compounds of lanthanides are always contamined by uranium, thorium, radium, potassium, etc. A simple chemical method based on liquid-liquid extraction has been applied for the purification of CeO2, Nd2O3 and Gd˙2O˙3 from radioactive traces. Detailed schemes of purification procedure are described. Measurements by using HPGe spectrometry demonstrate high efficiency in K, Ra, Th, U contaminations reduction on at least one order of magnitude.

  16. Water Recovery from Brine in the Short and Long Term: A KSC Approach

    NASA Technical Reports Server (NTRS)

    Lunn, Griffin; Melendez, Orlando; Anthony, Steve

    2014-01-01

    KSC has spent many years researching Hollow Fiber Membrane Bioreactors as well as research encompassing:Alternate ammonia removal/Advanced oxidation. Brine purification technologies KSC-ISRU has built an electrolysis cell for the removal of acids in ISRU mining brines. Our goal is to combine all such technologies.

  17. Emerging Materials Technologies That Matter to Manufacturers

    NASA Technical Reports Server (NTRS)

    Misra, Ajay K.

    2015-01-01

    A brief overview of emerging materials technologies. Exploring the weight reduction benefit of replacing Carbon Fiber with Carbon Nanotube (CNT) in Polymer Composites. Review of the benign purification method developed for CNT sheets. The future of manufacturing will include the integration of computational material design and big data analytics, along with Nanomaterials as building blocks.

  18. Application of Moving Bed Biofilm Reactor (MBBR) and Integrated Fixed Activated Sludge (IFAS) for Biological River Water Purification System: A Short Review

    NASA Astrophysics Data System (ADS)

    Lariyah, M. S.; Mohiyaden, H. A.; Hayder, G.; Hayder, G.; Hussein, A.; Basri, H.; Sabri, A. F.; Noh, MN

    2016-03-01

    This review paper present the MBBR and IFAS technology for urban river water purification including both conventional methods and new emerging technologies. The aim of this paper is to present the MBBR and IFAS technology as an alternative and successful method for treating different kinds of effluents under different condition. There are still current treatment technologies being researched and the outcomes maybe available in a while. The review also includes many relevant researches carried out at the laboratory and pilot scales. This review covers the important processes on MBBR and IFAS basic treatment process, affecting of carrier type and influent types. However, the research concluded so far are compiled herein and reported for the first time to acquire a better perspective and insight on the subject with a view of meeting the news approach. The research concluded so far are compiled herein and reported for the first time to acquire a better perspective and insight on the subject with a view of meeting the news approach. To this end, the most feasible technology could be the combination of advanced biological process (bioreactor systems) including MBBR and IFAS system.

  19. Application of NASA's Advanced Life Support Technologies for Waste Treatment, Water Purification and Recycle, and Food Production in Polar Regions

    NASA Technical Reports Server (NTRS)

    Bubenheim, David L.; Lewis, Carol E.; Covington, M. Alan (Technical Monitor)

    1995-01-01

    NASA's advanced life support technologies are being combined with Arctic science and engineering knowledge to address the unique needs of the remote communities of Alaska through the Advanced Life Systems for Extreme Environments (ALSEE) project. ALSEE is a collaborative effort involving NASA, the State of Alaska, the University of Alaska, the North Slope Borough of Alaska, and the National Science Foundation (NSF). The focus is a major issue in the state of Alaska and other areas of the Circumpolar North, the health and welfare of its people, their lives and the subsistence lifestyle in remote communities, economic opportunity, and care for the environment. The project primarily provides treatment and reduction of waste, purification and recycling of water. and production of food. A testbed is being established to demonstrate the technologies which will enable safe, healthy, and autonomous function of remote communities and to establish the base for commercial development of the resulting technology into new industries. The challenge is to implement the technological capabilities in a manner compatible with the social and economic structures of the native communities, the state, and the commercial sector. Additional information is contained in the original extended abstract.

  20. Application of alkaline solid residue of electric arc furnace dust for neutralization/purification of electroplating wastewaters.

    PubMed

    Elez, Loris; Orescanin, Visnja; Sofilic, Tahir; Mikulic, Nenad; Ruk, Damir

    2008-10-01

    The purpose of this work was development of an appropriate procedure for the neutralization/purification of electroplating wastewater (EWW) with alkaline solid residue (ASR) by-product of the alkaline extraction of zinc and lead from electric arc furnace dust (EAFD). Removal efficiency of ASR at optimum purification conditions (pH 8 and mixing time; 20 minutes) for the elements Pb, Cr (VI), Cr (III), Fe, Ni, Cu and Zn were 94.92%, 97.58%, 99.59%, 99.48%, 97.25% and 99.97%, respectively. The concentrations of all elements in the purified wastewater were significantly lower in relation to the upper permissible limit for wastewaters suitable for discharge into the environment. The remaining waste mud was regenerated in the strong alkaline medium and successfully applied once again for the neutralization/purification of EWW. Removal efficiencies of heavy metals accomplished with regenerated waste mud were comparable to these achieved by original ASR. Elemental concentrations in the leachates of the waste mud were in accordance with regulated values.

  1. Microgravity

    NASA Image and Video Library

    2004-04-15

    Ecomaster, an affiliate of BioServe Space Technologies, this PentaPure technology has been used to purify water for our nation's Space Shuttle missions since 1981. WTC-Ecomaster of Mirneapolis, Minnesota manufactures water purification systems under the brand name PentaPure (TM). BioServe researcher Dr. George Marchin, of Kansas State University, first demonstrated the superiority of this technology and licensed it to WTC. Marchin continues to perform microgravity research in the development of new technologies for the benefit of life on Earth.

  2. Efficient, ultra-high-affinity chromatography in a one-step purification of complex proteins

    PubMed Central

    Vassylyeva, Marina N.; Klyuyev, Sergiy; Vassylyev, Alexey D.; Wesson, Hunter; Zhang, Zhuo; Renfrow, Matthew B.; Wang, Hengbin; Higgins, N. Patrick; Chow, Louise T.; Vassylyev, Dmitry G.

    2017-01-01

    Protein purification is an essential primary step in numerous biological studies. It is particularly significant for the rapidly emerging high-throughput fields, such as proteomics, interactomics, and drug discovery. Moreover, purifications for structural and industrial applications should meet the requirement of high yield, high purity, and high activity (HHH). It is, therefore, highly desirable to have an efficient purification system with a potential to meet the HHH benchmark in a single step. Here, we report a chromatographic technology based on the ultra-high-affinity (Kd ∼ 10−14–10−17 M) complex between the Colicin E7 DNase (CE7) and its inhibitor, Immunity protein 7 (Im7). For this application, we mutated CE7 to create a CL7 tag, which retained the full binding affinity to Im7 but was inactivated as a DNase. To achieve high capacity, we developed a protocol for a large-scale production and highly specific immobilization of Im7 to a solid support. We demonstrated its utility with one-step HHH purification of a wide range of traditionally challenging biological molecules, including eukaryotic, membrane, toxic, and multisubunit DNA/RNA-binding proteins. The system is simple, reusable, and also applicable to pulldown and kinetic activity/binding assays. PMID:28607052

  3. [Recombinant granulocyte-colony stimulating factor (filgrastim): optimization of conditions of isolation and purification from inclusion body].

    PubMed

    Kononova, N V; Iakovlev, A V; Zhuravko, A M; Pankeev, N N; Minaev, S V; Bobruskin, A I; Mart'ianov, V A

    2014-01-01

    We developed a unified process platform for two recombinant human GCSF medicines--one with the non-prolonged and the other with prolonged action. This unified technology led to a simpler and cheaper production while introduction of the additional pegylation stage to the technological line eased obtaining of the medicines with different action and allowed to standardize technological process documenting according to GMP requirements.

  4. Solid Phase Extraction (SPE) for Biodiesel Processing and Analysis

    DTIC Science & Technology

    2017-12-13

    1 METHODS ...sources. There are several methods than can be applied to development of separation techniques that may replace necessary water wash steps in...biodiesel refinement. Unfortunately, the most common methods are poorly suited or face high costs when applied to diesel purification. Distillation is

  5. Methods of cell purification: a critical juncture for laboratory research and translational science.

    PubMed

    Amos, Peter J; Cagavi Bozkulak, Esra; Qyang, Yibing

    2012-01-01

    Research in cell biology and the development of translational technologies are driven by competition, public expectations, and regulatory oversight, putting these fields at a critical juncture. Success in these fields is quickly becoming dependent on the ability of researchers to identify and isolate specific cell populations from heterogeneous mixtures accurately and efficiently. Many methods for cell purification have been developed, and each has advantages and disadvantages that must be considered in light of the intended application. Current cell separation strategies make use of surface proteins, genetic expression, and physics to isolate specific cells by phenotypic traits. Cell purification is also dependent on the cellular reagents available for use and the intended application, as these factors may preclude certain mechanisms used in the processes of labeling and sorting cells. Copyright © 2011 S. Karger AG, Basel.

  6. Adsorption processes in spacecraft environmental control and life support systems

    NASA Technical Reports Server (NTRS)

    DallBauman, L. A.; Finn, J. E.

    1999-01-01

    The environmental control and life support system on a spacecraft maintains a safe and comfortable environment in which the crew can live and work by supplying oxygen and water and by removing carbon dioxide, water vapor, and trace contaminants from cabin air. Although open-loop systems have been used successfully in the past for short-duration missions, the economics of current and future long-duration missions in space will make nearly complete recycling of air and water imperative. A variety of operations will be necessary to achieve the goal of nearly complete recycling. These include separation and reduction of carbon dioxide, removal of trace gas-phase contaminants, recovery and purification of humidity condensate, purification and polishing of wastewater streams, and others. Several of these can be performed totally or in part by adsorption processes. These processes are good candidates to perform separations and purifications in space due to their gravity independence, high reliability, relative high energy efficiency, design flexibility, technological maturity, and regenerative nature. For these reasons, adsorption has historically played a key role in life support on U.S. and Russian piloted spacecraft. Among the life support applications that can be achieved through use of adsorption technology are removal of trace contaminants and carbon dioxide from cabin air and recovery of potable water from waste streams. In each of these cases adsorption technology has been selected for use onboard the International Space Station. The requirements, science, and hardware for these applications are discussed. Human space exploration may eventually lead to construction of planetary habitats. These habitats may provide additional opportunities for use of adsorption processes, such as control of greenhouse gas composition, and may have different resources available to them, such as gases present in the planetary atmosphere. Separation and purification processes based on adsorption can be expected to continue to fulfill environmental control and life support needs on future missions.

  7. Adsorption processes in spacecraft environmental control and life support systems.

    PubMed

    DallBauman, L A; Finn, J E

    1999-01-01

    The environmental control and life support system on a spacecraft maintains a safe and comfortable environment in which the crew can live and work by supplying oxygen and water and by removing carbon dioxide, water vapor, and trace contaminants from cabin air. Although open-loop systems have been used successfully in the past for short-duration missions, the economics of current and future long-duration missions in space will make nearly complete recycling of air and water imperative. A variety of operations will be necessary to achieve the goal of nearly complete recycling. These include separation and reduction of carbon dioxide, removal of trace gas-phase contaminants, recovery and purification of humidity condensate, purification and polishing of wastewater streams, and others. Several of these can be performed totally or in part by adsorption processes. These processes are good candidates to perform separations and purifications in space due to their gravity independence, high reliability, relative high energy efficiency, design flexibility, technological maturity, and regenerative nature. For these reasons, adsorption has historically played a key role in life support on U.S. and Russian piloted spacecraft. Among the life support applications that can be achieved through use of adsorption technology are removal of trace contaminants and carbon dioxide from cabin air and recovery of potable water from waste streams. In each of these cases adsorption technology has been selected for use onboard the International Space Station. The requirements, science, and hardware for these applications are discussed. Human space exploration may eventually lead to construction of planetary habitats. These habitats may provide additional opportunities for use of adsorption processes, such as control of greenhouse gas composition, and may have different resources available to them, such as gases present in the planetary atmosphere. Separation and purification processes based on adsorption can be expected to continue to fulfill environmental control and life support needs on future missions.

  8. Adsorption Processes in Spacecraft Environmental Control and Life Support Systems

    NASA Technical Reports Server (NTRS)

    Bauman, Liese Dall; Finn, John E.; Kliss, Mark (Technical Monitor)

    1998-01-01

    The environmental control and life support system on a spacecraft must maintain a safe and comfortable environment in which the crew can live and work. The system's functions include supplying the crew with oxygen and water as well as removing carbon dioxide, water vapor, and trace contaminants from cabin air. Although open-loop systems have been used in the past, logistics and safety factors of current and future missions in space make near-complete recycling of the cabin's air and water imperative. The recycling process may include separation and reduction of carbon dioxide, removal of trace gas-phase contaminants, recovery and purification of humidity condensate, purification and polishing of wastewater streams, and other processes. Several of these operations can be performed totally or in part by adsorption processes. These processes are frequently good candidates to perform separations and purifications in space due to their gravity independence, high reliability, relatively high energy efficiency, design flexibility, technological maturity, and regenerability. For these reasons, adsorption has historically played a key role in life support on U.S. and Russian piloted spacecraft. This article focuses on three current spacecraft life support applications that often use adsorption technology: gas-phase trace contaminant control, carbon dioxide removal from cabin air, and potable water recovery from waste streams. In each application, adsorption technology has been selected for use on the International Space Station. The requirements, science, and hardware for each of these applications are discussed. Eventually, human space exploration may lead to construction of planetary habitats. These habitats may provide additional opportunities for use of adsorption processes, such as control of greenhouse gas composition, and may have different requirements and resources available to them, such as gases present in the planetary atmosphere. Adsorption separation and purification processes can be expected to continue to fulfill environmental control and life support needs on future missions.

  9. The Toxic Truth About Carbon Nanotubes in Water Purification: a Perspective View.

    PubMed

    Das, Rasel; Leo, Bey Fen; Murphy, Finbarr

    2018-06-18

    Without nanosafety guidelines, the long-term sustainability of carbon nanotubes (CNTs) for water purifications is questionable. Current risk measurements of CNTs are overshadowed by uncertainties. New risks associated with CNTs are evolving through different waste water purification routes, and there are knowledge gaps in the risk assessment of CNTs based on their physical properties. Although scientific efforts to design risk estimates are evolving, there remains a paucity of knowledge on the unknown health risks of CNTs. The absence of universal CNT safety guidelines is a specific hindrance. In this paper, we close these gaps and suggested several new risk analysis roots and framework extrapolations from CNT-based water purification technologies. We propose a CNT safety clock that will help assess risk appraisal and management. We suggest that this could form the basis of an acceptable CNT safety guideline. We pay particular emphasis on measuring risks based on CNT physico-chemical properties such as diameter, length, aspect ratio, type, charge, hydrophobicity, functionalities and so on which determine CNT behaviour in waste water treatment plants and subsequent release into the environment.

  10. SNO+ Scintillator Purification and Assay

    NASA Astrophysics Data System (ADS)

    Ford, R.; Chen, M.; Chkvorets, O.; Hallman, D.; Vázquez-Jáuregui, E.

    2011-04-01

    We describe the R&D on the scintillator purification and assay methods and technology for the SNO+ neutrino and double-beta decay experiment. The SNO+ experiment is a replacement of the SNO heavy water with liquid scintillator comprised of 2 g/L PPO in linear alkylbenzene (LAB). During filling the LAB will be transported underground by rail car and purified by multi-stage distillation and steam stripping at a flow rate of 19 LPM. While the detector is operational the scintillator can be recirculated at 150 LPM (full detector volume in 4 days) to provide repurification as necessary by either water extraction (for Ra, K, Bi) or by functional metal scavenger columns (for Pb, Ra, Bi, Ac, Th) followed by steam stripping to remove noble gases and oxygen (Rn, O2, Kr, Ar). The metal scavenger columns also provide a method for scintillator assay for ex-situ measurement of the U and Th chain radioactivity. We have developed "natural" radioactive spikes of Pb and Ra in LAB and use these for purification testing. Lastly, we present the planned operating modes and purification strategies and the plant specifications and design.

  11. Water Treatment: Can You Purify Water for Drinking?

    ERIC Educational Resources Information Center

    Harris, Mary E.

    1996-01-01

    Presents a three-day mini unit on purification of drinking water that uses the learning cycle approach. Demonstrates the typical technology that water companies use to provide high-quality drinking water. (JRH)

  12. Potential technology transfers of research on low-temperature carbon monoxide-oxygen recombination catalysts

    NASA Technical Reports Server (NTRS)

    Poziomek, Edward J.

    1990-01-01

    Results from research on catalytic recombination of CO-O2 for stable closed-cycle operation of CO2 lasers hold much promise for a variety of technology transfer. Expansion of CO2 laser remote sensing applications toward chemical detection and pollution monitoring would certainly be expected. However, the catalysts themselves may be especially effective in low-temperature oxidation of a number of chemicals in addition to CO. It is therefore of interest to compare the CO-O2 catalysts with chemical systems designed for chemical sensing, air purification and process catalysis. Success in understanding the catalytic mechanisms of the recombination of CO-O2 could help to shed light on how catalyst systems operate. New directions in low-temperature oxidation catalysts, coatings for chemical sensors and sorbents for air purification could well emerge.

  13. IGIV: contents, properties, and methods of industrial production--evolving closer to a more physiologic product.

    PubMed

    Martin, Turf D

    2006-04-01

    Is the process the product? Immune globulin intravenous (IGIV) is not manufactured, but is purified (fractionated) from human plasma. Machines can only damage what Mother Nature makes; they cannot improve it. Therefore, fractionators of biologic molecules must strive to ensure what is taken from a human body is exactly the same when it is returned to the human body for optimal tolerability and safety. The processes of purification have the potential to adversely affect the product. Four primary purification processes exist for commercial IGIV. The Cohn-Oncley process is 1940s technology, which has been modified through the decades, but the basic process remains unchanged. The Kistler-Nitschmann process was developed in the 1950s by the Central Laboratory of the Swiss Red Cross (ZLB, today known as ZLB-Behring, a subsidiary of CSL Limited). Various attempts have been made to utilize chromatography as the sole separation technology without much success. Most recently, Bayer HealthCare (Talecris Biotherapeutics acquired the contributed assets of the worldwide plasma business of Bayer Biological Products and became operational April 1, 2005; all plasma-based products, including Gamunex, Prolastin, the hyperimmune line (Fraction II), Plasbumin (Bayer Albumin), Koate DVI, and Thrombate III were included) introduced a new product into the United States and Canada that utilizes caprylate and chromatography for high purity, better yields, and integration of safety and efficacy. This is the first new IGIV purification technology in over 20 years.

  14. Recent Methods for Purification and Structure Determination of Oligonucleotides.

    PubMed

    Zhang, Qiulong; Lv, Huanhuan; Wang, Lili; Chen, Man; Li, Fangfei; Liang, Chao; Yu, Yuanyuan; Jiang, Feng; Lu, Aiping; Zhang, Ge

    2016-12-18

    Aptamers are single-stranded DNA or RNA oligonucleotides that can interact with target molecules through specific three-dimensional structures. The excellent features, such as high specificity and affinity for target proteins, small size, chemical stability, low immunogenicity, facile chemical synthesis, versatility in structural design and engineering, and accessible for site-specific modifications with functional moieties, make aptamers attractive molecules in the fields of clinical diagnostics and biopharmaceutical therapeutics. However, difficulties in purification and structural identification of aptamers remain a major impediment to their broad clinical application. In this mini-review, we present the recently attractive developments regarding the purification and identification of aptamers. We also discuss the advantages, limitations, and prospects for the major methods applied in purifying and identifying aptamers, which could facilitate the application of aptamers.

  15. Hydrogen Purification and Recycling for an Integrated Oxygen Recovery System Architecture

    NASA Technical Reports Server (NTRS)

    Abney, Morgan B.; Greenwood, Zachary; Wall, Terry; Miller, Lee; Wheeler, Ray

    2016-01-01

    The United States Atmosphere Revitalization life support system on the International Space Station (ISS) performs several services for the crew including oxygen generation, trace contaminant control, carbon dioxide (CO2) removal, and oxygen recovery. Oxygen recovery is performed using a Sabatier reactor developed by Hamilton Sundstrand, wherein CO2 is reduced with hydrogen in a catalytic reactor to produce methane and water. The water product is purified in the Water Purification Assembly and recycled to the Oxygen Generation Assembly (OGA) to provide O2 to the crew. This architecture results in a theoretical maximum oxygen recovery from CO2 of approximately 54% due to the loss of reactant hydrogen in Sabatier-produced methane that is currently vented outside of ISS. Plasma Methane Pyrolysis technology (PPA), developed by Umpqua Research Company, provides the capability to further close the Atmosphere Revitalization oxygen loop by recovering hydrogen from Sabatier-produced methane. A key aspect of this technology approach is to purify the hydrogen from the PPA product stream which includes acetylene, unreacted methane and byproduct water and carbon monoxide. In 2015, four sub-scale hydrogen separation systems were delivered to NASA for evaluation. These included two electrolysis single-cell hydrogen purification cell stacks developed by Sustainable Innovations, LLC, a sorbent-based hydrogen purification unit using microwave power for sorbent regeneration developed by Umpqua Research Company, and a LaNi4.6Sn0.4 metal hydride produced by Hydrogen Consultants, Inc. Here we report the results of these evaluations, discuss potential architecture options, and propose future work.

  16. Hydrogen Purification and Recycling for an Integrated Oxygen Recovery System Architecture

    NASA Technical Reports Server (NTRS)

    Abney, Morgan B.; Greenwood, Zachary; Wall, Terry; Nur, Mononita; Wheeler, Richard R., Jr.; Preston, Joshua; Molter, Trent

    2016-01-01

    The United States Atmosphere Revitalization life support system on the International Space Station (ISS) performs several services for the crew including oxygen generation, trace contaminant control, carbon dioxide (CO2) removal, and oxygen recovery. Oxygen recovery is performed using a Sabatier reactor developed by Hamilton Sundstrand, wherein CO2 is reduced with hydrogen in a catalytic reactor to produce methane and water. The water product is purified in the Water Purification Assembly and recycled to the Oxygen Generation Assembly (OGA) to provide O2 to the crew. This architecture results in a theoretical maximum oxygen recovery from CO2 of approx.54% due to the loss of reactant hydrogen in Sabatier-produced methane that is currently vented outside of ISS. Plasma Pyrolysis Assembly (PPA) technology, developed by Umpqua Research Company, provides the capability to further close the Atmosphere Revitalization oxygen loop by recovering hydrogen from Sabatier-produced methane. A key aspect of this technology approach is the need to purify the hydrogen from the PPA product stream which includes acetylene, unreacted methane and byproduct water and carbon monoxide. In 2015, four sub-scale hydrogen separation systems were delivered to NASA for evaluation. These included two electrolysis single-cell hydrogen purification cell stacks developed by Sustainable Innovations, LLC, a sorbent-based hydrogen purification unit using microwave power for sorbent regeneration developed by Umpqua Research Company, and a LaNi4.6Sn0.4 metal hydride produced by Hydrogen Consultants, Inc. Here we report the results of these evaluations to-date, discuss potential architecture options, and propose future work.

  17. On-Line Ion Exchange Liquid Chromatography as a Process Analytical Technology for Monoclonal Antibody Characterization in Continuous Bioprocessing.

    PubMed

    Patel, Bhumit A; Pinto, Nuno D S; Gospodarek, Adrian; Kilgore, Bruce; Goswami, Kudrat; Napoli, William N; Desai, Jayesh; Heo, Jun H; Panzera, Dominick; Pollard, David; Richardson, Daisy; Brower, Mark; Richardson, Douglas D

    2017-11-07

    Combining process analytical technology (PAT) with continuous production provides a powerful tool to observe and control monoclonal antibody (mAb) fermentation and purification processes. This work demonstrates on-line liquid chromatography (on-line LC) as a PAT tool for monitoring a continuous biologics process and forced degradation studies. Specifically, this work focused on ion exchange chromatography (IEX), which is a critical separation technique to detect charge variants. Product-related impurities, including charge variants, that impact function are classified as critical quality attributes (CQAs). First, we confirmed no significant differences were observed in the charge heterogeneity profile of a mAb through both at-line and on-line sampling and that the on-line method has the ability to rapidly detect changes in protein quality over time. The robustness and versatility of the PAT methods were tested by sampling from two purification locations in a continuous mAb process. The PAT IEX methods used with on-line LC were a weak cation exchange (WCX) separation and a newly developed shorter strong cation exchange (SCX) assay. Both methods provided similar results with the distribution of percent acidic, main, and basic species remaining unchanged over a 2 week period. Second, a forced degradation study showed an increase in acidic species and a decrease in basic species when sampled on-line over 7 days. These applications further strengthen the use of on-line LC to monitor CQAs of a mAb continuously with various PAT IEX analytical methods. Implementation of on-line IEX will enable faster decision making during process development and could potentially be applied to control in biomanufacturing.

  18. Development of a Microwave Regenerative Sorbent-Based Hydrogen Purifier

    NASA Technical Reports Server (NTRS)

    Wheeler, Richard R., Jr.; Dewberry, Ross H.; McCurry, Bryan D.; Abney, Morgan B.; Greenwood, Zachary W.

    2016-01-01

    This paper describes the design and fabrication of a Microwave Regenerative Sorbent-based Hydrogen Purifier (MRSHP). This unique microwave powered technology was developed for the purification of a hydrogen stream produced by the Plasma Pyrolysis Assembly (PPA). The PPA is a hydrogen recovery (from methane) post processor for NASA's Sabatier-based carbon dioxide reduction process. Embodied in the Carbon dioxide Reduction Assembly (CRA), currently aboard the International Space Station (ISS), the Sabatier reaction employs hydrogen to catalytically recover oxygen, in the form of water, from respiratory carbon dioxide produced by the crew. This same approach is base-lined for future service in the Air Revitalization system on extended missions into deep space where resupply is not practical. Accordingly, manned exploration to Mars may only become feasible with further closure of the air loop as afforded by the greater hydrogen recovery permitted by the PPA with subsequent hydrogen purification. By utilizing the well-known high sorbate loading capacity of molecular sieve 13x, coupled with microwave dielectric heating phenomenon, MRSHP technology is employed as a regenerative filter for a contaminated hydrogen gas stream. By design, freshly regenerated molecular sieve 13x contained in the MRSHP will remove contaminants from the effluent of a 1-CM scale PPA for several hours prior to breakthrough. By reversing flow and pulling a relative vacuum the MRSHP prototype then uses 2.45 GHz microwave power, applied through a novel coaxial antenna array, to rapidly heat the sorbent bed and drive off the contaminants in a short duration vacuum/thermal contaminant desorption step. Finally, following rapid cooling via room temperature cold plates, the MRSHP is again ready to serve as a hydrogen filter.

  19. Research and preparation of ultra purity silicon tetrachloride

    NASA Astrophysics Data System (ADS)

    Wan, Ye; Zhao, Xiong; Yan, Dazhou; Yang, Dian; Li, Yunhao; Guo, Shuhu

    2017-10-01

    This article demonstrated a technology for producing ultra-purity silicon tetrachloride, which using the high purity SiCl4 as raw material through the method of combination ray reaction with purification. This technology could remove metal impurities and compounds impurities contained hydrogen effectively. The purity of product prepared by this technology can reach at 99.9999%, content of metal impurities can be low at 0.3PPb, meeting the requirement of industry easily. This technology has the advantages of simple process, continuous operation, and stable performance.

  20. Force Projection Technology Overview

    DTIC Science & Technology

    2011-08-12

    Technologies • Fuel Efficient Powertrain Lubricant • Nanotechnology for Fuels and Lubes • Water from Air • Water Reuse • In-line Water Monitoring...purification systems with new pretreatment, desalination and post treatment technologies. Payoff: • Reduces the logistical footprint associated with water...FY11 FY12 FY13 FY14 FY15 FY16 FY17 •Water From Air •Water Quality Monitoring •Water Reuse •Pre and Post Treatment • Desalination 6 5 5

  1. CLEAN PRODUCTION WITH MEMBRANE TECHNOLOGY

    EPA Science Inventory

    Membrane processes, because of unique, specialized, and cost-effective applications, have the potential of playing a significant role in preventing pollution from occurring in manufacturing plans. Opportunities are seen in resource recovery, species purification, and energy sav...

  2. Tannin extracts from immature fruits of Terminalia chebula Fructus Retz. promote cutaneous wound healing in rats

    PubMed Central

    2011-01-01

    Background Tannins extracted from immature fruits of Terminalia chebula Fructus Retz. are considered as effective components promoting the process of wound healing. The objective of this study is to explore the optimal extraction and purification technology (OEPT) of tannins, while studying the use of this drug in the treatment of a cutaneous wound of rat as well as its antibacterial effects. Methods The content of tannin extracts was measured by the casein method, and antibacterial ability was studied by the micro-dilution method in vitro. In wound healing experiment, animals in group Ⅰ, Ⅱ and Ⅲ were treated with vaseline ointment, tannin extracts (tannin content: 81%) and erythromycin ointment, respectively (5 mg of ointment were applied on each wound). To evaluate the process of wound healing, selected pharmacological and biochemical parameters were applied. Results After optimal extraction and purification, content of tannin extracts was increased to 81%. Tannin extracts showed the inhibition of Staphylococcus aureus and Klebsiella Pneumonia in vitro. After excision of wounds, on days 7 and 10, the percent of wound contraction of group Ⅱ was higher than that of group Ⅰ. After being hurt with wounds, on days 3, 7, and 10, the wound healing quality of group Ⅱ was found to be better than that of group Ⅰ in terms of granulation formation and collagen organization. After wound creation, on day 3, the vascular endothelial growth factor expression of group Ⅱ was higher than that of group Ⅰ. Conclusion The results suggest that tannin extracts from dried immature fruits of Terminalia chebula Fructus Retz. can promote cutaneous wound healing in rats, probably resulting from a powerful anti-bacterial and angiogenic activity of the extracts. PMID:21982053

  3. Combustion water purification techniques influence on OBT analysing using liquid scintillation counting method

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Varlam, C.; Vagner, I.; Faurescu, I.

    In order to determine organically bound tritium (OBT) from environmental samples, these must be converted into water, measurable by liquid scintillation counting (LSC). For this purpose we conducted some experiments to determine OBT level of a grass sample collected from an uncontaminated area. The studied grass sample was combusted in a Parr bomb. However usual interfering phenomena were identified: color or chemical quench, chemiluminescence, overlap over tritium spectrum because of other radionuclides presence as impurities ({sup 14}C from organically compounds, {sup 36}Cl as chloride and free chlorine, {sup 40}K as potassium cations) and emulsion separation. So the purification of themore » combustion water before scintillation counting appeared to be essential. 5 purification methods were tested: distillation with chemical treatment (Na{sub 2}O{sub 2} and KMnO{sub 4}), lyophilization, chemical treatment (Na{sub 2}O{sub 2} and KMnO{sub 4}) followed by lyophilization, azeotropic distillation with toluene and treatment with a volcanic tuff followed by lyophilization. After the purification step each sample was measured and the OBT measured concentration, together with physico-chemical analysis of the water analyzed, revealed that the most efficient method applied for purification of the combustion water was the method using chemical treatment followed by lyophilization.« less

  4. Reversed-phase ion-pair liquid chromatography method for purification of duplex DNA with single base pair resolution

    PubMed Central

    Wysoczynski, Christina L.; Roemer, Sarah C.; Dostal, Vishantie; Barkley, Robert M.; Churchill, Mair E. A.; Malarkey, Christopher S.

    2013-01-01

    Obtaining quantities of highly pure duplex DNA is a bottleneck in the biophysical analysis of protein–DNA complexes. In traditional DNA purification methods, the individual cognate DNA strands are purified separately before annealing to form DNA duplexes. This approach works well for palindromic sequences, in which top and bottom strands are identical and duplex formation is typically complete. However, in cases where the DNA is non-palindromic, excess of single-stranded DNA must be removed through additional purification steps to prevent it from interfering in further experiments. Here we describe and apply a novel reversed-phase ion-pair liquid chromatography purification method for double-stranded DNA ranging in lengths from 17 to 51 bp. Both palindromic and non-palindromic DNA can be readily purified. This method has the unique ability to separate blunt double-stranded DNA from pre-attenuated (n-1, n-2, etc) synthesis products, and from DNA duplexes with single base pair overhangs. Additionally, palindromic DNA sequences with only minor differences in the central spacer sequence of the DNA can be separated, and the purified DNA is suitable for co-crystallization of protein–DNA complexes. Thus, double-stranded ion-pair liquid chromatography is a useful approach for duplex DNA purification for many applications. PMID:24013567

  5. Case studies on the physical-chemical parameters' variation during three different purification approaches destined to treat wastewaters from food industry.

    PubMed

    Ghimpusan, Marieta; Nechifor, Gheorghe; Nechifor, Aurelia-Cristina; Dima, Stefan-Ovidiu; Passeri, Piero

    2017-12-01

    The paper presents a set of three interconnected case studies on the depuration of food processing wastewaters by using aeration & ozonation and two types of hollow-fiber membrane bioreactor (MBR) approaches. A secondary and more extensive objective derived from the first one is to draw a clearer, broader frame on the variation of physical-chemical parameters during the purification of wastewaters from food industry through different operating modes with the aim of improving the management of water purification process. Chemical oxygen demand (COD), pH, mixed liquor suspended solids (MLSS), total nitrogen, specific nitrogen (NH 4 + , NO 2 - , NO 3 - ) total phosphorous, and total surfactants were the measured parameters, and their influence was discussed in order to establish the best operating mode to achieve the purification performances. The integrated air-ozone aeration process applied in the second operating mode lead to a COD decrease by up to 90%, compared to only 75% obtained in a conventional biological activated sludge process. The combined purification process of MBR and ozonation produced an additional COD decrease of 10-15%, and made the Total Surfactants values to comply to the specific legislation. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Exploiting interfacial water properties for desalination and purification applications.

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Xu, Hongwu; Varma, Sameer; Nyman, May Devan

    2008-09-01

    A molecular-scale interpretation of interfacial processes is often downplayed in the analysis of traditional water treatment methods. However, such an approach is critical for the development of enhanced performance in traditional desalination and water treatments. Water confined between surfaces, within channels, or in pores is ubiquitous in technology and nature. Its physical and chemical properties in such environments are unpredictably different from bulk water. As a result, advances in water desalination and purification methods may be accomplished through an improved analysis of water behavior in these challenging environments using state-of-the-art microscopy, spectroscopy, experimental, and computational methods.

  7. Non-Equilibrium Plasma Applications for Water Purification Supporting Human Spaceflight and Terrestrial Point-of-Use

    NASA Technical Reports Server (NTRS)

    Blankson, Isaiah M.; Foster, John E.; Adamovsky, Grigory

    2016-01-01

    2016 NASA Glenn Technology Day Panel Presentation on May 24, 2016. The panel description is: Environmental Impact: NASA Glenn Water Capabilities Both global water scarcity and water treatment concerns are two of the most predominant environmental issues of our time. Glenn researchers share insights on a snow sensing technique, hyper spectral imaging of Lake Erie algal blooms, and a discussion on non-equilibrium plasma applications for water purification supporting human spaceflight and terrestrial point-of-use. The panel moderator will be Bryan Stubbs, Executive Director of the Cleveland Water Alliance.

  8. Cleaner Landfills

    NASA Technical Reports Server (NTRS)

    2000-01-01

    Osmotek, Inc. developed the Direct Osmosis treatment system through SBIR funding from Ames Research Center. Using technology originally developed for flight aboard the Space Station, the company brought it to their commercial water purification treatment system, Direct Osmosis. This water purification system uses a direct osmosis process followed by a reverse osmosis treatment. Because the product extracts water from a waste product, Osmotek is marketing the unit for use in landfills. The system can treat leachate (toxic chemicals leached into a water source), by filtering the water and leaving behind the leahcate. The leachate then becomes solidified into substance that can not seep into water.

  9. Hormone purification by isoelectric focusing in space

    NASA Technical Reports Server (NTRS)

    Bier, M.

    1982-01-01

    The performance of a ground-prototype of an apparatus for recycling isoelectric focusing was evaluated in an effort to provide technology for large scale purification of peptide hormones, proteins, and other biologicals. Special emphasis was given to the effects of gravity on the function of the apparatus and to the determination of potential advantages deriveable from its use in a microgravity environment. A theoretical model of isoelectric focusing sing chemically defined buffer systems for the establishment of the pH gradients was developed. The model was transformed to a form suitable for computer simulations and was used extensively for the design of experimental buffers.

  10. Nanomaterials and Water Purification: Opportunities and Challenges

    NASA Astrophysics Data System (ADS)

    Savage, Nora; Diallo, Mamadou S.

    2005-10-01

    Advances in nanoscale science and engineering suggest that many of the current problems involving water quality could be resolved or greatly ameliorated using nanosorbents, nanocatalysts, bioactive nanoparticles, nanostructured catalytic membranes and nanoparticle enhanced filtration among other products and processes resulting from the development of nanotechnology. Innovations in the development of novel technologies to desalinate water are among the most exciting and promising. Additionally, nanotechnology-derived products that reduce the concentrations of toxic compounds to sub-ppb levels can assist in the attainment of water quality standards and health advisories. This article gives an overview of the use of nanomaterials in water purification. We highlight recent advances on the development of novel nanoscale materials and processes for treatment of surface water, groundwater and industrial wastewater contaminated by toxic metal ions, radionuclides, organic and inorganic solutes, bacteria and viruses. In addition, we discuss some challenges associated with the development of cost effective and environmentally acceptable functional nanomaterials for water purification.

  11. Rare cancer cell analyzer for whole blood applications: automated nucleic acid purification in a microfluidic disposable card.

    PubMed

    Kokoris, M; Nabavi, M; Lancaster, C; Clemmens, J; Maloney, P; Capadanno, J; Gerdes, J; Battrell, C F

    2005-09-01

    One current challenge facing point-of-care cancer detection is that existing methods make it difficult, time consuming and too costly to (1) collect relevant cell types directly from a patient sample, such as blood and (2) rapidly assay those cell types to determine the presence or absence of a particular type of cancer. We present a proof of principle method for an integrated, sample-to-result, point-of-care detection device that employs microfluidics technology, accepted assays, and a silica membrane for total RNA purification on a disposable, credit card sized laboratory-on-card ('lab card") device in which results are obtained in minutes. Both yield and quality of on-card purified total RNA, as determined by both LightCycler and standard reverse transcriptase amplification of G6PDH and BCR-ABL transcripts, were found to be better than or equal to accepted standard purification methods.

  12. Single pass tangential flow filtration to debottleneck downstream processing for therapeutic antibody production.

    PubMed

    Dizon-Maspat, Jemelle; Bourret, Justin; D'Agostini, Anna; Li, Feng

    2012-04-01

    As the therapeutic monoclonal antibody (mAb) market continues to grow, optimizing production processes is becoming more critical in improving efficiencies and reducing cost-of-goods in large-scale production. With the recent trends of increasing cell culture titers from upstream process improvements, downstream capacity has become the bottleneck in many existing manufacturing facilities. Single Pass Tangential Flow Filtration (SPTFF) is an emerging technology, which is potentially useful in debottlenecking downstream capacity, especially when the pool tank size is a limiting factor. It can be integrated as part of an existing purification process, after a column chromatography step or a filtration step, without introducing a new unit operation. In this study, SPTFF technology was systematically evaluated for reducing process intermediate volumes from 2× to 10× with multiple mAbs and the impact of SPTFF on product quality, and process yield was analyzed. Finally, the potential fit into the typical 3-column industry platform antibody purification process and its implementation in a commercial scale manufacturing facility were also evaluated. Our data indicate that using SPTFF to concentrate protein pools is a simple, flexible, and robust operation, which can be implemented at various scales to improve antibody purification process capacity. Copyright © 2011 Wiley Periodicals, Inc.

  13. Membrane-Based Technologies in the Pharmaceutical Industry and Continuous Production of Polymer-Coated Crystals/Particles.

    PubMed

    Chen, Dengyue; Sirkar, Kamalesh K; Jin, Chi; Singh, Dhananjay; Pfeffer, Robert

    2017-01-01

    Membrane technologies are of increasing importance in a variety of separation and purification applications involving liquid phases and gaseous mixtures. Although the most widely used applications at this time are in water treatment including desalination, there are many applications in chemical, food, healthcare, paper and petrochemical industries. This brief review is concerned with existing and emerging applications of various membrane technologies in the pharmaceutical and biopharmaceutical industry. The goal of this review article is to identify important membrane processes and techniques which are being used or proposed to be used in the pharmaceutical and biopharmaceutical operations. How novel membrane processes can be useful for delivery of crystalline/particulate drugs is also of interest. Membrane separation technologies are extensively used in downstream processes for bio-pharmaceutical separation and purification operations via microfiltration, ultrafiltration and diafiltration. Also the new technique of membrane chromatography allows efficient purification of monoclonal antibodies. Membrane filtration techniques of reverse osmosis and nanofiltration are being combined with bioreactors and advanced oxidation processes to treat wastewaters from pharmaceutical plants. Nanofiltration with organic solvent-stable membranes can implement solvent exchange and catalyst recovery during organic solvent-based drug synthesis of pharmaceutical compounds/intermediates. Membranes in the form of hollow fibers can be conveniently used to implement crystallization of pharmaceutical compounds. The novel crystallization methods of solid hollow fiber cooling crystallizer (SHFCC) and porous hollow fiber anti-solvent crystallization (PHFAC) are being developed to provide efficient methods for continuous production of polymer-coated drug crystals in the area of drug delivery. This brief review provides a general introduction to various applications of membrane technologies in the pharmaceutical/biopharmaceutical industry with special emphasis on novel membrane techniques for pharmaceutical applications. The method of coating a drug particle with a polymer using the SHFCC method is stable and ready for scale-up for operation over an extended period. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  14. Preparative isolation and purification of astaxanthin from the microalga Chlorococcum sp. by high-speed counter-current chromatography.

    PubMed

    Li, H B; Chen, F

    2001-08-03

    High-speed counter-current chromatography was applied to the isolation and purification of astaxanthin from microalgae. The crude astaxanthin was obtained by extraction with organic solvents after the astaxanthin esters were saponified. Preparative high-speed counter-current chromatography with a two-phase solvent system composed of n-hexane-ethyl acetate-ethanol-water (5:5:6.5:3, v/v) was successfully performed yielding astaxanthin at 97% purity from 250 mg of the crude extract in a one-step separation.

  15. Protein body-inducing fusions for high-level production and purification of recombinant proteins in plants.

    PubMed

    Conley, Andrew J; Joensuu, Jussi J; Richman, Alex; Menassa, Rima

    2011-05-01

    For the past two decades, therapeutic and industrially important proteins have been expressed in plants with varying levels of success. The two major challenges hindering the economical production of plant-made recombinant proteins include inadequate accumulation levels and the lack of efficient purification methods. To address these limitations, several fusion protein strategies have been recently developed to significantly enhance the production yield of plant-made recombinant proteins, while simultaneously assisting in their subsequent purification. Elastin-like polypeptides are thermally responsive biopolymers composed of a repeating pentapeptide 'VPGXG' sequence that are valuable for the purification of recombinant proteins. Hydrophobins are small fungal proteins capable of altering the hydrophobicity of their respective fusion partner, thus enabling efficient purification by surfactant-based aqueous two-phase systems. Zera, a domain of the maize seed storage protein γ-zein, can induce the formation of protein storage bodies, thus facilitating the recovery of fused proteins using density-based separation methods. These three novel protein fusion systems have also been shown to enhance the accumulation of a range of different recombinant proteins, while concurrently inducing the formation of protein bodies. The packing of these fusion proteins into protein bodies may exclude the recombinant protein from normal physiological turnover. Furthermore, these systems allow for quick, simple and inexpensive nonchromatographic purification of the recombinant protein, which can be scaled up to industrial levels of protein production. This review will focus on the similarities and differences of these artificial storage organelles, their biogenesis and their implication for the production of recombinant proteins in plants and their subsequent purification. © 2011 The Authors. Plant Biotechnology Journal © 2011 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd.

  16. [Study on extraction and purification technology of Hubei ophiopogon saponins].

    PubMed

    Lin, Yun-Han; Li, Chong-Ming; Li, Xiao-Dong; Xiang, Yang; Zhang, Ya-Qin; Zhang, Xiao-Cun; Liu, Xia

    2013-05-01

    To explore the extraction and purification technology of total saponins from the effective parts of Liriope spicata. Orthogonal design was used. Macroporous resin was selected to separate and purify total saponin from the effective parts of Liriope spicata. The process validation was conducted. The total saponins was determined by Ultraviolet Spectrophotometry. The optimal extraction conditions were as follows: 10 times the amount of ethanol (70%) for each occasion and hot reflux (3 x 2 h). Total saponins was purified by D101 macroporous resin. The concentration of eluting ethanol was 50% - 70%. Ethanol (70%) was selected as the best eluent. The result of process validation was consistent with the study. The process is simple and stable enough to significantly improve the extraction rate of the effective parts. The study can provide reference for the research and production of effective parts of traditional Chinese medicine such as Liriope spicata.

  17. Affinity purification of bacterial outer membrane vesicles (OMVs) utilizing a His-tag mutant.

    PubMed

    Alves, Nathan J; Turner, Kendrick B; DiVito, Kyle A; Daniele, Michael A; Walper, Scott A

    To facilitate the rapid purification of bacterial outer membrane vesicles (OMVs), we developed two plasmid constructs that utilize a truncated, transmembrane protein to present an exterior histidine repeat sequence. We chose OmpA, a highly abundant porin protein, as the protein scaffold and utilized the lac promoter to allow for inducible control of the epitope-presenting construct. OMVs containing mutant OmpA-His6 were purified directly from Escherichia coli culture media on an immobilized metal affinity chromatography (IMAC) Ni-NTA resin. This enabling technology can be combined with other molecular tools directed at OMV packaging to facilitate the separation of modified/cargo-loaded OMV from their wt counterparts. In addition to numerous applications in the pharmaceutical and environmental remediation industries, this technology can be utilized to enhance basic research capabilities in the area of elucidating endogenous OMV function. Published by Elsevier Masson SAS.

  18. Calculation of Purification Systems of Hydrocarbonmoderated Reactors; CALCULO DE SISTEMAS DE PURIFICATION DE REACTORES MODERADOS FOR HIDROCARBUROS

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Santos, A.A.

    1958-01-01

    culation of Purification Systems of Hydrocarbonmoderated Reactors). Agustin Alonso Santos. 1958. 23p. As as introduction to the calculation of the purification systems of bydrocarbon-moderated reactors, the effects of heat and radiation on the polyphenols are considered. The chemical, physical, and nuclear properties are tabulated. The formation velocity of the polymers and gases, pyrolysis, effects of heat on the polymer, and the activity accumulated in the moderator ars discussed. The calculation is based on the hypetheses that the radiation catalyzes the formation of polymers, the velocity of the polymerization reaction is constant, the polymer concentration is maintained at a limit whichmore » does not adversely affect the heat transfer properties, the velocity of the separation of polymers in the distillation column is in proportion to their concentration in the hydrocarbon and the pyrolysis causes gaseous products. Formulas are derived expressing the purified flow and the activities accumulated in the distillation residues. The results are applied to the parification system of the Organic Moderated Reactor Experiment (J.S.R.)« less

  19. Low temperature engineering applied to lunar in-situ resource utilization

    NASA Technical Reports Server (NTRS)

    Zhang, Burt; Chui, Talso; Croonquist, Arvid

    2005-01-01

    In support of NASA's Exploration Mission low-temperature scientists and engineers have investigated the process of extracting volatile materials from the lunar regolith, their purification/liquefaction, and storage.

  20. Antifouling membranes for sustainable water purification: strategies and mechanisms.

    PubMed

    Zhang, Runnan; Liu, Yanan; He, Mingrui; Su, Yanlei; Zhao, Xueting; Elimelech, Menachem; Jiang, Zhongyi

    2016-10-24

    One of the greatest challenges to the sustainability of modern society is an inadequate supply of clean water. Due to its energy-saving and cost-effective features, membrane technology has become an indispensable platform technology for water purification, including seawater and brackish water desalination as well as municipal or industrial wastewater treatment. However, membrane fouling, which arises from the nonspecific interaction between membrane surface and foulants, significantly impedes the efficient application of membrane technology. Preparing antifouling membranes is a fundamental strategy to deal with pervasive fouling problems from a variety of foulants. In recent years, major advancements have been made in membrane preparation techniques and in elucidating the antifouling mechanisms of membrane processes, including ultrafiltration, nanofiltration, reverse osmosis and forward osmosis. This review will first introduce the major foulants and the principal mechanisms of membrane fouling, and then highlight the development, current status and future prospects of antifouling membranes, including antifouling strategies, preparation techniques and practical applications. In particular, the strategies and mechanisms for antifouling membranes, including passive fouling resistance and fouling release, active off-surface and on-surface strategies, will be proposed and discussed extensively.

  1. 77 FR 12082 - Notice of Determinations Regarding Eligibility To Apply for Worker Adjustment Assistance

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-28

    ..., Including Leased Workers: Keystone Staffing, Aerotek Staffing. 81,296 Pentair Water Filtration Monticello, IN February 3, 2011. Indiana, LLC, Water Purification Division, Manpower. The following...

  2. High-throughput Cloning and Expression of Integral Membrane Proteins in Escherichia coli

    PubMed Central

    Bruni, Renato

    2014-01-01

    Recently, several structural genomics centers have been established and a remarkable number of three-dimensional structures of soluble proteins have been solved. For membrane proteins, the number of structures solved has been significantly trailing those for their soluble counterparts, not least because over-expression and purification of membrane proteins is a much more arduous process. By using high throughput technologies, a large number of membrane protein targets can be screened simultaneously and a greater number of expression and purification conditions can be employed, leading to a higher probability of successfully determining the structure of membrane proteins. This unit describes the cloning, expression and screening of membrane proteins using high throughput methodologies developed in our laboratory. Basic Protocol 1 deals with the cloning of inserts into expression vectors by ligation-independent cloning. Basic Protocol 2 describes the expression and purification of the target proteins on a miniscale. Lastly, for the targets that express at the miniscale, basic protocols 3 and 4 outline the methods employed for the expression and purification of targets at the midi-scale, as well as a procedure for detergent screening and identification of detergent(s) in which the target protein is stable. PMID:24510647

  3. The modified swirl sedimentation tanks for water purification.

    PubMed

    Ochowiak, Marek; Matuszak, Magdalena; Włodarczak, Sylwia; Ancukiewicz, Małgorzata; Krupińska, Andżelika

    2017-03-15

    This paper discusses design, evaluation, and application for the use of swirl/vortex technologies as liquid purification system. A study was performed using modified swirl sedimentation tanks. The vortex separators (OW, OWK, OWR and OWKR) have been studied under laboratory conditions at liquid flow rate from 2.8⋅10 -5 to 5.1⋅10 -4 [m 3 /s]. The pressure drop and the efficiency of purification of liquid stream were analyzed. The suspended particles of different diameters were successfully removed from liquid with the application of swirl chambers of proposed constructions. It was found that damming of liquid in the tank increases alongside liquid stream at the inlet and depends on the tank construction. The efficiency of the sedimentation tanks increases alongside the diameters of solid particles and decrease in the liquid flow rate. The best construction proved to be the OWR sedimentation tank due to smallest liquid damming, even at high flow rates, and the highest efficiency of the purification liquid stream for solid particles of the smallest diameter. The proposed solution is an alternative to the classical constructions of sedimentation tanks. Copyright © 2016 Elsevier Ltd. All rights reserved.

  4. 75 FR 56059 - Patent Examiner Technical Training Program

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-15

    ...); statistical methods in validation of microarry data; personalized medicine, manufacture of carbon nanospheres... processing, growing monocrystals, hydrogen production, liquid and gas purification and separation, making... Systems and Components: Mixed signal design and architecture, flexible displays, OLED display technology...

  5. Improving recombinant protein purification yield

    USDA-ARS?s Scientific Manuscript database

    Production of adequate amounts of recombinant proteins is essential for antibody production, biochemical activity study, and structural determination during the post-genomic era. It’s technologically challenging and a limiting factor for tung oil research because analytical reagents such as high qua...

  6. Point-of-care technologies for molecular diagnostics using a drop of blood.

    PubMed

    Song, Yujun; Huang, Yu-Yen; Liu, Xuewu; Zhang, Xiaojing; Ferrari, Mauro; Qin, Lidong

    2014-03-01

    Molecular diagnostics is crucial for prevention, identification, and treatment of disease. Traditional technologies for molecular diagnostics using blood are limited to laboratory use because they rely on sample purification and sophisticated instruments, are labor and time intensive, expensive, and require highly trained operators. This review discusses the frontiers of point-of-care (POC) diagnostic technologies using a drop of blood obtained from a finger prick. These technologies, including emerging biotechnologies, nanotechnologies, and microfluidics, hold the potential for rapid, accurate, and inexpensive disease diagnostics. Copyright © 2014 Elsevier Ltd. All rights reserved.

  7. Residual bovine serum albumin (BSA) quantitation in vaccines using automated Capillary Western technology.

    PubMed

    Loughney, John W; Lancaster, Catherine; Ha, Sha; Rustandi, Richard R

    2014-09-15

    Bovine serum albumin (BSA) is a major component of fetal bovine serum (FBS), which is commonly used as a culture medium during vaccine production. Because BSA can cause allergic reactions in humans the World Health Organization (WHO) has set a guidance of 50 ng or less residual BSA per vaccine dose. Vaccine manufacturers are expected to develop sensitive assays to detect residual BSA. Generally, sandwich enzyme-linked immunosorbent assays (ELISA) are used in the industry to detect these low levels of BSA. We report the development of a new improved method for residual BSA detection using the SimpleWestern technology to analyze residual BSA in an attenuated virus vaccine. The method is based on automated Capillary Western and has linearity of two logs, >80% spike recovery (accuracy), intermediate precision of CV <15%, and LOQ of 5.2 ng/ml. The final method was applied to analyze BSA in four lots of bulk vaccine products and was used to monitor BSA clearance during vaccine process purification. Copyright © 2014 Elsevier Inc. All rights reserved.

  8. The Joe 2010 Joint Operating Environment

    DTIC Science & Technology

    2010-02-18

    mitigate the effects of water pollution. By the year 2030, advances in technology may provide many nations with more efficient, sustainable, and...both the number of forces and the time required to rebuild or improve water infrastructure, leave-behind water -purification technology could contribute...represent two of the most important aspects of military effectiveness . The former occurs during peace, when there is time available to think through

  9. Prospective technologies for using biohydrogen in power installations on the basis of fuel cells (a review)

    NASA Astrophysics Data System (ADS)

    Dunikov, D. O.; Borzenko, V. I.; Malyshenko, S. P.; Blinov, D. V.; Kazakov, A. N.

    2013-03-01

    The present state of technology for obtaining hydrogen by biological methods and for purifying it is reviewed from the viewpoint of its possible use in kilowatt-class power installations. Hybrid membranesorption biohydrogen purification methods combining membrane-based pretreatment and sorption-based final treatment, also with the use of metal hydrides, should be regarded as the most efficient ones.

  10. Review of desulfurization process for biogas purification

    NASA Astrophysics Data System (ADS)

    Xiao, Cong; Ma, Yunqian; Ji, Dandan; Zang, Lihua

    2017-12-01

    Hydrogen sulfide (H2S) is a toxic and odorous compound present in biogas produced by the anaerobic digestion of biosolids and other organic materials. Elimination of H2S is necessary as it is extremely hazardous to human health, poisonous to process catalysts and corrosive to equipment. The desulfurization technology is an important part for efficient utilization of biogas. In this paper, the traditional wet and dry desulfurization technology for biogas was reviewed, and the new research progress of biological desulfurization technologies are also introduced.

  11. Application of reverse osmosis in purifying drinking water

    NASA Astrophysics Data System (ADS)

    Jiang, Lei; Tu, Yue; Li, Xiangmin; Li, Haixiang

    2018-06-01

    In view of the shortage of water resources and the pollution of water environment, the development of efficient water purification technology is one of the effective ways to ensure the safety of drinking water. The technology of reverse osmosis has attracted much attention in the application of drinking water security for its advantages of no phase change and simple equipment. This paper briefly introduces the research progress at home and abroad, basic principles, process flow diagram, advantages and disadvantages and development direction of the technology.

  12. Recent progress in the applications of layer-by-layer assembly to the preparation of nanostructured ion-rejecting water purification membranes.

    PubMed

    Sanyal, Oishi; Lee, Ilsoon

    2014-03-01

    Reverse osmosis (RO) and nanofiltration (NF) are the two dominant membrane separation processes responsible for ion rejection. While RO is highly efficient in removal of ions it needs a high operating pressure and offers very low selectivity between ions. Nanofiltration on the other hand has a comparatively low operating pressure and most commercial membranes offer selectivity in terms of ion rejection. However in many nanofiltration operations rejection of monovalent ions is not appreciable. Therefore a high flux high rejection membrane is needed that can be applied to water purification systems. One such alternative is the usage of polyelectrolyte multilayer membranes that are prepared by the deposition of alternately charged polyelectrolytes via layer-by-layer (LbL) assembly method. LbL is one of the most common self-assembly techniques and finds application in various areas. It has a number of tunable parameters like deposition conditions, number of bilayers deposited etc. which can be manipulated as per the type of application. This technique can be applied to make a nanothin membrane skin which gives high rejection and at the same time allow a high water flux across it. Several research groups have applied this highly versatile technique to prepare membranes that can be employed for water purification. Some of these membranes have shown better performance than the commercial nanofiltration and reverse osmosis membranes. These membranes have the potential to be applied to various different aspects of water treatment like water softening, desalination and recovery of certain ions. Besides the conventional method of LbL technique other alternative methods have also been suggested that can make the technique fast, more efficient and thereby make it more commercially acceptable.

  13. Partial Purification of a Legume Nodulation Factor Present in Coconut Water 1

    PubMed Central

    Schaffer, A. G.; Alexander, M.

    1967-01-01

    The nodulation of adventitious roots growing from segments of bean hypocotyl tissue was used as a bioassay for the material present in coconut water which stimulated nodulation. The active material in coconut water is acidic, but it was not possible to extract it from an acid solution with organic solvents. A purification of approximately 70-fold (on a dry wt basis) was obtained using activated charcoal, but at least 10 different compounds were present in the active fractions. A purified fraction of coconut water, which is stimulatory to the growth of carrot root explants, was active in the nodulation assay at a concentration of 2 μg/ml. This represents a 4000-fold purification of the diffusible fraction of coconut water. The charcoal fractionation procedure can be applied to the active material present in extracts of bean leaves. PMID:16656538

  14. Method and apparatus for capacitive deionization and electrochemical purification and regeneration of electrodes

    DOEpatents

    Tran, Tri D.; Farmer, Joseph C.; Murguia, Laura

    2001-01-01

    An electrically regeneratable electrochemical cell (30) for capacitive deionization and electrochemical purification and regeneration of electrodes includes two end plates (31, 32), one at each end of the cell (30). A new regeneration method is applied to the cell (30) which includes slowing or stopping the purification cycle, electrically desorbing contaminants and removing the desorbed contaminants. The cell (30) further includes a plurality of generally identical double-sided intermediate electrodes (37-43) that are equidistally separated from each other, between the two end electrodes (35, 36). As the electrolyte enters the cell, it flows through a continuous open serpentine channel (65-71) defined by the electrodes, substantially parallel to the surfaces of the electrodes. By polarizing the cell (30), ions are removed from the electrolyte and are held in the electric double layers formed at the carbon aerogel surfaces of the electrodes. The cell (30) is regenerated electrically to desorb such previously removed ions.

  15. Soft-Bake Purification of SWCNTs Produced by Pulsed Laser Vaporization

    NASA Technical Reports Server (NTRS)

    Yowell, Leonard; Nikolaev, Pavel; Gorelik, Olga; Allada, Rama Kumar; Sosa, Edward; Arepalli, Sivaram

    2013-01-01

    The "soft-bake" method is a simple and reliable initial purification step first proposed by researchers at Rice University for single-walled carbon nanotubes (SWCNT) produced by high-pressure carbon mon oxide disproportionation (HiPco). Soft-baking consists of annealing as-produced (raw) SWCNT, at low temperatures in humid air, in order to degrade the heavy graphitic shells that surround metal particle impurities. Once these shells are cracked open by the expansion and slow oxidation of the metal particles, the metal impurities can be digested through treatment with hydrochloric acid. The soft-baking of SWCNT produced by pulsed-laser vaporization (PLV) is not straightforward, because the larger average SWCNT diameters (.1.4 nm) and heavier graphitic shells surrounding metal particles call for increased temperatures during soft-bake. A part of the technology development focused on optimizing the temperature so that effective cracking of the graphitic shells is balanced with maintaining a reasonable yield, which was a critical aspect of this study. Once the ideal temperature was determined, a number of samples of raw SWCNT were purified using the soft-bake method. An important benefit to this process is the reduced time and effort required for soft-bake versus the standard purification route for SWCNT. The total time spent purifying samples by soft-bake is one week per batch, which equates to a factor of three reduction in the time required for purification as compared to the standard acid purification method. Reduction of the number of steps also appears to be an important factor in improving reproducibility of yield and purity of SWCNT, as small deviations are likely to get amplified over the course of a complicated multi-step purification process.

  16. Novel Membranes and Systems for Industrial and Municipal Water Purification and Reuse

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    None

    This factsheet describes a project that developed nano-engineered, high-permeance membrane materials with more than double the permeance of current reverse osmosis membranes as well as manufacturing technologies for large-scale production of the novel materials.

  17. Biopharmaceuticals from microorganisms: from production to purification.

    PubMed

    Jozala, Angela Faustino; Geraldes, Danilo Costa; Tundisi, Louise Lacalendola; Feitosa, Valker de Araújo; Breyer, Carlos Alexandre; Cardoso, Samuel Leite; Mazzola, Priscila Gava; Oliveira-Nascimento, Laura de; Rangel-Yagui, Carlota de Oliveira; Magalhães, Pérola de Oliveira; Oliveira, Marcos Antonio de; Pessoa, Adalberto

    2016-12-01

    The use of biopharmaceuticals dates from the 19th century and within 5-10 years, up to 50% of all drugs in development will be biopharmaceuticals. In the 1980s, the biopharmaceutical industry experienced a significant growth in the production and approval of recombinant proteins such as interferons (IFN α, β, and γ) and growth hormones. The production of biopharmaceuticals, known as bioprocess, involves a wide range of techniques. In this review, we discuss the technology involved in the bioprocess and describe the available strategies and main advances in microbial fermentation and purification process to obtain biopharmaceuticals. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  18. Affinity Purification of Tumor Necrosis Factor-α Expressed in Raji Cells by Produced scFv Antibody Coupled CNBr-Activated Sepharose

    PubMed Central

    Abdolalizadeh, Jalal; Majidi Zolbanin, Jafar; Nouri, Mohammad; Baradaran, Behzad; Movassaghpour, AliAkbar; Farajnia, Safar; Omidi, Yadollah

    2013-01-01

    Purpose: Recombinant tumor necrosis factor-alpha (TNF-α) has been utilized as an antineoplastic agent for the treatment of patients with melanoma and sarcoma. It targets tumor cell antigens by impressing tumor-associated vessels. Protein purification with affinity chromatography has been widely used in the downstream processing of pharmaceutical-grade proteins. Methods:In this study, we examined the potential of our produced anti-TNF-α scFv fragments for purification of TNF-α produced by Raji cells. The Raji cells were induced by lipopolysaccharides (LPS) to express TNF-α. Western blotting and Fluorescence-activated cell sorting (FACS) flow cytometry analyses were used to evaluate the TNF-α expression. The anti-TNF-α scFv selected from antibody phage display library was coupled to CNBr-activated sepharose 4B beads used for affinity purification of expressed TNF-α and the purity of the protein was assessed by SDS-PAGE. Results: Western blot and FACS flow cytometry analyses showed the successful expression of TNF-α with Raji cells. SDS-PAGE analysis showed the performance of scFv for purification of TNF-α protein with purity over 95%. Conclusion: These findings confirm not only the potential of the produced scFv antibody fragments but also this highly pure recombinant TNF-α protein can be applied for various in vitro and in vivo applications. PMID:24312807

  19. DOE Office of Scientific and Technical Information (OSTI.GOV)

    G.I. Alekseev

    Wastewater generation, water consumption, and water management at coke plants are considered. Measures to create runoff-free water-supply and sewer systems are discussed. Filters for water purification, corrosion inhibitors, and biocides are described. An integrated single-phase technology for the removal of phenols, thiocyanides, and ammoniacal nitrogen is outlined.

  20. Nanostructured Titanium Oxide Film- And Membrane-Based Photocatalysis For Water Treatment

    EPA Science Inventory

    Titanium Oxide (TiO2) photocatalysis, one of the ultraviolet (UV)-based advanced oxidation technologies (AOTs) and nanotechnologies (AONs), has attracted great attention for the development of efficient water treatment and purification systems due to the effectiveness ...

  1. Using an FPLC to promote active learning of the principles of protein structure and purification.

    PubMed

    Robinson, Rebekah L; Neely, Amy E; Mojadedi, Wais; Threatt, Katie N; Davis, Nicole Y; Weiland, Mitch H

    2017-01-02

    The concepts of protein purification are often taught in undergraduate biology and biochemistry lectures and reinforced during laboratory exercises; however, very few reported activities allow students to directly gain experience using modern protein purification instruments, such as Fast Protein Liquid Chromatography (FPLC). This laboratory exercise uses size exclusion chromatography (SEC) and ion exchange (IEX) chromatography to separate a mixture of four different proteins. Students use an SEC chromatogram and corresponding SDS-PAGE gel to understand how protein conformations change under different conditions (i.e. native and non-native). Students explore strategies to separate co-eluting proteins by IEX chromatography. Using either cation or anion exchange, one protein is bound to the column while the other is collected in the flow-through. In this exercise, undergraduate students gain hands-on experience with experimental design, buffer and sample preparation, and implementation of instrumentation that is commonly used by experienced researchers while learning and applying the fundamental concepts of protein structure, protein purification, and SDS-PAGE. © 2016 by The International Union of Biochemistry and Molecular Biology, 45(1):60-68, 2017. © 2016 The International Union of Biochemistry and Molecular Biology.

  2. Purification processes of xenogeneic bone substitutes and their impact on tissue reactions and regeneration.

    PubMed

    Perić Kačarević, Zeljka; Kavehei, Faraz; Houshmand, Alireza; Franke, Jörg; Smeets, Ralf; Rimashevskiy, Denis; Wenisch, Sabine; Schnettler, Reinhard; Jung, Ole; Barbeck, Mike

    2018-04-01

    Xenogeneic bone substitute materials are widely used in oral implantology. Prior to their clinical use, purification of the former bone tissue has to be conducted to ensure the removal of immunogenic components and pathogens. Different physicochemical methods are applied for purification of the donor tissue, and temperature treatment is one of these methods. Differences in these methods and especially the application of different temperatures for purification may lead to different material characteristics, which may influence the tissue reactions to these materials and the related (bone) healing process. However, little is known about the different material characteristics and their influences on the healing process. Thus, the aim of this mini-review is to summarize the preparation processes and the related material characteristics, safety aspects, tissue reactions, resorbability and preclinical and clinical data of two widely used xenogeneic bone substitutes that mainly differ in the temperature treatment: sintered (cerabone ® ) and non-sintered (Bio-Oss ® ) bovine-bone materials. Based on the summarized data from the literature, a connection between the material-induced tissue reactions and the consequences for the healing processes are presented with the aim of translation into their clinical application.

  3. Waste water purification using new porous ceramics prepared by recycling waste glass and bamboo charcoal

    NASA Astrophysics Data System (ADS)

    Nishida, Tetsuaki; Morimoto, Akane; Yamamoto, Yoshito; Kubuki, Shiro

    2017-12-01

    New porous ceramics (PC) prepared by recycling waste glass bottle of soft drinks (80 mass%) and bamboo charcoal (20 mass%) without any binder was applied to the waste water purification under aeration at 25 °C. Artificial waste water (15 L) containing 10 mL of milk was examined by combining 15 mL of activated sludge and 750 g of PC. Biochemical oxygen demand (BOD) showed a marked decrease from 178 to 4.0 (±0.1) mg L-1 in 5 days and to 2.0 (±0.1) mg L-1 in 7 days, which was equal to the Environmental Standard for the river water (class A) in Japan. Similarly, chemical oxygen demand (COD) decreased from 158 to 3.6 (±0.1) mg L-1 in 5 days and to 2.2 (±0.1) mg L-1 in 9 days, which was less than the Environmental Standard for the Seawater (class B) in Japan: 3.0 mg L-1. These results prove the high water purification ability of the PC, which will be effectively utilized for the purification of drinking water, fish preserve water, fish farm water, etc.

  4. Evaluation of immobilized metal-ion affinity chromatography (IMAC) as a technique for IgG(1) monoclonal antibodies purification: the effect of chelating ligand and support.

    PubMed

    Bresolin, I T L; Borsoi-Ribeiro, M; Tamashiro, W M S C; Augusto, E F P; Vijayalakshmi, M A; Bueno, S M A

    2010-04-01

    Monoclonal antibodies (MAbs) have been used for therapies and some analytical procedures as highly purified molecules. Many techniques have been applied and studied, focusing on monoclonal antibodies purification. In this study, an immobilized metal affinity chromatography membrane was developed and evaluated for the purification of anti-TNP IgG(1) mouse MAbs from cell culture supernatant after precipitation with a 50% saturated ammonium sulfate solution. The chelating ligands iminodiacetic acid, carboxymethylated aspartic acid (CM-Asp), nitrilotriacetic acid, and tris (carboxymethyl) ethylenediamine in agarose gels with immobilized Ni(II) and Zn(II) ions were compared for the adsorption and desorption of MAbs. The most promising chelating ligand--CM-Asp--was then coupled to poly(ethylene vinyl alcohol) (PEVA) hollow fiber membranes. According to SDS-PAGE and ELISA analyses, a higher selectivity and a purification factor of 85.9 (fraction eluted at 500 mM Tris) were obtained for IgG(1) using PEVA-CM-Asp-Zn(II). The anti-TNP MAb could be eluted under mild pH conditions causing no loss of antigen binding capacity.

  5. Two novel solvent system compositions for protected synthetic peptide purification by centrifugal partition chromatography.

    PubMed

    Amarouche, Nassima; Giraud, Matthieu; Forni, Luciano; Butte, Alessandro; Edwards, F; Borie, Nicolas; Renault, Jean-Hugues

    2014-04-11

    Protected synthetic peptide intermediates are often hydrophobic and not soluble in most common solvents. They are thus difficult to purify by preparative reversed-phase high-performance liquid chromatography (RP-HPLC), usually used for industrial production. It is then challenging to develop alternative chromatographic purification processes. Support-free liquid-liquid chromatographic techniques, including both hydrostatic (centrifugal partition chromatography or CPC) and hydrodynamic (counter-current chromatography or CCC) devices, are mainly involved in phytochemical studies but have also been applied to synthetic peptide purification. In this framework, two new biphasic solvent system compositions covering a wide range of polarity were developed to overcome solubility problems mentioned above. The new systems composed of heptane/tetrahydrofuran/acetonitrile/dimethylsulfoxide/water and heptane/methyl-tetrahydrofuran/N-methylpyrrolidone/water were efficiently used for the CPC purification of a 39-mer protected exenatide (Byetta®) and a 8-mer protected peptide intermediate of bivalirudin (Angiox®) synthesis. Phase compositions of the different biphasic solvent systems were determined by (1)H nuclear magnetic resonance. Physico-chemical properties including viscosity, density and interfacial tension of these biphasic systems are also described. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Study on the extraction and purification of glycoprotein from the yellow seahorse, Hippocampus kuda Bleeker

    PubMed Central

    Su, Yuting; Xu, Yongjian

    2015-01-01

    The optimum parameters of extraction for glycoprotein from seahorse were examined and determined by Box-Behnken combined with ultrasonic extraction technology. Column chromatography of glycoprotein was used for further purification. The optimal extraction conditions of seahorse glycoprotein were extracting time 4.3 h, salt concentration 0.08 mol/L, extracting temperature 73°C, raw material, and water ratio 1:6. At the optimal conditions, the yield of saccharide reached to 1.123%, and the yield of protein reached to 5.898%. For purifying the crude glycoprotein, the stage renounces of DEAE-52 column chromatography were done, respectively, with 0.05, 0.1, 0.5 mol/L NaHCO3 solution, and further purification was done with Sephadex G-100 column chromatography. Finally, two pieces of seahorse glycoprotein were obtained by the column chromatography, that is, HG-11 and HG-21. The saccharide content was 56.7975% and 39.479%, the protein content was 30.5475% and 51.747%, respectively. PMID:26288722

  7. Protein complex purification from Thermoplasma acidophilum using a phage display library.

    PubMed

    Hubert, Agnes; Mitani, Yasuo; Tamura, Tomohiro; Boicu, Marius; Nagy, István

    2014-03-01

    We developed a novel protein complex isolation method using a single-chain variable fragment (scFv) based phage display library in a two-step purification procedure. We adapted the antibody-based phage display technology which has been developed for single target proteins to a protein mixture containing about 300 proteins, mostly subunits of Thermoplasma acidophilum complexes. T. acidophilum protein specific phages were selected and corresponding scFvs were expressed in Escherichia coli. E. coli cell lysate containing the expressed His-tagged scFv specific against one antigen protein and T. acidophilum crude cell lysate containing intact target protein complexes were mixed, incubated and subjected to protein purification using affinity and size exclusion chromatography steps. This method was confirmed to isolate intact particles of thermosome and proteasome suitable for electron microscopy analysis and provides a novel protein complex isolation strategy applicable to organisms where no genetic tools are available. Copyright © 2013 Elsevier B.V. All rights reserved.

  8. A simple and low-cost platform technology for producing pexiganan antimicrobial peptide in E. coli.

    PubMed

    Zhao, Chun-Xia; Dwyer, Mirjana Dimitrijev; Yu, Alice Lei; Wu, Yang; Fang, Sheng; Middelberg, Anton P J

    2015-05-01

    Antimicrobial peptides, as a new class of antibiotics, have generated tremendous interest as potential alternatives to classical antibiotics. However, the large-scale production of antimicrobial peptides remains a significant challenge. This paper reports a simple and low-cost chromatography-free platform technology for producing antimicrobial peptides in Escherichia coli (E. coli). A fusion protein comprising a variant of the helical biosurfactant protein DAMP4 and the known antimicrobial peptide pexiganan is designed by joining the two polypeptides, at the DNA level, via an acid-sensitive cleavage site. The resulting DAMP4(var)-pexiganan fusion protein expresses at high level and solubility in recombinant E. coli, and a simple heat-purification method was applied to disrupt cells and deliver high-purity DAMP4(var)-pexiganan protein. Simple acid cleavage successfully separated the DAMP4 variant protein and the antimicrobial peptide. Antimicrobial activity tests confirmed that the bio-produced antimicrobial peptide has the same antimicrobial activity as the equivalent product made by conventional chemical peptide synthesis. This simple and low-cost platform technology can be easily adapted to produce other valuable peptide products, and opens a new manufacturing approach for producing antimicrobial peptides at large scale using the tools and approaches of biochemical engineering. © 2014 Wiley Periodicals, Inc.

  9. Effects of influent C/N ratios and treatment technologies on integral biogas upgrading and pollutants removal from synthetic domestic sewage.

    PubMed

    Xu, Jie; Wang, Xue; Sun, Shiqing; Zhao, Yongjun; Hu, Changwei

    2017-09-07

    Three different treatment technologies, namely mono-algae culture, algal-bacterial culture, and algal-fungal culture, were applied to remove pollutants form synthetic domestic sewage and to remove CO 2 from biogas in a photobioreactor. The effects of different initial influent C/N ratios on microalgal growth rates and pollutants removal efficiencies by the three microalgal cultures were investigated. The best biogas upgrading and synthetic domestic sewage pollutants removal effect was achieved in the algal-fungal system at the influent C/N ratio of 5:1. At the influent C/N ratio of 5:1, the algal-fungal system achieved the highest mean chemical oxygen demand (COD) removal efficiency of 81.92% and total phosphorus (TP) removal efficiency of 81.52%, respectively, while the algal-bacterial system demonstrated the highest mean total nitrogen (TN) removal efficiency of 82.28%. The average CH 4 concentration in upgraded biogas and the removal efficiencies of COD, TN, and TP were 93.25 ± 3.84% (v/v), 80.23 ± 3.92%, 75.85 ± 6.61%, and 78.41 ± 3.98%, respectively. These results will provide a reference for wastewater purification ad biogas upgrading with microalgae based technology.

  10. Application of a risk analysis method to different technologies for producing a monoclonal antibody employed in hepatitis B vaccine manufacturing.

    PubMed

    Milá, Lorely; Valdés, Rodolfo; Tamayo, Andrés; Padilla, Sigifredo; Ferro, Williams

    2012-03-01

    CB.Hep-1 monoclonal antibody (mAb) is used for a recombinant Hepatitis B vaccine manufacturing, which is included in a worldwide vaccination program against Hepatitis B disease. The use of this mAb as immunoligand has been addressed into one of the most efficient steps of active pharmaceutical ingredient purification process. Regarding this, Quality Risk Management (QRM) provides an excellent framework for the risk management use in pharmaceutical manufacturing and quality decision-making applications. Consequently, this study sought applying a prospective risk analysis methodology Failure Mode Effects Analysis (FMEA) as QRM tool for analyzing different CB.Hep-1 mAb manufacturing technologies. As main conclusions FMEA was successfully used to assess risks associated with potential problems in CB.Hep-1 mAb manufacturing processes. The severity and occurrence of risks analysis evidenced that the percentage of very high severe risks ranged 31.0-38.7% of all risks and the huge majority of risks have a very low occurrence level (61.9-83.3%) in all assessed technologies. Finally, additive Risk Priority Number, was descending ordered as follow: transgenic plants (2636), ascites (2577), transgenic animals (2046) and hollow fiber bioreactors (1654), which also corroborated that in vitro technology, should be the technology of choice for CB.Hep-1 mAb manufacturing in terms of risks and mAb molecule quality. Copyright © 2011 The International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.

  11. Bio-recognition and functional lipidomics by glycosphingolipid transfer technology

    PubMed Central

    TAKI, Takao

    2013-01-01

    Through glycosphingolipid biochemical research, we developed two types of transcription technologies. One is a biochemical transfer of glycosphingolipids to peptides. The other is a physicochemical transfer of glycosphingolipids in silica gel to the surface of a plastic membrane. Using the first technology, we could prepare peptides which mimic the shapes of glycosphingolipid molecules by biopanning with a phage-displayed peptide library and anti-glycosphingolipid antibodies as templates. The peptides thus obtained showed biological properties and functions similar to those of the original glycosphingolipids, such as lectin binding, glycosidase modulation, inhibition of tumor metastasis and immune response against the original antigen glycosphingolipid, and we named them glyco-replica peptides. The results showed that the newly prepared peptides could be used effectively as a bio-recognition system and suggest that the glyco-replica peptides can be widely applied to therapeutic fields. Using the second technology, we could establish a functional lipidomics with a thin-layer chromatography-blot/matrix-assisted laser desorption ionization-time of flight mass spectrometry (TLC-Blot/MALDI-TOF MS) system. By transferring glycosphingolipids on a plastic membrane surface from a TLC plate, innovative biochemical approaches such as simple purification of individual glycosphingolipids, binding studies, and enzyme reactions could be developed. The combinations of these biochemical approaches and MALDI-TOF MS on the plastic membrane could provide new strategies for glycosphingolipid science and the field of lipidomics. In this review, typical applications of these two transfer technologies are introduced. PMID:23883610

  12. Simple purification method for a recombinantly expressed native His-tag-free aminopeptidase A from Lactobacillus delbrueckii.

    PubMed

    Stressler, Timo; Tanzer, Coralie; Ewert, Jacob; Claaßen, Wolfgang; Fischer, Lutz

    2017-03-01

    The aminopeptidase A (PepA; EC 3.4.11.7) is an intracellular exopeptidase present in lactic acid bacteria. The PepA cleaves glutamyl/aspartyl residues from the N-terminal end of peptides and can, therefore, be applied for the production of protein hydrolysates with an increased amount of these amino acids, which results in a savory taste (umami). The first PepA from a lactobacilli strain was recombinantly expressed in Escherichia coli in a recently published study and harbored a C-terminal His 6 -tag for easier purification. Due to the fact that a His-tag might influence the properties of an enzyme, a simple purification method for the non-His-tagged PepA was required. Surprisingly, the PepA precipitated at a very low ammonium sulfate concentration of 5%. Unusual for a precipitating step, the purity of PepA was over 95% and the obtained activity yield was 110%. The high purity allows biochemical characterization and kinetic investigation. As a result, the optimum pH (6.0-6.5) and temperature (60-65 °C) were comparable to the His 6 -tag harboring PepA; the K M value was at 0.79 mM slightly lower compared to 1.21 mM, respectively. Since PepA is a homo dodecamer, it has a high molecular mass of approximately 480 kDa. Therefore, a subsequent preparative size-exclusion chromatography (SEC) step seemed promising. The PepA after SEC was purified to homogeneity. In summary, the simple two-step purification method presented can be applied to purify high amounts of PepA that will allow the performance of experiments in the future to crystalize PepA for the first time. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. [Studies on the process of Herba Clinopodii saponins purified with macroporous adsorption resin].

    PubMed

    Zhang, Yi; Yan, Dan; Han, Yumei

    2005-10-01

    To study the technological parameters of the purification process of saponins with macroporous adsorption resin. The adsorptive characteristics and elutive parameters of the process were studied by taking the elutive and purified ratio of saponins as markers. 11.4 ml of the extraction of Herba Clinopodii (crude drugs 0.2 g/ml) was purified with a column of macroporous adsorption resin (phi15 mm x H90 mm, dry weight 2.5 g) and washed with 3BV of distilled water, then eluted with 3BV of 30% ethanol and 3BV of 70% ethanol. Most of saponins were collected in the 70% ethanol. With macroporous adsorption resin adsorbing and purifying,the elutive ratio of saponins is 86.8% and the purity reaches 153.2%. So this process of applying macroporous adsorption resin to adsorb and purify Saponins is feasible.

  14. Solar Thermal Energy Storage Device: Hybrid Nanostructures for High-Energy-Density Solar Thermal Fuels

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    None

    2012-01-09

    HEATS Project: MIT is developing a thermal energy storage device that captures energy from the sun; this energy can be stored and released at a later time when it is needed most. Within the device, the absorption of sunlight causes the solar thermal fuel’s photoactive molecules to change shape, which allows energy to be stored within their chemical bonds. A trigger is applied to release the stored energy as heat, where it can be converted into electricity or used directly as heat. The molecules would then revert to their original shape, and can be recharged using sunlight to begin themore » process anew. MIT’s technology would be 100% renewable, rechargeable like a battery, and emissions-free. Devices using these solar thermal fuels—called Hybrisol—can also be used without a grid infrastructure for applications such as de-icing, heating, cooking, and water purification.« less

  15. Redesigning of Microbial Cell Surface and Its Application to Whole-Cell Biocatalysis and Biosensors.

    PubMed

    Han, Lei; Zhao, Yukun; Cui, Shan; Liang, Bo

    2018-06-01

    Microbial cell surface display technology can redesign cell surfaces with functional proteins and peptides to endow cells some unique features. Foreign peptides or proteins are transported out of cells and immobilized on cell surface by fusing with anchoring proteins, which is an effective solution to avoid substance transfer limitation, enzyme purification, and enzyme instability. As the most frequently used prokaryotic and eukaryotic protein surface display system, bacterial and yeast surface display systems have been widely applied in vaccine, biocatalysis, biosensor, bioadsorption, and polypeptide library screening. In this review of bacterial and yeast surface display systems, different cell surface display mechanisms and their applications in biocatalysis as well as biosensors are described with their strengths and shortcomings. In addition to single enzyme display systems, multi-enzyme co-display systems are presented here. Finally, future developments based on our and other previous reports are discussed.

  16. TMV-Gate vectors: Gateway compatible tobacco mosaic virus based expression vectors for functional analysis of proteins

    PubMed Central

    Kagale, Sateesh; Uzuhashi, Shihomi; Wigness, Merek; Bender, Tricia; Yang, Wen; Borhan, M. Hossein; Rozwadowski, Kevin

    2012-01-01

    Plant viral expression vectors are advantageous for high-throughput functional characterization studies of genes due to their capability for rapid, high-level transient expression of proteins. We have constructed a series of tobacco mosaic virus (TMV) based vectors that are compatible with Gateway technology to enable rapid assembly of expression constructs and exploitation of ORFeome collections. In addition to the potential of producing recombinant protein at grams per kilogram FW of leaf tissue, these vectors facilitate either N- or C-terminal fusions to a broad series of epitope tag(s) and fluorescent proteins. We demonstrate the utility of these vectors in affinity purification, immunodetection and subcellular localisation studies. We also apply the vectors to characterize protein-protein interactions and demonstrate their utility in screening plant pathogen effectors. Given its broad utility in defining protein properties, this vector series will serve as a useful resource to expedite gene characterization efforts. PMID:23166857

  17. Optofluidic opportunities in global health, food, water and energy

    NASA Astrophysics Data System (ADS)

    Chen, Yih-Fan; JiangLj, Mm, Aj,; Vo Contributed Equally To This Paper., Li; Mancuso, Matthew; Jain, Aadhar; Oncescu, Vlad; Erickson, David

    2012-07-01

    Optofluidics is a rapidly advancing field that utilizes the integration of optics and microfluidics to provide a number of novel functionalities in microsystems. In this review, we discuss how this approach can potentially be applied to address some of the greatest challenges facing both the developing and developed world, including healthcare, food shortages, malnutrition, water purification, and energy. While medical diagnostics has received most of the attention to date, here we show that some other areas can also potentially benefit from optofluidic technology. Whenever possible we briefly describe how microsystems are currently used to address these problems and then explain why and how optofluidics can provide better solutions. The focus of the article is on the applications of optofluidic techniques in low-resource settings, but we also emphasize that some of these techniques, such as those related to food production, food safety assessment, nutrition monitoring, and energy production, could be very useful in well-developed areas as well.

  18. Collecting, archiving and processing DNA from wildlife samples using FTA® databasing paper

    PubMed Central

    Smith, LM; Burgoyne, LA

    2004-01-01

    Background Methods involving the analysis of nucleic acids have become widespread in the fields of traditional biology and ecology, however the storage and transport of samples collected in the field to the laboratory in such a manner to allow purification of intact nucleic acids can prove problematical. Results FTA® databasing paper is widely used in human forensic analysis for the storage of biological samples and for purification of nucleic acids. The possible uses of FTA® databasing paper in the purification of DNA from samples of wildlife origin were examined, with particular reference to problems expected due to the nature of samples of wildlife origin. The processing of blood and tissue samples, the possibility of excess DNA in blood samples due to nucleated erythrocytes, and the analysis of degraded samples were all examined, as was the question of long term storage of blood samples on FTA® paper. Examples of the end use of the purified DNA are given for all protocols and the rationale behind the processing procedures is also explained to allow the end user to adjust the protocols as required. Conclusions FTA® paper is eminently suitable for collection of, and purification of nucleic acids from, biological samples from a wide range of wildlife species. This technology makes the collection and storage of such samples much simpler. PMID:15072582

  19. [Study on the extraction process and macroporous resin for purification of Timosaponin B II].

    PubMed

    Liu, Yan-Ping; Ding, Yue; Zhang, Tong; Wang, Bing; Cai, Zhen-Zhen; Tao, Jian-Sheng

    2013-06-01

    To optimize the extraction process and macroporous resin for purification of Timosaponin B II from Anemarrhena asphodeloides. Orthogonal design L9 (34) was employed to optimize the circumfluence extraction conditions by taking the extraction yield of Timosaponin B II as index. The absorption-desorption characteristics of eight kinds of macroporous resins were evaluated, then the best resin was chosen to optimize the purification process conditions. The optimum extraction conditions were as follows: the herb was extracted for 2 times (2 hours each time) with 8.5-fold 50% ethanol at the first time and 6-fold 50% ethanol at the second time. HPD100 resin showed a good property for the absorption-desorption of Timosaponin B II. The optimum technological conditions of HPD100 resin were as follows:the solution concentration was 0.23 mg/mL, the amount of saturated adsorption at 4/5 body volumn (BV) resin, the HPD100 resin was washed with 3 BV water and 6 BV 20% ethanol solution to remove the impurity, then the Timosaponin B II was desorbed by 5 BV ethanol solution. The purity of Timosaponin B II was about 50%. The optimized extraction process and purification is stable, efficient and suitable for industrial production.

  20. Inclusion bodies and purification of proteins in biologically active forms.

    PubMed

    Mukhopadhyay, A

    1997-01-01

    Even though recombinant DNA technology has made possible the production of valuable therapeutic proteins, its accumulation in the host cell as inclusion body poses serious problems in the recovery of functionally active proteins. In the last twenty years, alternative techniques have been evolved to purify biologically active proteins from inclusion bodies. Most of these remain only as inventions and very few are commercially exploited. This review summarizes the developments in isolation, refolding and purification of proteins from inclusion bodies that could be used for vaccine and non-vaccine applications. The second section involves a discussion on inclusion bodies, how they are formed, and their physicochemical properties. In vivo protein folding in Escherichia coli and kinetics of in vitro protein folding are the subjects of the third and fourth sections respectively. The next section covers the recovery of bioactive protein from inclusion bodies: it includes isolation of inclusion body from host cell debris, purification in denatured state alternate refolding techniques, and final purification of active molecules. Since purity and safety are two important issues in therapeutic grade proteins, the following three sections are devoted to immunological and biological characterization of biomolecules, nature, and type of impurities normally encountered, and their detection. Lastly, two case studies are discussed to demonstrate the sequence of process steps involved.

  1. [Purification of complicated industrial organic waste gas by complex absorption].

    PubMed

    Chen, Ding-Sheng; Cen, Chao-Ping; Tang, Zhi-Xiong; Fang, Ping; Chen, Zhi-Hang

    2011-12-01

    Complicated industrial organic waste gas with the characteristics of low concentration,high wind volume containing inorganic dust and oil was employed the research object by complex absorption. Complex absorption mechanism, process flow, purification equipment and engineering application were studied. Three different surfactants were prepared for the composite absorbent to purify exhaust gas loaded with toluene and butyl acetate, respectively. Results show that the low surface tension of the composite absorbent can improve the removal efficiency of toluene and butyl acetate. With the advantages of the water film, swirl plate and fill absorption device, efficient absorption equipment was developed for the treatment of complicated industrial organic waste gas. It is with superiorities of simple structure, small size, anti-jam and high mass transfer. Based on absorption technology, waste gas treatment process integrated with heating stripping, burning and anaerobic and other processes, so that emissions of waste gas and absorption solution could meet the discharge standards. The technology has been put into practice, such as manufacturing and spraying enterprises.

  2. Energy-neutral sustainable nutrient recovery incorporated with the wastewater purification process in an enlarged microbial nutrient recovery cell

    NASA Astrophysics Data System (ADS)

    Sun, Dongya; Gao, Yifan; Hou, Dianxun; Zuo, Kuichang; Chen, Xi; Liang, Peng; Zhang, Xiaoyuan; Ren, Zhiyong Jason; Huang, Xia

    2018-04-01

    Recovery of nutrient resources from the wastewater is now an inevitable strategy to maintain the supply of both nutrient and water for our huge population. While the intensive energy consumption in conventional nutrient recovery technologies still remained as the bottleneck towards the sustainable nutrient recycle. This study proposed an enlarged microbial nutrient recovery cell (EMNRC) which was powered by the energy contained in wastewater and achieved multi-cycle nutrient recovery incorporated with in situ wastewater treatment. With the optimal recovery solution of 3 g/L NaCl and the optimal volume ratio of wastewater to recovery solution of 10:1, >89% of phosphorus and >62% of ammonium nitrogen were recovered into struvite. An extremely low water input ratio of <1% was required to obtain the recovered fertilizer and the purified water. It was proved the EMNRC system was a promising technology which could utilize the chemical energy contained in wastewater itself and energy-neutrally recover nutrient during the continuous wastewater purification process.

  3. Water management, purification, and conservation in arid climates. Volume 1: Water management

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Goosen, M.F.A.; Shayya, W.H.

    1999-07-01

    Arid regions are already feeling the severe restraining effects of potable water shortages. In coming years, humid and sub-humid regions of the world will also have to face many of these same problems. In the future, serious conflicts may arise not because of a lack of oil, but due to water shortages. Are there solutions to these problems? Aside from increasing public awareness about the importance of water, society needs to take a three pronged approach: water needs to be effectively managed, it needs to be economically purified, and it needs to be conserved. Only by doing these three thingsmore » in unison can they hope to alleviate the water problems faced by arid regions of the world. This book presents information valuable to seeking, finding and using current technologies to help solve these problems now. Volume 1 examines water management problems in detail, along with water problems and water resources in arid climates, and includes chapters that cover aspects of water management. Water purification technology is another key issue. The economics of this technology is becoming more critical in arid areas due to increasing urbanization and industrialization.« less

  4. Carbon dioxide gas purification and analytical measurement for leading edge 193nm lithography

    NASA Astrophysics Data System (ADS)

    Riddle Vogt, Sarah; Landoni, Cristian; Applegarth, Chuck; Browning, Matt; Succi, Marco; Pirola, Simona; Macchi, Giorgio

    2015-03-01

    The use of purified carbon dioxide (CO2) has become a reality for leading edge 193 nm immersion lithography scanners. Traditionally, both dry and immersion 193 nm lithographic processes have constantly purged the optics stack with ultrahigh purity compressed dry air (UHPCDA). CO2 has been utilized for a similar purpose as UHPCDA. Airborne molecular contamniation (AMC) purification technologies and analytical measurement methods have been extensively developed to support the Lithography Tool Manufacturers purity requirements. This paper covers the analytical tests and characterizations carried out to assess impurity removal from 3.0 N CO2 (beverage grade) for its final utilization in 193 nm and EUV scanners.

  5. Direct Down-scale Experiments of Concentration Column Designs for SHINE Process

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Youker, Amanda J.; Stepinski, Dominique C.; Vandegrift, George F.

    Argonne is assisting SHINE Medical Technologies in their efforts to become a domestic Mo-99 producer. The SHINE accelerator-driven process uses a uranyl-sulfate target solution for the production of fission-product Mo-99. Argonne has developed a molybdenum recovery and purification process for this target solution. The process includes an initial Mo recovery column followed by a concentration column to reduce the product volume from 15-25 L to < 1 L prior to entry into the LEU Modified Cintichem (LMC) process for purification.1 This report discusses direct down-scale experiments of the plant-scale concentration column design, where the effects of loading velocity and temperaturemore » were investigated.« less

  6. Aptamer facilitated purification of functional proteins.

    PubMed

    Beloborodov, Stanislav S; Bao, Jiayin; Krylova, Svetlana M; Shala-Lawrence, Agnesa; Johnson, Philip E; Krylov, Sergey N

    2018-01-15

    DNA aptamers are attractive capture probes for affinity chromatography since, in contrast to antibodies, they can be chemically synthesized and, in contrast to tag-specific capture probes (such as Nickel-NTA or Glutathione), they can be used for purification of proteins free of genetic modifications (such as His or GST tags). Despite these attractive features of aptamers as capture probes, there are only a few reports on aptamer-based protein purification and none of them includes a test of the purified protein's activity, thus, leaving discouraging doubts about method's ability to purify proteins in their active state. The goal of this work was to prove that aptamers could facilitate isolation of active proteins. We refined a complete aptamer-based affinity purification procedure, which takes 4 h to complete. We further applied this procedure to purify two recombinant proteins, MutS and AlkB, from bacterial cell culture: 0.21 mg of 85%-pure AlkB from 4 mL of culture and 0.24 mg of 82%-pure MutS from 0.5 mL of culture. Finally, we proved protein activity by two capillary electrophoresis based assays: an enzymatic assay for AlkB and a DNA-binding assay for MutS. We suggest that in combination with aptamer selection for non-purified protein targets in crude cell lysate, aptamer-based purification provides a means of fast isolation of tag-free recombinant proteins in their native state without the use of antibodies. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. In situ dissection of RNA functional subunits by domain-specific chromatin isolation by RNA purification (dChIRP).

    PubMed

    Quinn, Jeffrey J; Chang, Howard Y

    2015-01-01

    Here we describe domain-specific chromatin isolation by RNA purification (dChIRP), a technique for dissecting the functional domains of a target RNA in situ. For an RNA of interest, dChIRP can identify domain-level intramolecular and intermolecular RNA-RNA, RNA-protein, and RNA-DNA interactions and maps the RNA's genomic binding sites with higher precision than domain-agnostic methods. We illustrate how this technique has been applied to the roX1 lncRNA to resolve its domain-level architecture, discover its protein- and chromatin-interacting domains, and map its occupancy on the X chromosome.

  8. Two-step ion-exchange chromatographic purification combined with reversed-phase chromatography to isolate C-peptide for mass spectrometric analysis.

    PubMed

    Kabytaev, Kuanysh; Durairaj, Anita; Shin, Dmitriy; Rohlfing, Curt L; Connolly, Shawn; Little, Randie R; Stoyanov, Alexander V

    2016-02-01

    A liquid chromatography with mass spectrometry on-line platform that includes the orthogonal techniques of ion exchange and reversed phase chromatography is applied for C-peptide analysis. Additional improvement is achieved by the subsequent application of cation- and anion-exchange purification steps that allow for isolating components that have their isoelectric points in a narrow pH range before final reversed-phase mass spectrometry analysis. The utility of this approach for isolating fractions in the desired "pI window" for profiling complex mixtures is discussed. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Ultrasensitive Electrochemical Detection of mRNA Using Branched DNA Amplifiers

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Mao, Xun; Liu, Guodong; Wang, Shengfu

    2008-11-01

    We describe here an ultrasensitive electrochemical detection of m RNA protocol without RNA purification and PCR amplification. The new m RNA electrical detection capability is coupled to the amplification feature of branched DNA (bDNA) technology and with the nagnetic beads based electrochemical bioassay.

  10. Creative Uses for Calculator-based Laboratory (CBL) Technology in Chemistry.

    ERIC Educational Resources Information Center

    Sales, Cynthia L.; Ragan, Nicole M.; Murphy, Maureen Kendrick

    1999-01-01

    Reviews three projects that use a graphing calculator linked to a calculator-based laboratory device as a portable data-collection system for students in chemistry classes. Projects include Isolation, Purification and Quantification of Buckminsterfullerene from Woodstove Ashes; Determination of the Activation Energy Associated with the…

  11. Small Laccase from "Streptomyces Coelicolor"--An Ideal Model Protein/Enzyme for Undergraduate Laboratory Experience

    ERIC Educational Resources Information Center

    Cook, Ryan; Hannon, Drew; Southard, Jonathan N.; Majumdar, Sudipta

    2018-01-01

    A one semester undergraduate biochemistry laboratory experience is described for an understanding of recombinant technology from gene cloning to protein characterization. An integrated experimental design includes three sequential modules: molecular cloning, protein expression and purification, and protein analysis and characterization. Students…

  12. Desalination and Water Purification Technology Roadmap

    DTIC Science & Technology

    2003-01-01

    those contaminants to re-enter the water cycle in the near-term). Developing safe disposal options will require, among other activities, research on...destroyed. The global water cycle dictates that the water we use today has been used countless times before, and will be used countless times again

  13. Application of partition technology to particle electrophoresis

    NASA Technical Reports Server (NTRS)

    Van Alstine, James M.; Harris, J. Milton; Karr, Laurel J.; Bamberger, Stephan; Matsos, Helen C.; Snyder, Robert S.

    1989-01-01

    The effects of polymer-ligand concentration on particle electrophoretic mobility and partition in aqueous polymer two-phase systems are investigated. Polymer coating chemistry and affinity ligand synthesis, purification, and analysis are conducted. It is observed that poly (ethylene glycol)-ligands are effective for controlling particle electrophoretic mobility.

  14. Secretory immunoglobulin purification from whey by chromatographic techniques.

    PubMed

    Matlschweiger, Alexander; Engelmaier, Hannah; Himmler, Gottfried; Hahn, Rainer

    2017-08-15

    Secretory immunoglobulins (SIg) are a major fraction of the mucosal immune system and represent potential drug candidates. So far, platform technologies for their purification do not exist. SIg from animal whey was used as a model to develop a simple, efficient and potentially generic chromatographic purification process. Several chromatographic stationary phases were tested. A combination of two anion-exchange steps resulted in the highest purity. The key step was the use of a small-porous anion exchanger operated in flow-through mode. Diffusion of SIg into the resin particles was significantly hindered, while the main impurities, IgG and serum albumin, were bound. In this step, initial purity was increased from 66% to 89% with a step yield of 88%. In a second anion-exchange step using giga-porous material, SIg was captured and purified by step or linear gradient elution to obtain fractions with purities >95%. For the step gradient elution step yield of highly pure SIg was 54%. Elution of SIgA and SIgM with a linear gradient resulted in a step yield of 56% and 35%, respectively. Overall yields for both anion exchange steps were 43% for the combination of flow-through and step elution mode. Combination of flow-through and linear gradient elution mode resulted in a yield of 44% for SIgA and 39% for SIgM. The proposed process allows the purification of biologically active SIg from animal whey in preparative scale. For future applications, the process can easily be adopted for purification of recombinant secretory immunoglobulin species. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Industrial Membrane Filtration and Short-bed Fractal Separation Systems for Separating Monomers from Heterogeneous Plant Material

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kearney, M; Kochergin, V; Hess, R

    2005-03-31

    Large-scale displacement of petroleum will come from low-cost cellulosic feedstocks such as straw and corn stover crop residues. This project has taken a step toward making this projection a reality by reducing capital and energy costs, the two largest cost factors associated with converting cellulosic biomass to chemicals and fuels. The technology exists for using acid or enzyme hydrolysis processes to convert biomass feedstock (i.e., waste cellulose such as straw, corn stover, and wood) into their base monomeric sugar building blocks, which can, in turn, be processed into chemicals and fuels using a number of innovative fermentation technologies. However, whilemore » these processes are technically possible, practical and economic barriers make these processes only marginally feasible or not feasible at all. These barriers are due in part to the complexity and large fixed and recurring capital costs of unit operations including filtration, chromatographic separation, and ion exchange. This project was designed to help remove these barriers by developing and implementing new purification and separation technologies that will reduce the capital costs of the purification and chromatographic separation units by 50% to 70%. The technologies fundamental to these improvements are: (a) highly efficient clarification and purification systems that use screening and membrane filtration to eliminate suspended solids and colloidal material from feed streams and (b) fractal technology based chromatographic separation and ion exchange systems that can substitute for conventional systems but at much smaller size and cost. A non-hazardous ''raw sugar beet juice'' stream (75 to 100 gal/min) was used for prototype testing of these technologies. This raw beet juice stream from the Amalgamated Sugar LLC plant in Twin Falls, Idaho contained abrasive materials and membrane foulants. Its characteristics were representative of an industrial-scale heterogeneous plant extract/hydrolysis stream, and therefore was an ideal model system for developing new separation equipment. Subsequent testing used both synthetic acid hydrolysate and corn stover derived weak acid hydrolysate (NREL produced). A two-phased approach was used for the research and development described in this project. The first level of study involved testing the new concepts at the bench level. The bench-scale evaluations provided fundamental understanding of the processes, building and testing small prototype systems, and determining the efficiency of the novel processes. The second level of study, macro-level, required building larger systems that directly simulated industrial operations and provided validation of performance to minimize financial risk during commercialization. The project goals and scope included: (1) Development of low-capital alternatives to conventional crop-based purification/separation processes; and (2) Development of each process to the point that transition to commercial operation is low risk. The project reporting period was January 2001 to December 2004. This included a one year extension of the project (without additional funding).« less

  16. Extraction and downstream processing of plant-derived recombinant proteins.

    PubMed

    Buyel, J F; Twyman, R M; Fischer, R

    2015-11-01

    Plants offer the tantalizing prospect of low-cost automated manufacturing processes for biopharmaceutical proteins, but several challenges must be addressed before such goals are realized and the most significant hurdles are found during downstream processing (DSP). In contrast to the standardized microbial and mammalian cell platforms embraced by the biopharmaceutical industry, there are many different plant-based expression systems vying for attention, and those with the greatest potential to provide inexpensive biopharmaceuticals are also the ones with the most significant drawbacks in terms of DSP. This is because the most scalable plant systems are based on the expression of intracellular proteins in whole plants. The plant tissue must therefore be disrupted to extract the product, challenging the initial DSP steps with an unusually high load of both particulate and soluble contaminants. DSP platform technologies can accelerate and simplify process development, including centrifugation, filtration, flocculation, and integrated methods that combine solid-liquid separation, purification and concentration, such as aqueous two-phase separation systems. Protein tags can also facilitate these DSP steps, but they are difficult to transfer to a commercial environment and more generic, flexible and scalable strategies to separate target and host cell proteins are preferable, such as membrane technologies and heat/pH precipitation. In this context, clarified plant extracts behave similarly to the feed stream from microbes or mammalian cells and the corresponding purification methods can be applied, as long as they are adapted for plant-specific soluble contaminants such as the superabundant protein RuBisCO. Plant-derived pharmaceutical proteins cannot yet compete directly with established platforms but they are beginning to penetrate niche markets that allow the beneficial properties of plants to be exploited, such as the ability to produce 'biobetters' with tailored glycans, the ability to scale up production rapidly for emergency responses and the ability to produce commodity recombinant proteins on an agricultural scale. Copyright © 2015 Elsevier Inc. All rights reserved.

  17. Dodecyl Maltopyranoside Enabled Purification of Active Human GABA Type A Receptors for Deep and Direct Proteomic Sequencing*

    PubMed Central

    Zhang, Xi; Miller, Keith W.

    2015-01-01

    The challenge in high-quality membrane proteomics is all about sample preparation prior to HPLC, and the cell-to-protein step poses a long-standing bottleneck. Traditional protein extraction methods apply ionic or poly-disperse detergents, harsh denaturation, and repeated protein/peptide precipitation/resolubilization afterward, but suffer low yield, low reproducibility, and low sequence coverage. Contrary to attempts to subdue, we resolved this challenge by providing proteins nature-and-activity-promoting conditions throughout preparation. Using 285-kDa hetero-pentameric human GABA type A receptor overexpressed in HEK293 as a model, we describe a n-dodecyl-β-d-maltopyranoside/cholesteryl hemisuccinate (DDM/CHS)-based affinity purification method, that produced active receptors, supported protease activity, and allowed high performance with both in-gel and direct gel-free proteomic analyses—without detergent removal. Unlike conventional belief that detergents must be removed before HPLC MS, the high-purity low-dose nonionic detergent DDM did not interfere with peptides, and obviated removal or desalting. Sonication or dropwise addition of detergent robustly solubilized over 90% of membrane pellets. The purification conditions were comparable to those applied in successful crystallizations of most membrane proteins. These results enabled streamlined proteomics of human synaptic membrane proteins, and more importantly, allowed directly coupling proteomics with crystallography to characterize both static and dynamic structures of membrane proteins in crystallization pipelines. PMID:25473089

  18. Isolation of centromeric-tandem repetitive DNA sequences by chromatin affinity purification using a HaloTag7-fused centromere-specific histone H3 in tobacco.

    PubMed

    Nagaki, Kiyotaka; Shibata, Fukashi; Kanatani, Asaka; Kashihara, Kazunari; Murata, Minoru

    2012-04-01

    The centromere is a multi-functional complex comprising centromeric DNA and a number of proteins. To isolate unidentified centromeric DNA sequences, centromere-specific histone H3 variants (CENH3) and chromatin immunoprecipitation (ChIP) have been utilized in some plant species. However, anti-CENH3 antibody for ChIP must be raised in each species because of its species specificity. Production of the antibodies is time-consuming and costly, and it is not easy to produce ChIP-grade antibodies. In this study, we applied a HaloTag7-based chromatin affinity purification system to isolate centromeric DNA sequences in tobacco. This system required no specific antibody, and made it possible to apply a highly stringent wash to remove contaminated DNA. As a result, we succeeded in isolating five tandem repetitive DNA sequences in addition to the centromeric retrotransposons that were previously identified by ChIP. Three of the tandem repeats were centromere-specific sequences located on different chromosomes. These results confirm the validity of the HaloTag7-based chromatin affinity purification system as an alternative method to ChIP for isolating unknown centromeric DNA sequences. The discovery of more than two chromosome-specific centromeric DNA sequences indicates the mosaic structure of tobacco centromeres. © Springer-Verlag 2011

  19. Cost-effective Expression and Purification of Antimicrobial and Host Defense Peptides in Escherichia coli

    PubMed Central

    Bommarius, B.; Jenssen, H.; Elliott, M.; Kindrachuk, J.; Pasupuleti, Mukesh; Gieren, H; Jaeger, K.-E.; Hancock, R.E. W.

    2010-01-01

    Cationic antimicrobial host defense peptides (HDPs) combat infection by directly killing a wide variety of microbes, and/or modulating host immunity. HDPs have great therapeutic potential against antibiotic-resistant bacteria, viruses and even parasites, but there are substantial roadblocks to their therapeutic application. High manufacturing costs associated with amino acid precursors have limited the delivery of inexpensive therapeutics through industrial-scale chemical synthesis. Conversely, the production of peptides in bacteria by recombinant DNA technology has been impeded by the antimicrobial activity of these peptides and their susceptibility to proteolytic degradation, while subsequent purification of recombinant peptides often requires multiple steps and has not been cost-effective. Here we have developed methodologies appropriate for large-scale industrial production of HDPs; in particular, we describe (i) a method, using fusions to SUMO, for producing high yields of intact recombinant HDPs in bacteria without significant toxicity; and (ii) a simplified 2-step purification method appropriate for industrial use. We have used this method to produce seven HDPs to date (IDR1, MX226, LL37, CRAMP, HHC-10, E5 and E6). Using this technology, pilot-scale fermentation (10 L) was performed to produce large quantities of biologically active cationic peptides. Together, these data indicate that this new method represents a cost-effective means to enable commercial enterprises to produce HDPs in large-scale under Good Laboratory Manufacturing Practice (GMP) conditions for therapeutic application in humans. PMID:20713107

  20. A simple electroelution method for rapid protein purification: isolation and antibody production of alpha toxin from Clostridium septicum.

    PubMed

    Vázquez-Iglesias, Lorena; Estefanell-Ucha, Borja; Barcia-Castro, Leticia; Páez de la Cadena, María; Álvarez-Chaver, Paula; Ayude-Vázquez, Daniel; Rodríguez-Berrocal, Francisco Javier

    2017-01-01

    Clostridium septicum produces a number of diseases in human and farm animals which, in most of the cases, are fatal without clinical intervention. Alpha toxin is an important agent and the unique lethal virulent factor produced by Clostridium septicum. This toxin is haemolytic, highly lethal and necrotizing activities but is being used as an antigen to develop animal vaccines. The aim of this study was to isolate the alpha toxin of Clostridium septicum and produce highly specific antibodies against it. In this work, we have developed a simple and efficient method for alpha toxin purification, based on electroelution that can be used as a time-saving method for purifying proteins. This technique avoids contamination by other proteins that could appear during other protein purification techniques such chromatography. The highly purified toxin was used to produce polyclonal antibodies. The specificity of the antibodies was tested by western blot and these antibodies can be applied to the quantitative determination of alpha toxin by slot blot.

  1. A simple electroelution method for rapid protein purification: isolation and antibody production of alpha toxin from Clostridium septicum

    PubMed Central

    Estefanell-Ucha, Borja; Barcia-Castro, Leticia; Páez de la Cadena, María; Álvarez-Chaver, Paula; Ayude-Vázquez, Daniel; Rodríguez-Berrocal, Francisco Javier

    2017-01-01

    Clostridium septicum produces a number of diseases in human and farm animals which, in most of the cases, are fatal without clinical intervention. Alpha toxin is an important agent and the unique lethal virulent factor produced by Clostridium septicum. This toxin is haemolytic, highly lethal and necrotizing activities but is being used as an antigen to develop animal vaccines. The aim of this study was to isolate the alpha toxin of Clostridium septicum and produce highly specific antibodies against it. In this work, we have developed a simple and efficient method for alpha toxin purification, based on electroelution that can be used as a time-saving method for purifying proteins. This technique avoids contamination by other proteins that could appear during other protein purification techniques such chromatography. The highly purified toxin was used to produce polyclonal antibodies. The specificity of the antibodies was tested by western blot and these antibodies can be applied to the quantitative determination of alpha toxin by slot blot. PMID:28652930

  2. Adsorption of Selected Antibiotics to Resins in Extracorporeal Blood Purification.

    PubMed

    Harm, Stephan; Gruber, Anna; Gabor, Franz; Hartmann, Jens

    2016-01-01

    Extracorporeal blood purification systems (EBS) use specific adsorbents for the elimination of toxins and cytokines. The aim of this study was to test different adsorbents for their ability to reduce antibiotics in parallel to extracorporeal blood purification therapy. The in vitro adsorption experiments were carried out in human plasma with a newly established hydrophobic resin (Amberchrom CG161c) and adsorbents commercially available and approved in the clinics. The concentration of antibiotic was chosen equivalent to the recommended therapeutic dosage applied intravenously and was measured in plasma using ELISA test kits and high-performance liquid chromatography methods. The adsorbent that reduced all tested antibiotics in plasma close to the detection limit was the dia MARS AC250, which is an activated charcoal involved in the Molecular Adsorbents Recirculation System. For better antibiotic monitoring in sepsis treatment, further investigations have to be performed to determine the clearance rate of antibiotics by different EBS devices. © 2015 S. Karger AG, Basel.

  3. Entanglement of purification through holographic duality

    NASA Astrophysics Data System (ADS)

    Umemoto, Koji; Takayanagi, Tadashi

    2018-06-01

    The gauge/gravity correspondence discovered two decades ago has had a profound influence on how the basic laws in physics should be formulated. In spite of the predictive power of holographic approaches (for example, when they are applied to strongly coupled condensed-matter physics problems), the fundamental reasons behind their success remain unclear. Recently, the role of quantum entanglement has come to the fore. Here we explore a quantity that connects gravity and quantum information in the light of the gauge/gravity correspondence. This is given by the minimal cross-section of the entanglement wedge that connects two disjoint subsystems in a gravity dual. In particular, we focus on various inequalities that are satisfied by this quantity. They suggest that it is a holographic counterpart of the quantity called entanglement of purification, which measures a bipartite correlation in a given mixed state. We give a heuristic argument that supports this identification based on a tensor network interpretation of holography. This predicts that the entanglement of purification satisfies the strong superadditivity for holographic conformal field theories.

  4. Poly(N-Isopropylacrylamide)-co-Acrylamide Hydrogels for the Controlled Release of Bromelain from Agroindustrial Residues of Ananas comosus.

    PubMed

    Croisfelt, Fernanda; Martins, Bianca C; Rescolino, Robson; Coelho, Diego F; Zanchetta, Beatriz; Mazzola, Priscila G; Goulart, Luis Ricardo; Pessoa, Adalberto; Tambourgi, Elias B; Silveira, Edgar

    2015-12-01

    This works reports the purification of bromelain extracted from Ananas comosus industrial residues by ethanol purification, its partial characterization from the crude extract as well as the ethanol purified enzyme, and its application onto poly(N-isopropylacrylamide)-co-acrylamide hydrogels. Bromelain was recovered within the 30-70 % ethanol fraction, which achieved a purification factor of 3.12-fold, and yielded more than 90 % of its initial activity. The resulting purified bromelain contained more than 360 U · mg(-1), with a maximum working temperature of 60 °C and pH of 8.0. Poly(N-isopropylacrylamide)-co-acrylamide hydrogels presented a swelling rate of 125 %, which was capable of loading 56 % of bromelain from the solution, and was able to release up to 91 % of the retained bromelain. Ethanol precipitation is suitable for bromelain recovery and application onto poly(N-isopropylacrylamide)-co-acrylamide hydrogels based on its processing time and the applied ethanol prices. Georg Thieme Verlag KG Stuttgart · New York.

  5. An "in Silico" DNA Cloning Experiment for the Biochemistry Laboratory

    ERIC Educational Resources Information Center

    Elkins, Kelly M.

    2011-01-01

    This laboratory exercise introduces students to concepts in recombinant DNA technology while accommodating a major semester project in protein purification, structure, and function in a biochemistry laboratory for junior- and senior-level undergraduate students. It is also suitable for forensic science courses focused in DNA biology and advanced…

  6. Middle School and pH?

    ERIC Educational Resources Information Center

    Herricks, Susan

    2007-01-01

    A local middle school requested that the Water Center of Advanced Materials for Purification of Water With Systems (WaterCAMPWS), a National Science Foundation Science and Technology Center, provide an introduction to pH for their seventh-grade water-based service learning class. After sorting through a multitude of information about pH, a…

  7. Current status and future prospects for enabling chemistry technology in the drug discovery process.

    PubMed

    Djuric, Stevan W; Hutchins, Charles W; Talaty, Nari N

    2016-01-01

    This review covers recent advances in the implementation of enabling chemistry technologies into the drug discovery process. Areas covered include parallel synthesis chemistry, high-throughput experimentation, automated synthesis and purification methods, flow chemistry methodology including photochemistry, electrochemistry, and the handling of "dangerous" reagents. Also featured are advances in the "computer-assisted drug design" area and the expanding application of novel mass spectrometry-based techniques to a wide range of drug discovery activities.

  8. The growth of materials processing in space - A history of government support for new technology

    NASA Technical Reports Server (NTRS)

    Mckannan, E. C.

    1983-01-01

    Development of a given technology for national defense and large systems developments when the task is too large or risky for entrepreneurs, yet is clearly in the best interest of the nation are discussed. Advanced research to identify areas of interest was completed. Examples of commercial opportunities are the McDonnell-Douglas Corporation purification process for pharmaceutical products and the Microgravity Research Associates process for growing gallium arsenide crystals in space.

  9. Synthesis and purification of galacto-oligosaccharides: state of the art.

    PubMed

    Vera, Carlos; Córdova, Andrés; Aburto, Carla; Guerrero, Cecilia; Suárez, Sebastián; Illanes, Andrés

    2016-12-01

    Lactose-derived non-digestible oligosaccharides are prominent components of functional foods. Among them, galacto-oligosaccharides (GOS) outstand for being prebiotics whose health-promoting effects are supported on strong scientific evidences, having unique properties as substitutes of human milk oligosaccharides in formulas for newborns and infants. GOS are currently produced enzymatically in a kinetically-controlled reaction of lactose transgalactosylation catalyzed by β-galactosidases from different microbial strains. The enzymatic synthesis of GOS, although being an established technology, still offers many technological challenges and opportunities for further development that has to be considered within the framework of functional foods which is the most rapidly expanding market within the food sector. This paper presents the current technological status of GOS production, its main achievements and challenges. Most of the problems yet to be solved refer to the rather low GOS yields attainable that rarely exceed 40 %, corresponding to lactose conversions around 60 %. This means that the product or reaction (raw GOS) contains significant amounts of residual lactose and monosaccharides (glucose and galactose). Efforts to increase such yields have been for the most part unsuccessful, even though improvements by genetic and protein engineering strategies are to be expected in the near future. Low yields impose a burden on downstream processing to obtain a GOS product of the required purity. Different strategies for raw GOS purification are reviewed and their technological significance is appraised.

  10. Purification and characterization of two isoenzymes of lipoxygenase from soybeans.

    PubMed

    Diel, E; Stan, H J

    1978-01-01

    A chromatographic procedure for the purification of two lipoxygenase isoenzymes (linoleate: O2 oxidoreductase, EC 1.13.11.12.) from soybean is described. The procedure for the purification of isoenzyme L-1 includes optimalized extraction, ammonium sulfate fractionation, heat treatment and gradient elution from a CM-Sephadex C-50 column. The purification of L-2 includes ammonium sulfate fractionation, gelfiltration on Sephadex G-150 and gradient elution from a DEAE-cellulose column. Both isoenzymes L-1 and L-2 appear homogeneous after Disc-PAGE. The isoelectric points are 5.6 for L-1 and 5.8 for L-2. Molecular weights are estimated as 100,000 for L-1 as well as L-2 applying three different methods. Both isoenzymes contain 0.9 mol iron per mol protien. The estimated turn over numbers are 8,200 mol linoleate per mol enzyme and min for L-1 and 3,100 for L-2. Amino acid compositions determined after acid hydrolysis show marked differences between L-1 and L-2, particularly with respect to the amino acids Lys, Phe, Ser, Gly and Leu. L-1 posesses a total of 9 cysteine molecules, 6 of which are present as disulfide bonds. L-2 posesses a total of 8 cysteine molecules with only one disulfide bond.

  11. Isolation of mitochondria from Saccharomyces cerevisiae using magnetic bead affinity purification

    PubMed Central

    Liao, Pin-Chao; Boldogh, Istvan R.; Siegmund, Stephanie E.

    2018-01-01

    Isolated mitochondria are widely used to study the function of the organelle. Typically, mitochondria are prepared using differential centrifugation alone or in conjunction with density gradient ultracentrifugation. However, mitochondria isolated using differential centrifugation contain membrane or organelle contaminants, and further purification of crude mitochondria by density gradient ultracentrifugation requires large amounts of starting material, and is time-consuming. Mitochondria have also been isolated by irreversible binding to antibody-coated magnetic beads. We developed a method to prepare mitochondria from budding yeast that overcomes many of the limitations of other methods. Mitochondria are tagged by insertion of 6 histidines (6xHis) into the TOM70 (Translocase of outer membrane 70) gene at its chromosomal locus, isolated using Ni-NTA (nickel (II) nitrilotriacetic acid) paramagnetic beads and released from the magnetic beads by washing with imidazole. Mitochondria prepared using this method contain fewer contaminants, and are similar in ultrastructure as well as protein import and cytochrome c oxidase complex activity compared to mitochondria isolated by differential centrifugation. Moreover, this isolation method is amenable to small samples, faster than purification by differential and density gradient centrifugation, and more cost-effective than purification using antibody-coated magnetic beads. Importantly, this method can be applied to any cell type where the genetic modification can be introduced by CRISPR or other methods. PMID:29698455

  12. A novel method for separation of caseins from milk by phosphates precipitation.

    PubMed

    Yen, Chon-Ho; Lin, Yin-Shen; Tu, Ching-Fu

    2015-01-01

    Milk protein of farm animals is difficult to isolate because of the presence of casein micelles, which are hard to separate from whey by using centrifugation or filtration. Insoluble casein micelles also create an obstacle for purification instruments to operate efficiently. The conventional method, to precipitate caseins by lowering pH to 4.6 and then recover the whey fraction for further purification using chromatography techniques, is not applicable to proteins having an isoelectric point similar to caseins. In addition, the acid condition used for casein removal usually leads to significantly poor yields and reduced biological activities. In this study, a novel method of precipitating caseins under neutral or weak acidic conditions is presented. The method employs a phosphate salt and a freeze-thaw procedure to obtain a casein-free whey protein fraction. This fraction contains more than 90% yield with little loss of bioactivity of the target protein, and is readily available for further chromatographic purification. This method was successfully applied to purify recombinant human factor IX and recombinant hirudin from the milk of transgenic pigs in the presented study. It is an efficient pretreatment approach prior to chromatographic purification of milk protein from farm animals and particularly of great value to collect those recombinants secreted from transgenic livestock.

  13. Carbon Nanotubes for Human Space Flight

    NASA Technical Reports Server (NTRS)

    Scott, Carl D.; Files, Brad; Yowell, Leonard

    2003-01-01

    Single-wall carbon nanotubes offer the promise of a new class of revolutionary materials for space applications. The Carbon Nanotube Project at NASA Johnson Space Center has been actively researching this new technology by investigating nanotube production methods (arc, laser, and HiPCO) and gaining a comprehensive understanding of raw and purified material using a wide range of characterization techniques. After production and purification, single wall carbon nanotubes are processed into composites for the enhancement of mechanical, electrical, and thermal properties. This "cradle-to-grave" approach to nanotube composites has given our team unique insights into the impact of post-production processing and dispersion on the resulting material properties. We are applying our experience and lessons-learned to developing new approaches toward nanotube material characterization, structural composite fabrication, and are also making advances in developing thermal management materials and electrically conductive materials in various polymer-nanotube systems. Some initial work has also been conducted with the goal of using carbon nanotubes in the creation of new ceramic materials for high temperature applications in thermal protection systems. Human space flight applications such as advanced life support and fuel cell technologies are also being investigated. This discussion will focus on the variety of applications under investigation.

  14. Plasma-based water purification: Challenges and prospects for the future

    NASA Astrophysics Data System (ADS)

    Foster, John E.

    2017-05-01

    Freshwater scarcity derived from seasonal weather variations, climate change, and over-development has led to serious consideration for water reuse. Water reuse involves the direct processing of wastewater for either indirect or directly potable water reuse. In either case, advanced water treatment technologies will be required to process the water to the point that it can be reused in a meaningful way. Additionally, there is growing concern regarding micropollutants, such as pharmaceuticals and personal care products, which have been detected in finished drinking water not removed by conventional means. The health impact of these contaminants in low concentration is not well understood. Pending regulatory action, the removal of these contaminants by water treatment plants will also require advanced technology. One new and emerging technology that could potentially address the removal of micropollutants in both finished drinking water as well as wastewater slated for reuse is plasma-based water purification. Plasma in contact with liquid water generates a host of reactive species that attack and ultimately mineralize contaminants in solution. This interaction takes place in the boundary layer or interaction zone centered at the plasma-liquid water interface. An understanding of the physical processes taking place at the interface, though poorly understood, is key to the optimization of plasma-based water purifiers. High electric field conditions, large density gradients, plasma-driven chemistries, and fluid dynamic effects prevail in this multiphase region. The region is also the source function for longer-lived reactive species that ultimately treat the water. Here, we review the need for advanced water treatment methods and in the process, make the case for plasma-based methods. Additionally, we survey the basic methods of interacting plasma with liquid water (including a discussion of breakdown processes in water), the current state of understanding of the physical processes taking place at the plasma-liquid interface, and the role these processes play in water purification. The development of plasma diagnostics usable in this multiphase environment along with modeling efforts aimed at elucidating physical processes taking place at the interface are also detailed. Key experiments that demonstrate the capability of plasma-based water treatment are also reviewed. The technical challenges to the implementation of plasma-based water reactors are also discussed. We conclude with a discussion of prospects for the future of plasma-based water purification.

  15. Fluorescent IgG fusion proteins made in E. coli

    PubMed Central

    Luria, Yael; Raichlin, Dina; Benhar, Itai

    2012-01-01

    Antibodies are among the most powerful tools in biological and biomedical research and are presently the fastest growing category of new bio-pharmaceutics. The most common format of antibody applied for therapeutic, diagnostic and analytical purposes is the IgG format. For medical applications, recombinant IgGs are made in cultured mammalian cells in a process that is too expensive to be considered for producing antibodies for diagnostic and analytical purposes. Therefore, for such purposes, mouse monoclonal antibodies or polyclonal sera from immunized animals are used. While looking for an easier and more rapid way to prepare full-length IgGs for therapeutic purposes, we recently developed and reported an expression and purification protocol for full-length IgGs, and IgG-based fusion proteins in E. coli, called “Inclonals.” By applying the Inclonals technology, we could generate full-length IgGs that are genetically fused to toxins. The aim of the study described herein was to evaluate the possibility of applying the “Inclonals” technology for preparing IgG-fluorophore fusion proteins. We found that IgG fused to the green fluorescent proteins enhanced GFP (EGFP) while maintaining functionality in binding, lost most of its fluorescence during the refolding process. In contrast, we found that green fluorescent Superfolder GFP (SFGFP)-fused IgG and red fluorescent mCherry-fused IgG were functional in antigen binding and maintained fluorescence intensity. In addition, we found that we can link several SFGFPs in tandem to each IgG, with fluorescence intensity increasing accordingly. Fluorescent IgGs made in E. coli may become attractive alternatives to monoclonal or polyclonal fluorescent antibodies derived from animals. PMID:22531449

  16. Multiple functions of caprylic acid-induced impurity precipitation for process intensification in monoclonal antibody purification.

    PubMed

    Trapp, Anja; Faude, Alexander; Hörold, Natalie; Schubert, Sven; Faust, Sabine; Grob, Thilo; Schmidt, Stefan

    2018-05-02

    New emerging technologies delivering benefits in terms of process robustness and economy are an inevitable prerequisite for monoclonal antibody purification processes intensification. Caprylic acid was proven as an effective precipitating agent enabling efficient precipitaton of product- and process-related impurities while leaving the antibody in solution. This purification step at mild acidic pH was therefore introduced in generic antibody platform approaches after Protein A capture and evaluated for its impact regarding process robustness and antibody stability. Comparison of 13 different monoclonal antibodies showed significant differences in antibody recovery between 65-95% during caprylic acid-induced impurity precipitation. Among six compared physicochemical properties, isoelectric point of the antibody domains was figured out to correlate with yield. Antibodies with mild acidic pI of the light chain were significantly susceptible to caprylic acid-induced precipitation resulting in lower yields. Virus clearance studies revealed that caprylic acid provided complete virus inactivation of an enveloped virus. Multiple process relevant factors such as pH range, caprylic acid concentration and antibody stability were investigated in this study to enable an intensified purification process including caprylic acid precipitation for HCP removal of up to 2 log 10 reduction values at mAb yields >90% while also contributing to the virus safety of the process. Copyright © 2018 Elsevier B.V. All rights reserved.

  17. Role of messenger RNA-ribosome complex in complementary DNA display.

    PubMed

    Naimuddin, Mohammed; Ohtsuka, Isao; Kitamura, Koichiro; Kudou, Motonori; Kimura, Shinnosuke

    2013-07-15

    In vitro display technologies such as ribosome display and messenger RNA (mRNA)/complementary DNA (cDNA) display are powerful methods that can generate library diversities on the order of 10(10-14). However, in mRNA and cDNA display methods, the end use diversity is two orders of magnitude lower than initial diversity and is dependent on the downstream processes that act as limiting factors. We found that in our previous cDNA display protocol, the purification of protein fusions by the use of streptavidin matrices from cell-free translation mixtures had poor efficiency (∼10-15%) that seriously affected the diversity of the purified library. Here, we have investigated and optimized the protocols that provided remarkable purification efficiencies. The stalled ribosome in the mRNA-ribosome complex was found to impede this purification efficiency. Among the various conditions tested, destabilization of ribosomes by appropriate concentration of metal chelating agents in combination with an optimal temperature of 30°C were found to be crucial and effective for nearly complete isolation of protein fusions from the cell-free translation system. Thus, this protocol provided 8- to 10-fold increased efficiency of purification over the previous method and results in retaining the diversity of the library by approximately an order of magnitude-important for directed evolution. We also discuss the possible effects in the fabrication of protein chips. Copyright © 2013 Elsevier Inc. All rights reserved.

  18. Coupled plasma filtration adsorption: rationale, technical development and early clinical experience.

    PubMed

    Ronco, Claudio; Brendolan, Alessandra; d'Intini, Vincenzo; Ricci, Zaccaria; Wratten, Mary Lou; Bellomo, Rinaldo

    2003-01-01

    The adjuvant treatment of sepsis remains a major therapeutic challenge. Blood purification is theoretically appealing if the humoral theory of sepsis is accepted as the basis for intervention. In this setting, blood purification would provide a broad-based restoration of humoral homeostasis thereby avoiding both excessive inflammation and counterinflammation. Several techniques of blood purification have been tried or are under active investigation. One of these is the so-called coupled plasma filtration adsorption (CPFA). CPFA is a novel extracorporeal blood purification therapy aimed at nonselectively reducing the circulating levels and activities of both pro- and anti-inflammatory mediators during sepsis and multiorgan failure. In vitro studies have shown CPFA to be effective in binding a broad range of such mediators proving its technical efficacy. Subsequent animal models have shown a beneficial effect on survival in endotoxemia. These studies have provided the necessary technical developments and biologic rationale for initial human studies. Two phase I/IIa clinical studies have now been performed. Both studies have shown that CPFA improves blood pressure and restores immune function in patients with severe sepsis and multiorgan dysfunction. In this article, we will discuss some of the basic principles involved in sorbent technology, and how these may contribute to treatment efficacy, review animal experiments with CPFA and finally discuss the results of recent human studies and their implications. Copyright 2003 S. Karger AG, Basel

  19. Magnetic bead purification of labeled DNA fragments forhigh-throughput capillary electrophoresis sequencing

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Elkin, Christopher; Kapur, Hitesh; Smith, Troy

    2001-09-15

    We have developed an automated purification method for terminator sequencing products based on a magnetic bead technology. This 384-well protocol generates labeled DNA fragments that are essentially free of contaminates for less than $0.005 per reaction. In comparison to laborious ethanol precipitation protocols, this method increases the phred20 read length by forty bases with various DNA templates such as PCR fragments, Plasmids, Cosmids and RCA products. Our method eliminates centrifugation and is compatible with both the MegaBACE 1000 and ABIPrism 3700 capillary instruments. As of September 2001, this method has produced over 1.6 million samples with 93 percent averaging 620more » phred20 bases as part of Joint Genome Institutes Production Process.« less

  20. Design and manufacture of monoclonal antibodies for radioimmunotherapy.

    PubMed

    Hale, G; Berrie, E; Bird, P

    2004-12-01

    antibodies is fundamental to their use for radioimmunotherapy. Besides the right selection of antibody specificity and affinity, recombinant antibodies can be designed to simplify manufacture and minimise unwanted side effects. Although many innovative new technologies have been developed in recent years, antibodies are still most commonly produced from mammalian cells and purified by column chromatography. Purification methods have to be designed and validated to remove potential contaminants, especially retroviruses, which in principle might be present in mammalian cell lines. Adherence to relevant ''Good Manufacturing Practices'' is mandatory in the production of any medicinal product and there are numerous guidelines regarding the manufacture of antibodies. This article outlines some methods used for fermentation, purification and quality control of antibodies intended for radiolabelling.

  1. Solvent-resistant nanofiltration for product purification and catalyst recovery in click chemistry reactions.

    PubMed

    Cano-Odena, Angels; Vandezande, Pieter; Fournier, David; Van Camp, Wim; Du Prez, Filip E; Vankelecom, Ivo F J

    2010-01-18

    The quickly developing field of "click" chemistry would undoubtedly benefit from the availability of an easy and efficient technology for product purification to reduce the potential health risks associated with the presence of copper in the final product. Therefore, solvent-resistant nanofiltration (SRNF) membranes have been developed to selectively separate "clicked" polymers from the copper catalyst and solvent. By using these solvent-stable cross-linked polyimide membranes in diafiltration, up to 98 % of the initially present copper could be removed through the membrane together with the DMF solvent, the polymer product being almost completely retained. This paper also presents the first SRNF application in which the catalyst permeates through the membrane and the reaction product is retained.

  2. Membranes with Surface-Enhanced Antifouling Properties for Water Purification

    PubMed Central

    Shahkaramipour, Nima; Tran, Thien N.; Ramanan, Sankara; Lin, Haiqing

    2017-01-01

    Membrane technology has emerged as an attractive approach for water purification, while mitigation of fouling is key to lower membrane operating costs. This article reviews various materials with antifouling properties that can be coated or grafted onto the membrane surface to improve the antifouling properties of the membranes and thus, retain high water permeance. These materials can be separated into three categories, hydrophilic materials, such as poly(ethylene glycol), polydopamine and zwitterions, hydrophobic materials, such as fluoropolymers, and amphiphilic materials. The states of water in these materials and the mechanisms for the antifouling properties are discussed. The corresponding approaches to coat or graft these materials on the membrane surface are reviewed, and the materials with promising performance are highlighted. PMID:28273869

  3. In situ synthesis of protein arrays.

    PubMed

    He, Mingyue; Stoevesandt, Oda; Taussig, Michael J

    2008-02-01

    In situ or on-chip protein array methods use cell free expression systems to produce proteins directly onto an immobilising surface from co-distributed or pre-arrayed DNA or RNA, enabling protein arrays to be created on demand. These methods address three issues in protein array technology: (i) efficient protein expression and availability, (ii) functional protein immobilisation and purification in a single step and (iii) protein on-chip stability over time. By simultaneously expressing and immobilising many proteins in parallel on the chip surface, the laborious and often costly processes of DNA cloning, expression and separate protein purification are avoided. Recently employed methods reviewed are PISA (protein in situ array) and NAPPA (nucleic acid programmable protein array) from DNA and puromycin-mediated immobilisation from mRNA.

  4. Purification Techniques of Bacteriocins from Lactic Acid Bacteria and Other Gram-Positive Bacteria

    NASA Astrophysics Data System (ADS)

    Saavedra, Lucila; Sesma, Fernando

    The search for new antimicrobial peptides produced by lactic acid ­bacteria and other Gram-positive microorganisms has become an interesting field of research in the past decades. The fact that bacteriocins are active against numerous foodborne and human pathogens, are produced by generally regarded as safe (GRAS) microorganisms, and are readily degraded by proteolytic host systems makes them attractive candidates for biotechnological applications. However, before suggesting or choosing a new bacteriocin for future technology developments, it is necessary to elucidate its biochemical structure and its mode of action, which may be carried out once the bacteriocin is purified to homogeneity. This chapter focuses on describing the main strategies used for the purification of numerous bacteriocins.

  5. Membranes with Surface-Enhanced Antifouling Properties for Water Purification.

    PubMed

    Shahkaramipour, Nima; Tran, Thien N; Ramanan, Sankara; Lin, Haiqing

    2017-03-05

    Membrane technology has emerged as an attractive approach for water purification, while mitigation of fouling is key to lower membrane operating costs. This article reviews various materials with antifouling properties that can be coated or grafted onto the membrane surface to improve the antifouling properties of the membranes and thus, retain high water permeance. These materials can be separated into three categories, hydrophilic materials, such as poly(ethylene glycol), polydopamine and zwitterions, hydrophobic materials, such as fluoropolymers, and amphiphilic materials. The states of water in these materials and the mechanisms for the antifouling properties are discussed. The corresponding approaches to coat or graft these materials on the membrane surface are reviewed, and the materials with promising performance are highlighted.

  6. Data in support of three phase partitioning of zingibain, a milk-clotting enzyme from Zingiber officinale Roscoe rhizomes

    PubMed Central

    Gagaoua, Mohammed; Hafid, Kahina; Hoggas, Naouel

    2016-01-01

    This paper describes data related to a research article titled “Three Phase Partitioning of zingibain, a milk-clotting enzyme from Zingiber officinale Roscoe rhizomes” (Gagaoua et al., 2015) [1]. Zingibain (EC 3.4.22.67), is a coagulant cysteine protease and a meat tenderizer agent that have been reported to produce satisfactory final products in dairy and meat technology, respectively. Zingibains were exclusively purified using chromatographic techniques with very low yield purification. This paper includes data of the effect of temperature, usual salts and organic solvents on the efficiency of the three phase partitioning (TPP) system. Also it includes data of the kinetic activity characterization of the purified zingibain using TPP purification approach. PMID:26909379

  7. Efficient sortase-mediated N-terminal labeling of TEV protease cleaved recombinant proteins.

    PubMed

    Sarpong, Kwabena; Bose, Ron

    2017-03-15

    A major challenge in attaching fluorophores or other handles to proteins is the availability of a site-specific labeling strategy that provides stoichiometric modification without compromising protein integrity. We developed a simple approach that combines TEV protease cleavage, sortase modification and affinity purification to N-terminally label proteins. To achieve stoichiometrically-labeled protein, we included a short affinity tag in the fluorophore-containing peptide for post-labeling purification of the modified protein. This strategy can be easily applied to any recombinant protein with a TEV site and we demonstrate this on Epidermal Growth Factor Receptor (EGFR) and Membrane Scaffold Protein (MSP) constructs. Copyright © 2017 Elsevier Inc. All rights reserved.

  8. Refined separation of combined Fe–Hf from rock matrices for isotope analyses using AG-MP-1M and Ln-Spec chromatographic extraction resins

    PubMed Central

    Cheng, Ting; Nebel, Oliver; Sossi, Paolo A.; Chen, Fukun

    2014-01-01

    A combined procedure for separating Fe and Hf from a single rock digestion is presented. In a two-stage chromatographic extraction process, a purified Fe fraction is first quantitatively separated from the rock matrix using AG-MP-1M resin in HCl. Hafnium is subsequently isolated using a modified version of a commonly applied method using Eichrom LN-Spec resin. Our combined method includes:•Purification of Fe from the rock matrix using HCl, ready for mass spectrometric analysis.•Direct loading of the matrix onto the resin that is used for Hf purification.•Collection of a Fe-free Hf fraction. PMID:26150946

  9. The effect of column purification on cDNA indirect labelling for microarrays

    PubMed Central

    Molas, M Lia; Kiss, John Z

    2007-01-01

    Background The success of the microarray reproducibility is dependent upon the performance of standardized procedures. Since the introduction of microarray technology for the analysis of global gene expression, reproducibility of results among different laboratories has been a major problem. Two of the main contributors to this variability are the use of different microarray platforms and different laboratory practices. In this paper, we address the latter question in terms of how variation in one of the steps of a labelling procedure affects the cDNA product prior to microarray hybridization. Results We used a standard procedure to label cDNA for microarray hybridization and employed different types of column chromatography for cDNA purification. After purifying labelled cDNA, we used the Agilent 2100 Bioanalyzer and agarose gel electrophoresis to assess the quality of the labelled cDNA before its hybridization onto a microarray platform. There were major differences in the cDNA profile (i.e. cDNA fragment lengths and abundance) as a result of using four different columns for purification. In addition, different columns have different efficiencies to remove rRNA contamination. This study indicates that the appropriate column to use in this type of protocol has to be experimentally determined. Finally, we present new evidence establishing the importance of testing the method of purification used during an indirect labelling procedure. Our results confirm the importance of assessing the quality of the sample in the labelling procedure prior to hybridization onto a microarray platform. Conclusion Standardization of column purification systems to be used in labelling procedures will improve the reproducibility of microarray results among different laboratories. In addition, implementation of a quality control check point of the labelled samples prior to microarray hybridization will prevent hybridizing a poor quality sample to expensive micorarrays. PMID:17597522

  10. The effect of column purification on cDNA indirect labelling for microarrays.

    PubMed

    Molas, M Lia; Kiss, John Z

    2007-06-27

    The success of the microarray reproducibility is dependent upon the performance of standardized procedures. Since the introduction of microarray technology for the analysis of global gene expression, reproducibility of results among different laboratories has been a major problem. Two of the main contributors to this variability are the use of different microarray platforms and different laboratory practices. In this paper, we address the latter question in terms of how variation in one of the steps of a labelling procedure affects the cDNA product prior to microarray hybridization. We used a standard procedure to label cDNA for microarray hybridization and employed different types of column chromatography for cDNA purification. After purifying labelled cDNA, we used the Agilent 2100 Bioanalyzer and agarose gel electrophoresis to assess the quality of the labelled cDNA before its hybridization onto a microarray platform. There were major differences in the cDNA profile (i.e. cDNA fragment lengths and abundance) as a result of using four different columns for purification. In addition, different columns have different efficiencies to remove rRNA contamination. This study indicates that the appropriate column to use in this type of protocol has to be experimentally determined. Finally, we present new evidence establishing the importance of testing the method of purification used during an indirect labelling procedure. Our results confirm the importance of assessing the quality of the sample in the labelling procedure prior to hybridization onto a microarray platform. Standardization of column purification systems to be used in labelling procedures will improve the reproducibility of microarray results among different laboratories. In addition, implementation of a quality control check point of the labelled samples prior to microarray hybridization will prevent hybridizing a poor quality sample to expensive micorarrays.

  11. Topic model-based mass spectrometric data analysis in cancer biomarker discovery studies.

    PubMed

    Wang, Minkun; Tsai, Tsung-Heng; Di Poto, Cristina; Ferrarini, Alessia; Yu, Guoqiang; Ressom, Habtom W

    2016-08-18

    A fundamental challenge in quantitation of biomolecules for cancer biomarker discovery is owing to the heterogeneous nature of human biospecimens. Although this issue has been a subject of discussion in cancer genomic studies, it has not yet been rigorously investigated in mass spectrometry based proteomic and metabolomic studies. Purification of mass spectometric data is highly desired prior to subsequent analysis, e.g., quantitative comparison of the abundance of biomolecules in biological samples. We investigated topic models to computationally analyze mass spectrometric data considering both integrated peak intensities and scan-level features, i.e., extracted ion chromatograms (EICs). Probabilistic generative models enable flexible representation in data structure and infer sample-specific pure resources. Scan-level modeling helps alleviate information loss during data preprocessing. We evaluated the capability of the proposed models in capturing mixture proportions of contaminants and cancer profiles on LC-MS based serum proteomic and GC-MS based tissue metabolomic datasets acquired from patients with hepatocellular carcinoma (HCC) and liver cirrhosis as well as synthetic data we generated based on the serum proteomic data. The results we obtained by analysis of the synthetic data demonstrated that both intensity-level and scan-level purification models can accurately infer the mixture proportions and the underlying true cancerous sources with small average error ratios (<7 %) between estimation and ground truth. By applying the topic model-based purification to mass spectrometric data, we found more proteins and metabolites with significant changes between HCC cases and cirrhotic controls. Candidate biomarkers selected after purification yielded biologically meaningful pathway analysis results and improved disease discrimination power in terms of the area under ROC curve compared to the results found prior to purification. We investigated topic model-based inference methods to computationally address the heterogeneity issue in samples analyzed by LC/GC-MS. We observed that incorporation of scan-level features have the potential to lead to more accurate purification results by alleviating the loss in information as a result of integrating peaks. We believe cancer biomarker discovery studies that use mass spectrometric analysis of human biospecimens can greatly benefit from topic model-based purification of the data prior to statistical and pathway analyses.

  12. Current status and future prospects for enabling chemistry technology in the drug discovery process

    PubMed Central

    Djuric, Stevan W.; Hutchins, Charles W.; Talaty, Nari N.

    2016-01-01

    This review covers recent advances in the implementation of enabling chemistry technologies into the drug discovery process. Areas covered include parallel synthesis chemistry, high-throughput experimentation, automated synthesis and purification methods, flow chemistry methodology including photochemistry, electrochemistry, and the handling of “dangerous” reagents. Also featured are advances in the “computer-assisted drug design” area and the expanding application of novel mass spectrometry-based techniques to a wide range of drug discovery activities. PMID:27781094

  13. Potential means of support for materials processing in space. A history of government support for new technology

    NASA Technical Reports Server (NTRS)

    Mckannan, E. C.

    1983-01-01

    Development of a given technology for national defense and large systems developments when the task is too large or risky for entrepreneurs, yet is clearly in the best interest of the nation are discussed. Advanced research to identify areas of interest was completed. Examples of commercial opportunities are the McDonnell-Douglas Corporation purification process for pharmaceutical products and the Microgravity Research Associates process for growing gallium arsenide crystals in space.

  14. Application of Mathematical and Three-Dimensional Computer Modeling Tools in the Planning of Processes of Fuel and Energy Complexes

    NASA Astrophysics Data System (ADS)

    Aksenova, Olesya; Nikolaeva, Evgenia; Cehlár, Michal

    2017-11-01

    This work aims to investigate the effectiveness of mathematical and three-dimensional computer modeling tools in the planning of processes of fuel and energy complexes at the planning and design phase of a thermal power plant (TPP). A solution for purification of gas emissions at the design development phase of waste treatment systems is proposed employing mathematical and three-dimensional computer modeling - using the E-nets apparatus and the development of a 3D model of the future gas emission purification system. Which allows to visualize the designed result, to select and scientifically prove economically feasible technology, as well as to ensure the high environmental and social effect of the developed waste treatment system. The authors present results of a treatment of planned technological processes and the system for purifying gas emissions in terms of E-nets. using mathematical modeling in the Simulink application. What allowed to create a model of a device from the library of standard blocks and to perform calculations. A three-dimensional model of a system for purifying gas emissions has been constructed. It allows to visualize technological processes and compare them with the theoretical calculations at the design phase of a TPP and. if necessary, make adjustments.

  15. Recent advances in enzyme extraction strategies: A comprehensive review.

    PubMed

    Nadar, Shamraja S; Pawar, Rohini G; Rathod, Virendra K

    2017-08-01

    The increasing interest of industrial enzymes demands for development of new downstream strategies for maximizing enzyme recovery. The significant efforts have been focused on the development of newly adapted technologies to purify enzymes in catalytically active form. Recently, an aqueous two phase system (ATPS) is emerged as powerful tools for efficient extraction and purification of enzymes due to their versatility, lower cost, process integration capability and easy scale-up. The present review gives an overview of effect of parameters such as tie line length, pH, neutral salts, properties of polymer and salt involved in traditional polymer/polymer and polymer/salt ATPS for enzyme recovery. Further, advanced ATPS have been developed based on alcohols, surfactants, micellar compounds to avoid tedious recovery steps for getting desired enzyme. In order to improve the selectivity and efficiency of ATPS, recent approaches of conventional ATPS combined with different techniques like affinity ligands, ionic liquids, thermoseparating polymers and microfluidic device based ATPS have been reviewed. Moreover, three phase partitioning is also highlighted for enzymes enrichment as a blooming technology for efficiently integrated bioseparation techniques. At the end, it includes an overview of CLEAs technology and organic-inorganic nanoflowers preparation as novel strategies for simultaneous extraction, purification and immobilization of enzymes. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Generation of Murine Monoclonal Antibodies by Hybridoma Technology.

    PubMed

    Holzlöhner, Pamela; Hanack, Katja

    2017-01-02

    Monoclonal antibodies are universal binding molecules and are widely used in biomedicine and research. Nevertheless, the generation of these binding molecules is time-consuming and laborious due to the complicated handling and lack of alternatives. The aim of this protocol is to provide one standard method for the generation of monoclonal antibodies using hybridoma technology. This technology combines two steps. Step 1 is an appropriate immunization of the animal and step 2 is the fusion of B lymphocytes with immortal myeloma cells in order to generate hybrids possessing both parental functions, such as the production of antibody molecules and immortality. The generated hybridoma cells were then recloned and diluted to obtain stable monoclonal cell cultures secreting the desired monoclonal antibody in the culture supernatant. The supernatants were tested in enzyme-linked immunosorbent assays (ELISA) for antigen specificity. After the selection of appropriate cell clones, the cells were transferred to mass cultivation in order to produce the desired antibody molecule in large amounts. The purification of the antibodies is routinely performed by affinity chromatography. After purification, the antibody molecule can be characterized and validated for the final test application. The whole process takes 8 to 12 months of development, and there is a high risk that the antibody will not work in the desired test system.

  17. Technology: New Ways for Clean Water

    ERIC Educational Resources Information Center

    Roberts, Amanda S.

    2012-01-01

    Water purification promotes healthy living. While the developing world is working to provide its citizens with future access to clean water sources, the demand for that water is a pressing need today. It should be understood that drinking water, sanitation, and hygiene are interwoven and are all necessary for the overall improved standard of…

  18. Profiling and quantitative evaluation of three Nickel-Coated magnetic matrices for purification of recombinant proteins: lelpful hints for the optimized nanomagnetisable matrix preparation

    PubMed Central

    2011-01-01

    Background Several materials are available in the market that work on the principle of protein magnetic fishing by their histidine (His) tags. Little information is available on their performance and it is often quoted that greatly improved purification of histidine-tagged proteins from crude extracts could be achieved. While some commercial magnetic matrices could be used successfully for purification of several His-tagged proteins, there are some which have been proved to operate just for a few extent of His-tagged proteins. Here, we address quantitative evaluation of three commercially available Nickel nanomagnetic beads for purification of two His-tagged proteins expressed in Escherichia coli and present helpful hints for optimized purification of such proteins and preparation of nanomagnetisable matrices. Results Marked differences in the performance of nanomagnetic matrices, principally on the basis of their specific binding capacity, recovery profile, the amount of imidazole needed for protein elution and the extent of target protein loss and purity were obtained. Based on the aforesaid criteria, one of these materials featured the best purification results (SiMAG/N-NTA/Nickel) for both proteins at the concentration of 4 mg/ml, while the other two (SiMAC-Nickel and SiMAG/CS-NTA/Nickel) did not work well with respect to specific binding capacity and recovery profile. Conclusions Taken together, functionality of different types of nanomagnetic matrices vary considerably. This variability may not only be dependent upon the structure and surface chemistry of the matrix which in turn determine the affinity of interaction, but, is also influenced to a lesser extent by the physical properties of the protein itself. Although the results of the present study may not be fully applied for all nanomagnetic matrices, but provide a framework which could be used to profiling and quantitative evaluation of other magnetisable matrices and also provide helpful hints for those researchers facing same challenge. PMID:21824404

  19. Design of functional guanidinium ionic liquid aqueous two-phase systems for the efficient purification of protein.

    PubMed

    Ding, Xueqin; Wang, Yuzhi; Zeng, Qun; Chen, Jing; Huang, Yanhua; Xu, Kaijia

    2014-03-07

    A series of novel cationic functional hexaalkylguanidinium ionic liquids and anionic functional tetraalkylguanidinium ionic liquids have been devised and synthesized based on 1,1,3,3-tetramethylguanidine. The structures of the ionic liquids (ILs) were confirmed by (1)H nuclear magnetic resonance ((1)H NMR) and 13C nuclear magnetic resonance (13C NMR) and the production yields were all above 90%. Functional guanidinium ionic liquid aqueous two-phase systems (FGIL-ATPSs) have been first designed with these functional guanidinium ILs and phosphate solution for the purification of protein. After phase separation, proteins had transferred into the IL-rich phase and the concentrations of proteins were determined by measuring the absorbance at 278 nm using an ultra violet visible (UV-vis) spectrophotometer. The advantages of FGIL-ATPSs were compared with ordinary ionic liquid aqueous two-phase systems (IL-ATPSs). The proposed FGIL-ATPS has been applied to purify lysozyme, trypsin, ovalbumin and bovine serum albumin. Single factor experiments were used to research the effects of the process, such as the amount of ionic liquid (IL), the concentration of salt solution, temperature and the amount of protein. The purification efficiency reaches to 97.05%. The secondary structure of protein during the experimental process was observed upon investigation using UV-vis spectrophotometer, Fourier-transform infrared spectroscopy (FT-IR) and circular dichroism spectrum (CD spectrum). The precision, stability and repeatability of the process were investigated. The mechanisms of purification were researched by dynamic light scattering (DLS), determination of the conductivity and transmission electron microscopy (TEM). It was suggested that aggregation and embrace phenomenon play a significant role in the purification of proteins. All the results show that FGIL-ATPSs have huge potential to offer new possibility in the purification of proteins. Copyright © 2014 Elsevier B.V. All rights reserved.

  20. A RAPID DNA EXTRACTION METHOD FOR PCR IDENTIFICATION OF FUNGAL INDOOR AIR CONTAMINANTS

    EPA Science Inventory

    Following air sampling, fungal DNA needs to be extracted and purified to a state suitable for laboratory use. Our laboratory has developed a simple method of extraction and purification of fungal DNA appropriate for enzymatic manipulation and polymerase chain reaction (PCR) appli...

  1. An ammonium sulfate/ethanol aqueous two-phase system combined with ultrasonication for the separation and purification of lithospermic acid B from Salvia miltiorrhiza Bunge.

    PubMed

    Guo, Y X; Han, J; Zhang, D Y; Wang, L H; Zhou, L L

    2012-07-01

    We studied the effect of ultrasonication extraction technology combined with ammonium sulfate/ethanol aqueous two-phase system (ATPS) for the separation of lithospermic acid B (LAB) from Salvia miltiorrhiza Bunge. According to the literature and preliminary studies, ammonium sulfate concentration, ethanol concentration, pH, ultrasonication power, ultrasonication time and the ratio of solvent-to-solid were investigated using a single factor design to identify the factors affecting separation. Taking into consideration a simultaneous increase in LAB recovery (R (%)) and partition coefficient (K), the best performance of the ATPS was obtained at 25°C and pH 2 using ammonium sulfate 22% (w/w) and ethanol 30% (w/w). To keep the solvent-to-solid ratio at 10, response surface methodology was used to find the optimal ultrasonication power and ultrasonication time. Quadratic models were predicted for LAB yield in the upper phase. Optimal conditions of 572.1 W ultrasonication power and 42.2 min produced a maximum yield of LAB of 42.16 mg g(-1) sample. There was no obvious degradation of LAB with ultrasound under the applied conditions, and the experimental yield of LAB was 42.49 mg g(-1) sample and the purity was 55.28% (w/w), which was much higher than that obtained using conventional extraction. The present study demonstrated that ultrasound coupled with aqueous two-phase systems is very efficient tool for the extraction and purification of LAB from Salvia miltiorrhiza Bunge. Copyright © 2011 Elsevier B.V. All rights reserved.

  2. Gas purification in the dense phase at the CATS terminal

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Openshaw, P.J.; Carnell, P.J.H.; Rhodes, E.F.

    The purification and transportation of natural gas at very high pressures can help to minimize the capital cost of pipelines and processing equipment. However, complex mixtures of hydrocarbons undergo unusual phase changes, such as retrograde condensation, as the temperature and pressure are altered. The Central Area Transmission System (CATS) is a joint venture of Amoci, BG, Amerada Hess, Phillips, Agip and Fina operated by Amoco on behalf of the owners. The design of the CATS terminal has provided an interesting processing challenge. The terminal receives a total of 1.6 Bscf/d of rich gas from a number of offshore fields. Allmore » are relatively sweet but the small amounts of H{sub 2}S and Hg are removed. Fixed bed technology was selected as the most economic purification process, while minimizing hydrocarbon loss and operator involvement. Conventionally, the raw gas would be split into the different hydrocarbon fractions and each would be processed separately. This would require the installation of a large number of reactors. A more elegant solution is to treat the gas on arrival at the terminal in the dense phase. This option raised questions around whether a fixed bed would be prone to fouling, could the pressure drop be kept low enough to avoid phase separation and would inadvertent wetting by condensation cause problems. Details are given of the test work carried out to prove the viability of using fixed bed technology for dense phase gas processing, the eventual design adopted and the performance over the first year of service.« less

  3. Response surface methodology to optimize partition and purification of two recombinant oxidoreductase enzymes, glucose dehydrogenase and d-galactose dehydrogenase in aqueous two-phase systems.

    PubMed

    Shahbaz Mohammadi, Hamid; Mostafavi, Seyede Samaneh; Soleimani, Saeideh; Bozorgian, Sajad; Pooraskari, Maryam; Kianmehr, Anvarsadat

    2015-04-01

    Oxidoreductases are an important family of enzymes that are used in many biotechnological processes. An experimental design was applied to optimize partition and purification of two recombinant oxidoreductases, glucose dehydrogenase (GDH) from Bacillus subtilis and d-galactose dehydrogenase (GalDH) from Pseudomonas fluorescens AK92 in aqueous two-phase systems (ATPS). Response surface methodology (RSM) with a central composite rotatable design (CCRD) was performed to optimize critical factors like polyethylene glycol (PEG) concentration, concentration of salt and pH value. The best partitioning conditions was achieved in an ATPS composed of 12% PEG-6000, 15% K2HPO4 with pH 7.5 at 25°C, which ensured partition coefficient (KE) of 66.6 and 45.7 for GDH and GalDH, respectively. Under these experimental conditions, the activity of GDH and GalDH was 569.5U/ml and 673.7U/ml, respectively. It was found that these enzymes preferentially partitioned into the top PEG-rich phase and appeared as single bands on SDS-PAGE gel. Meanwhile the validity of the response model was confirmed by a good agreement between predicted and experimental results. Collectively, according to the obtained data it can be inferred that the ATPS optimization using RSM approach can be applied for recovery and purification of any enzyme from oxidoreductase family. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. Integration of Gas Chromatography Mass Spectrometry Methods for Differentiating Ricin Preparation Methods

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Wunschel, David S.; Melville, Angela M.; Ehrhardt, Christopher J.

    2012-05-17

    The investigation of crimes involving chemical or biological agents is infrequent, but presents unique analytical challenges. The protein toxin ricin is encountered more frequently than other agents and is found in the seeds of the castor plant Ricinus communis. Typically, the toxin is extracted from castor seeds utilizing a variety of different recipes that result in varying purity of the toxin. Moreover, these various purification steps can also leave or differentially remove a variety of exogenous and endogenous residual components with the toxin that may indicate the type and number of purification steps involved. We have applied three gas chromatographicmore » - mass spectrometric (GC-MS) based analytical methods to measure the variation in seed carbohydrates and castor oil ricinoleic acid as well as the presence of solvents used for purification. These methods were applied to the same samples prepared using four previously identified toxin preparation methods starting from four varieties of castor seeds. The individual data sets for seed carbohydrate profiles, ricinoleic acid or acetone amount each provided information capable of differentiating different types of toxin preparations across seed types. However, the integration of the data sets using multivariate factor analysis provided a clear distinction of all samples based on the preparation method and independent of the seed source. In particular the abundance of mannose, arabinose, fucose, ricinoleic acid and acetone were shown to be important differentiating factors. These complementary tools provide a more confident determination of the method of toxin preparation.« less

  5. Isolation and purification of food-grade C-phycocyanin from Arthrospira platensis and its determination in confectionery by HPLC with diode array detection.

    PubMed

    Kissoudi, Maria; Sarakatsianos, Ioannis; Samanidou, Victoria

    2018-02-01

    C-Phycocyanin is the major phycobiliprotein in Arthrospira platensis, also known as Spirulina, which is a cyanobacterium used as a dietary supplement because of its powerful effects on body and brain. C-phycocyanin is a blue-colored accessory photosynthetic pigment with multiple applications in food industry as natural dye or additive, and in pharmaceuticals. This study presents a simple protocol for the extraction and purification of food-grade C-phycocyanin from Arthrospira platensis. The cell lysis of cyanobacterium was performed by sonication combined with repeated freezing and thawing cycles. The purification of the crude extract of C-phycocyanin was carried out by ammonium sulfate precipitation followed by ion exchange chromatography resulting in 2.5 purity. The purity of phycocyanobilin chromophore has been tested by UV-visible spectrophotometry by monitoring the absorption after each stage of purification. A high-performance liquid chromatography method has been developed and validated for the determination of food-grade C-phycocyanin. Intra- and interday precision values less than 5.6% and recovery greater than 91.2% indicated high precision and accuracy of the method for analysis of C-phycocyanin. The method has been applied to commercial confectionery of blue color and to the purified protein obtained in the first stage of the study. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Defluoridation technology for drinking water and tea by green synthesized Fe3O4/Al2O3 nanoparticles coated polyurethane foams for rural communities.

    PubMed

    Kumari, Sonu; Khan, Suphiya

    2017-08-14

    Fluoride (F) contaminated ground water poses a serious public health concern to rural population with unaffordable purification technologies. Therefore, development of a cost-effective, portable, environment and user-friendly defluoridation technique is imperative. In the present study, we report on the development of a green and cost-effective method that utilizes Fe 3 O 4 and Al 2 O 3 nanoparticles (NPs) that were synthesized using jojoba defatted meal. These NPs were impregnated on to polyurethane foam (PUF) and made into tea infusion bags. The Al 2 O 3 NPs-PUF displayed a higher water defluoridation capacity of 43.47 mg g -1 of F as compared to 34.48 mg g -1 of F with Fe 3 O 4 NPs-PUF. The synthesized Al 2 O 3 -PUF infusion bags removed the F that was under the permissible limit of 1.5 mg L -1 . The sorption experiments were conducted to verify the effect of different parameters such as pH, contact time, size of PUF and initial F concentration. The different properties of adsorbent were characterized using a combination of FESEM, EDX, XRD and FTIR techniques, respectively. The calculated total cost per NPs-PUF pouch developed is as low as US $0.05, which makes the technology most suitable for rural communities. This paper will be beneficial for researchers working toward further improvement in water purification technologies.

  7. 40 CFR 63.5610 - What definitions apply to this subpart?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... from a railcar using nitrogen or water displacement and storage of carbon disulfide in a storage vessel using nitrogen or water padding. Cellophane means a thin, transparent cellulose material, which is... chloride, or chloroacetic acid, to produce a particular cellulose ether; (iii) Washing and purification of...

  8. 40 CFR 63.5610 - What definitions apply to this subpart?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... from a railcar using nitrogen or water displacement and storage of carbon disulfide in a storage vessel using nitrogen or water padding. Cellophane means a thin, transparent cellulose material, which is... chloride, or chloroacetic acid, to produce a particular cellulose ether; (iii) Washing and purification of...

  9. 40 CFR 63.5610 - What definitions apply to this subpart?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... from a railcar using nitrogen or water displacement and storage of carbon disulfide in a storage vessel using nitrogen or water padding. Cellophane means a thin, transparent cellulose material, which is... chloride, or chloroacetic acid, to produce a particular cellulose ether; (iii) Washing and purification of...

  10. Novel tiO2 nanocatalysts for wastewater purification: tapping energy from the sun.

    PubMed

    Liu, Y; Li, J; Qiu, X; Burda, C

    2006-01-01

    Water treatment using TiO2 semiconductor as a durable heterogeneous photocatalyst has been the focus of environmentalists in recent years. Currently, we developed an inexpensive and highly efficient approach for synthesizing nitrogen-doped TiO2 with lower band-gap energy that can respond to visible light. Doping on the molecular scale led to an enhanced nitrogen concentration of up to 21.8%. Reflectance measurements showed the synthesized N-doped TiO2 nanoparticles are catalytically active with the absorbance that extends into the visible region up to 600 nm. The water purification potential of this new class of compound was evaluated by studying the photodegradation of Acid Orange 7 (AO7) and E. coli. Experiments were conducted to compare the photocatalytic activities of N-doped TiO2 nanocatalysts and commercially available Degussa P25 power under identical solar light exposure. N-doped TiO2 demonstrated superior photocatalytic activities in both chemical compound degradation and bactericidal reactions. The result of this study shows the potential of applying new generations of catalyst for wastewater purification and disinfection.

  11. Early process development of API applied to poorly water-soluble TBID.

    PubMed

    Meise, Marius; Niggemann, Matthias; Dunens, Alexandra; Schoenitz, Martin; Kuschnerow, Jan C; Kunick, Conrad; Scholl, Stephan

    2018-05-01

    Finding and optimising of synthesis processes for active pharmaceutical ingredients (API) is time consuming. In the finding phase, established methods for synthesis, purification and formulation are used to achieve a high purity API for biological studies. For promising API candidates, this is followed by pre-clinical and clinical studies requiring sufficient quantities of the active component. Ideally, these should be produced with a process representative for a later production process and suitable for scaling to production capacity. This work presents an overview of different approaches for process synthesis based on an existing lab protocol. This is demonstrated for the production of the model drug 4,5,6,7-tetrabromo-2-(1H-imidazol-2-yl) isoindolin-1,3-dione (TBID). Early batch synthesis and purification procedures typically suffer from low and fluctuating yields and purities due to poor process control. In a first step the literature synthesis and purification procedure was modified and optimized using solubility measurements, targeting easier and safer processing for consecutive studies. Copyright © 2018 Elsevier B.V. All rights reserved.

  12. Purification of Derivatized Oligosaccharides by Solid Phase Extraction for Glycomic Analysis

    PubMed Central

    Zhang, Qiwei; Li, Henghui; Feng, Xiaojun; Liu, Bi-Feng; Liu, Xin

    2014-01-01

    Profiling of glycans released from proteins is very complex and important. To enhance the detection sensitivity, chemical derivatization is required for the analysis of carbohydrates. Due to the interference of excess reagents, a simple and reliable purification method is usually necessary for the derivatized oligosaccharides. Various SPE based methods have been applied for the clean-up process. To demonstrate the differences among these methods, seven types of self-packed SPE cartridges were systematically compared in this study. The optimized conditions were determined for each type of cartridge and it was found that microcrystalline cellulose was the most appropriate SPE material for the purification of derivatized oligosaccharide. Normal phase HPLC analysis of the derivatized maltoheptaose was realized with a detection limit of 0.12 pmol (S N−1 = 3) and a recovery over 70%. With the optimized SPE method, relative quantification analysis of N-glycans from model glycoproteins were carried out accurately and over 40 N-glycans from human serum samples were determined regardless of the isomers. Due to the high stability and sensitivity, microcrystalline cellulose cartridge showed potential applications in glycomics analysis. PMID:24705408

  13. Use of T-2 toxin-immobilized amine-activated beads as an efficient affinity purification matrix for the isolation of specific IgY.

    PubMed

    Edupuganti, Soujanya Ratna; Edupuganti, Om Prakash; O'Kennedy, Richard; Defrancq, Eric; Boullanger, Stéphanie

    2013-04-01

    An affinity purification method that isolates T-2 toxin-specific IgY utilizing a T-2-toxin-immobilized column was developed. The T-2 toxin was covalently coupled via a carbonyldiimidazole-activated hydroxyl functional group to amine-activated sepharose beads. The affinity-purified IgY was characterized by gel electrophoresis, fast protein liquid chromatography, enzyme-linked immunosorbant assay, surface plasmon resonance and mass spectrometry. A competitive inhibition ELISA (CI-ELISA) was performed using affinity-purified IgY with a T-2 toxin detection sensitivity of 30 ng/mL, which falls within the maximum permissible limit of 100 ng/mL. The cross reactivity of IgY towards deoxynivalenol, zearalenone, fumonisin B1 and HT-2 was significantly reduced after affinity purification. A surface plasmon resonance (SPR)-based inhibition assay was also applied for quantitative determination of T-2 toxin in spiked wheat samples. The results obtained indicate the feasibility of utilizing this IgY-based assay for the detection of T-2 toxin in food samples.

  14. Determination of the Self-organized Mesoporous Materials Sorption Capacity in Water Treatment Processes Using Mössbauer Spectroscopy

    NASA Astrophysics Data System (ADS)

    Feklistov, D. Yu.; Filippov, V. P.; Kurchatov, I. M.; Laguntsov, N. I.

    Different approaches to water purification are considered. It is shown that for developing of effective purification methods in the different water treatment technologies special knowledge is necessary. This knowledge deals with physical-chemical processes of used reagents interaction with contaminating impurities. The obtaining of the necessary data on physics-chemical processes such as the formation of chemical compounds of iron, the change in valence state of iron, adsorption and absorption pollutants are analyzed. The usage of iron compounds as a cleaners and the water treatment from the iron pollutant are the events, in which Mössbauer spectroscopy allows to determine not only the chemical changes, but also to obtain quantitative data on the reaction products and on the sizes of reaction products and their quantities.

  15. Tandem dye-ligand chromatography and biospecific elution applied to the purification of glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides.

    PubMed Central

    Hey, Y; Dean, P D

    1983-01-01

    1. A total of 65 immobilized triazine dyes were screened for their ability to purify the dual-nucleotide-specific glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides. From this screen a 'negative' (Matrex Gel Purple A) and a 'positive' (Matrex Gel Orange B) adsorbent were found to be the best in terms of overall purification and yield and were therefore combined to give the best purification. 2. Glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides was purified approx. 56-fold in a two-step tandem chromatographic system using Matrex Gel Purple A followed by Matrex Gel Orange B chromatography to a specific activity of 228 units/mg of protein in a final yield of 73%. 3. A study of the elution characteristics of glucose-6-phosphate dehydrogenase bound to Matrex Gel Orange B by KCl (pulse and gradient) and biospecific eluents (pulse) was carried out. NADP+, NADPH and adenosine 2',5'-bisphosphate were found to be the only effective biospecific eluents. A pulse of 50 microM-NADP+ (1/2 column vol.) was found to give a better purification than a 0-1 M-KCl gradient and therefore was the preferred method of elution. 4. Presaturation of the enzyme with various nucleotides was carried out to determine the effect on the subsequent binding of glucose-6-phosphate dehydrogenase to Matrex Gel Orange B. The results of these and biospecific-elution studies lead us to propose two possible schemes to explain the mechanism of the dye-protein interaction. 5. Reusability, capacity of the adsorbent and effect of varying the ligand concentration were also studied in the purification of glucose-6-phosphate dehydrogenase on Matrex Gel Orange B. Images Fig. 1. PMID:6847623

  16. Tandem dye-ligand chromatography and biospecific elution applied to the purification of glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides.

    PubMed

    Hey, Y; Dean, P D

    1983-02-01

    1. A total of 65 immobilized triazine dyes were screened for their ability to purify the dual-nucleotide-specific glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides. From this screen a 'negative' (Matrex Gel Purple A) and a 'positive' (Matrex Gel Orange B) adsorbent were found to be the best in terms of overall purification and yield and were therefore combined to give the best purification. 2. Glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides was purified approx. 56-fold in a two-step tandem chromatographic system using Matrex Gel Purple A followed by Matrex Gel Orange B chromatography to a specific activity of 228 units/mg of protein in a final yield of 73%. 3. A study of the elution characteristics of glucose-6-phosphate dehydrogenase bound to Matrex Gel Orange B by KCl (pulse and gradient) and biospecific eluents (pulse) was carried out. NADP+, NADPH and adenosine 2',5'-bisphosphate were found to be the only effective biospecific eluents. A pulse of 50 microM-NADP+ (1/2 column vol.) was found to give a better purification than a 0-1 M-KCl gradient and therefore was the preferred method of elution. 4. Presaturation of the enzyme with various nucleotides was carried out to determine the effect on the subsequent binding of glucose-6-phosphate dehydrogenase to Matrex Gel Orange B. The results of these and biospecific-elution studies lead us to propose two possible schemes to explain the mechanism of the dye-protein interaction. 5. Reusability, capacity of the adsorbent and effect of varying the ligand concentration were also studied in the purification of glucose-6-phosphate dehydrogenase on Matrex Gel Orange B.

  17. Dynamic pH junction high-speed counter-current chromatography coupled with microwave-assisted extraction for online separation and purification of alkaloids from Stephania cepharantha.

    PubMed

    Yuan, Zhiquan; Xiao, Xiaohua; Li, Gongke

    2013-11-22

    A simple and efficient dynamic pH junction high-speed counter-current chromatography method was developed and further applied to the online extraction, separation and purification of alkaloids from Stephania cepharantha by coupling with microwave-assisted extraction. Mineral acid and organic base were added into the mobile phase and the sample solution, respectively, leading to the formation of a dynamic pH junction in the column and causing focus of alkaloids. Selective focus of analytes can be achieved on the basis of velocity changes of the pH junction through appropriate selection of solvent systems and optimization of additive concentrations. The extract can be directly introduced into the HSCCC for the online extraction, separation and purification of alkaloids from S. cepharantha. Continuous separation can be easily achieved with the same solvent system. Under the optimum conditions, 6.0 g original sample was extracted with 60 mL of the upper phase of hexane-ethyl acetate-methanol-water (1:1:1:1, v/v/v/v) containing 10% triethylamine under 50 °C and 400 W irradiation power for 10 min, the extracts were directly separated and purified by high-speed counter-current chromatography. A total of 5.7 mg sinomenine, 8.3mg 6,7-di-O-acetylsinococuline, 17.9 mg berbamine, 12.7 mg isotetrandrine and 14.6 mg cepharanthine were obtained with purities of 96.7%, 93.7%, 98.7%, 97.3% and 99.3%, respectively. The online method provides good selectivity to ionizable compounds and improves the separation and purification efficiency of the high-speed counter-current chromatography technique. It has good potential for separation and purification of effective compounds from natural products. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. Phyllomedusa bicolor skin secretion and the Kambô ritual

    PubMed Central

    2014-01-01

    The ritual of Kambô or Sapo is a type of voluntary envenomation. During this purification ritual a shaman healer, from various South American countries, deliberately burns the right shoulder with a glowing stick from a fireplace. Excretions of Phyllomedusa bicolor (or Giant Leaf Frog, Kambô or Sapo) are then applied to these fresh wounds. This ritual is used as a means of purification of the body, supposedly brings luck to hunters, increases stamina and enhances physical and sexual strength. All the peripheral and most of the central effects of the secretion can be ascribed to the exceptionally high content of active peptides, easily absorbed through burned skin. This article describes the ritual and the bio-active peptides from the secretion. PMID:26413084

  19. Phyllomedusa bicolor skin secretion and the Kambô ritual.

    PubMed

    den Brave, Paul S; Bruins, Eugéne; Bronkhorst, Maarten W G A

    2014-01-01

    The ritual of Kambô or Sapo is a type of voluntary envenomation. During this purification ritual a shaman healer, from various South American countries, deliberately burns the right shoulder with a glowing stick from a fireplace. Excretions of Phyllomedusa bicolor (or Giant Leaf Frog, Kambô or Sapo) are then applied to these fresh wounds. This ritual is used as a means of purification of the body, supposedly brings luck to hunters, increases stamina and enhances physical and sexual strength. All the peripheral and most of the central effects of the secretion can be ascribed to the exceptionally high content of active peptides, easily absorbed through burned skin. This article describes the ritual and the bio-active peptides from the secretion.

  20. Optimization of Growth Conditions for Purification and Production of L-Asparaginase by Spirulina maxima

    PubMed Central

    El Baroty, Gamal S.

    2016-01-01

    L-asparaginase (L-AsnA) is widely distributed among microorganisms and has important applications in medicine and in food technology sectors. Therefore, the ability of the production, purification, and characterization of AsnA from Spirulina maxima (SM) were tested. SM cultures grown in Zarrouk medium containing different N2 (in NaNO3 form) concentrations (1.25, 2.50, and 5.0 g/L) for 18 days contained a significant various quantity of dry biomass yields and AsnA enzyme levels. MS L-AsnA activity was found to be directly proportional to the N2 concentration. The cultures of SM at large scales (300 L medium, 5 g/L N2) showed a high AsnA enzyme activity (898 IU), total protein (405 mg/g), specific enzyme activity (2.21 IU/mg protein), and enzyme yield (51.28 IU/L) compared with those in low N2 cultures. The partial purification of crude MS AsnA enzyme achieved by 80% ammonium sulfate AS precipitated and CM-Sephadex C-200 gel filtration led to increases in the purification of enzyme with 5.28 and 10.91 times as great as that in SM crude enzymes. Optimum pH and temperature of purified AsnA for the hydrolyzate were 8.5 and 37 ± 0.2°C, respectively. To the best of our knowledge, this is the first report on L-asparaginase production in S. maxima. PMID:27525017

  1. A Fluorescent Tile DNA Diagnocode System for In Situ Rapid and Selective Diagnosis of Cytosolic RNA Cancer Markers

    PubMed Central

    Park, Kyung Soo; Shin, Seung Won; Jang, Min Su; Shin, Woojung; Yang, Kisuk; Min, Junhong; Cho, Seung-Woo; Oh, Byung-Keun; Bae, Jong Wook; Jung, Sunghwan; Choi, Jeong-Woo; Um, Soong Ho

    2015-01-01

    Accurate cancer diagnosis often requires extraction and purification of genetic materials from cells, and sophisticated instrumentations that follow. Otherwise in order to directly treat the diagnostic materials to cells, multiple steps to optimize dose concentration and treatment time are necessary due to diversity in cellular behaviors. These processes may offer high precision but hinder fast analysis of cancer, especially in clinical situations that need rapid detection and characterization of cancer. Here we present a novel fluorescent tile DNA nanostructure delivered to cancer cytosol by employing nanoparticle technology. Its structural anisotropicity offers easy manipulation for multifunctionalities, enabling the novel DNA nanostructure to detect intracellular cancer RNA markers with high specificity within 30 minutes post treatment, while the nanoparticle property bypasses the requirement of treatment optimization, effectively reducing the complexity of applying the system for cancer diagnosis. Altogether, the system offers a precise and rapid detection of cancer, suggesting the future use in the clinical fields. PMID:26678430

  2. Extracorporeal blood purification therapy for sepsis and systemic inflammation: its biological rationale.

    PubMed

    Bellomo, R; Baldwin, I; Ronco, C

    2001-01-01

    EBPTs represent a promising new approach to the adjuvant treatment of severe sepsis, septic shock and MODS. Their technology is rapidly evolving and pilot animal and human studies are now taking place to prepare the territory for the first large randomized controlled trial. The rationale for EBPT is reasonable and the initial data are encouraging. The correct technology and molecular targeting, however, are still being explored. Once the best technology has been determined, it is likely that phase II and phase III trials will be performed to test the hypothesis that these therapies can indeed alter mortality in severe inflammatory multiorgan dysfunction.

  3. The gas heterogeneous flows cleaning technology from corona discharge field

    NASA Astrophysics Data System (ADS)

    Bogdanov, A.; Tokarev, A.; Judanov, V.; Vinogradov, V.

    2017-11-01

    A nanogold capture and extraction from combustion products of Kara-Keche coal, description the process: a coal preparation to experiments, nanogold introducing in its composition, temperature and time performance of combustion, device and function of experimental apparatus, gas-purification of the gas flow process and receiving combustion products (condensate, coke, ash, rags) is offerred.

  4. Water Purification, Distribution and Sewage Disposal. Appropriate Technologies for Development. Reprint R-29.

    ERIC Educational Resources Information Center

    1979

    This document, designed to serve as a training manual for technical instructors and as a field resource reference for Peace Corps volunteers, consists of nine units. Unit topics focus on: (1) water supply sources; (2) water treatment; (3) planning water distribution systems; (4) characteristics of an adequate system; (5) construction techniques;…

  5. Advances in primary recovery: centrifugation and membrane technology.

    PubMed

    Roush, David J; Lu, Yuefeng

    2008-01-01

    Significant and continual improvements in upstream processing for biologics have resulted in challenges for downstream processing, both primary recovery and purification. Given the high cell densities achievable in both microbial and mammalian cell culture processes, primary recovery can be a significant bottleneck in both clinical and commercial manufacturing. The combination of increased product titer and low viability leads to significant relative increases in the levels of process impurities such as lipids, intracellular proteins and nucleic acid versus the product. In addition, cell culture media components such as soy and yeast hydrolysates have been widely applied to achieve the cell culture densities needed for higher titers. Many of the process impurities can be negatively charged at harvest pH and can form colloids during the cell culture and harvest processes. The wide size distribution of these particles and the potential for additional particles to be generated by shear forces within a centrifuge may result in insufficient clarification to prevent fouling of subsequent filters. The other residual process impurities can lead to precipitation and increased turbidity during processing and even interference with the performance of the capturing chromatographic step. Primary recovery also poses significant challenges owing to the necessity to execute in an expedient manner to minimize both product degradation and bioburden concerns. Both microfiltration and centrifugation coupled with depth filtration have been employed successfully as primary recovery processing steps. Advances in the design and application of membrane technology for microfiltration and dead-end filtration have contributed to significant improvements in process performance and integration, in some cases allowing for a combination of multiple unit operations in a given step. Although these advances have increased productivity and reliability, the net result is that optimization of primary recovery processes has become substantially more complicated. Ironically, the application of classical chemical engineering approaches to overcome issues in primary recovery and purification (e.g., turbidity and trace impurity removal) are just recently gaining attention. Some of these techniques (e.g., membrane cascades, pretreatment, precipitation, and the use of affinity tags) are now seen almost as disruptive technologies. This paper will review the current and potential future state of research on primary recovery, including relevant papers presented at the 234th American Chemical Society (ACS) National Meeting in Boston.

  6. Novel Self-driven Microbial Nutrient Recovery Cell with Simultaneous Wastewater Purification

    PubMed Central

    Chen, Xi; Sun, Dongya; Zhang, Xiaoyuan; Liang, Peng; Huang, Xia

    2015-01-01

    Conventional wastewater purification technologies consume large amounts of energy, while the abundant chemical energy and nutrient resources contained in sewage are wasted in such treatment processes. A microbial nutrient recovery cell (MNRC) has been developed to take advantage of the energy contained in wastewater, in order to simultaneously purify wastewater and recover nutrient ions. When wastewater was circulated between the anode and cathode chambers of the MNRC, the organics (COD) were removed by bacteria while ammonium and phosphate (NH4+-N and PO43−-P) were recovered by the electrical field that was produced using in situ energy in the wastewater without additional energy input. The removal efficiencies from wastewater were >82% for COD, >96% for NH4+-N, and >64% for PO43−-P in all the operational cycles. Simultaneously, the concentrations of NH4+ and PO43− in the recovery chamber increased to more than 1.5 and 2.2 times, respectively, compared with the initial concentrations in wastewater. The MNRC provides proof-of-concept as a sustainable, self-driven approach to efficient wastewater purification and nutrient recovery in a comprehensive bioelectrochemical system. PMID:26503712

  7. Purification and mass spectrometry based characterization of a pediocin produced by Pediococcus acidilactici 13.

    PubMed

    Altuntaş, Evrim Güneş; Ayhan, Kamuran; Peker, Selen; Ayhan, Beycan; Demiralp, Duygu Ozel

    2014-10-01

    Bacteriocins are antimicrobial peptides produced by several bacterial species. Among the bacteriocins pediocin-like bacteriocins have a significant inhibitory activity on the foodborne pathogens especially on Listeria monocytogenes. This study aims to select a simple and usable purification method to purify/concentrate the antimicrobial peptide and characterization of the bacteriocin produced by Pediococcus acidilactici 13 by using proteomic approaches which is a recent omic technology. For purification dialysis, ultrafiltration method was used, and as a result of this study the bacteriocin activity reached 819,200 AU/mL from 102,400 AU/mL initially. Two dimensional gel electrophoresis and then matrix-assisted laser desorption ionization/time of flight mass spectrometry (MALDI-TOF MS) analysis were carried out to identify the current bacteriocin and related proteins. Obtained data revealed similarity to pediocin PA-1 transport/processing ATP-binding protein PedD (accession number: P36497), pediocin operon PedC (accession number: Q68GC4) and bacteriocin pediocin PA-1 (accession number: P29430) from UniProtKB/Swiss-Prot databank, thus the bacteriocin produced by P. acidilactici 13 is considered similar to pediocin PA-1.

  8. Expression and purification of the matrix protein of Nipah virus in baculovirus insect cell system.

    PubMed

    Masoomi Dezfooli, Seyedehsara; Tan, Wen Siang; Tey, Beng Ti; Ooi, Chien Wei; Hussain, Siti Aslina

    2016-01-01

    Nipah virus (NiV) causes fatal respiratory illness and encephalitis in humans and animals. The matrix (M) protein of NiV plays an important role in the viral assembly and budding process. Thus, an access to the NiV M protein is vital to the design of viral antigens as diagnostic reagents. In this study, recombinant DNA technology was successfully adopted in the cloning and expression of NiV M protein. A recombinant expression cassette (baculovirus expression vector) was used to encode an N-terminally His-tagged NiV M protein in insect cells. A time-course study demonstrated that the highest yield of recombinant M protein (400-500 μg) was expressed from 107 infected cells 3 days after infection. A single-step purification method based on metal ion affinity chromatography was established to purify the NiV M protein, which successfully yielded a purity level of 95.67% and a purification factor of 3.39. The Western blotting and enzyme-linked immunosorbent assay (ELISA) showed that the purified recombinant M protein (48 kDa) was antigenic and reacted strongly with the serum of a NiV infected pig. © 2015 American Institute of Chemical Engineers.

  9. M13 bacteriophage purification using poly(ionic liquids) as alternative separation matrices.

    PubMed

    Jacinto, Maria João; Patinha, David J S; Marrucho, Isabel M; Gonçalves, João; Willson, Richard C; Azevedo, Ana M; Aires-Barros, M Raquel

    2018-01-12

    M13 is a filamentous, non-lytic bacteriophage that infects Escherichia coli via the F pilus. Currently, phage M13 is widely used in phage display technology and bio-nanotechnology, and is considered a possible antibacterial therapeutic agent, among other applications. Conventional phage purification involves 5-7 operational steps, with high operational costs and significant product loss (approximately 60%). In this work, we propose a scalable purification process for M13 bacteriophage using a novel stationary phase based on a polymeric ionic liquid (PIL) with a positively charged backbone structure. Poly (1-vinyl-3-ethyl imidazolium bis(trifluoromethylsulfonyl) imide) - poly(VEIM-TFSI) predominantly acted as an anion exchanger under binding-elution mode. This revealed to be a rapid and simple method for the recovery of phage M13 with an overall separation yield of over 70% after a single downstream step. To the best of our knowledge, PILs have never been used as separation matrices for biological products and the results obtained, together with the large number of cations and anions available to prepare PILs, illustrate well the large potential of the proposed methodology. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Purification of bacteriophage M13 by anion exchange chromatography.

    PubMed

    Monjezi, Razieh; Tey, Beng Ti; Sieo, Chin Chin; Tan, Wen Siang

    2010-07-01

    M13 is a non-lytic filamentous bacteriophage (phage). It has been used widely in phage display technology for displaying foreign peptides, and also for studying macromolecule structures and interactions. Traditionally, this phage has been purified by cesium chloride (CsCl) density gradient ultracentrifugation which is highly laborious and time consuming. In the present study, a simple, rapid and efficient method for the purification of M13 based on anion exchange chromatography was established. A pre-packed SepFast Super Q column connected to a fast protein liquid chromatography (FPLC) system was employed to capture released phages in clarified Escherichia coli fermented broth. An average yield of 74% was obtained from a packed bed mode elution using citrate buffer (pH 4), containing 1.5 M NaCl at 1 ml/min flow rate. The purification process was shortened substantially to less than 2 h from 18 h in the conventional ultracentrifugation method. SDS-PAGE revealed that the purity of particles was comparable to that of CsCl gradient density ultracentrifugation method. Plaque forming assay showed that the purified phages were still infectious. Copyright 2010 Elsevier B.V. All rights reserved.

  11. Magnetic Purification of Antibodies

    NASA Astrophysics Data System (ADS)

    Dhadge, Vijaykumar Laxman

    This work aimed at the development of magnetic nanoparticles for antibody purification and at the evaluation of their performance in Magnetic fishing and in a newly developed hybrid technology Magnetic Aqueous Two Phase Systems. Magnetic materials were produced by coprecipitation and solvothermal approaches. Natural polymers such as dextran, extracellular polysaccharide and gum Arabic were employed for coating of iron oxide magnetic supports. Polymer coated magnetic supports were then modified with synthetic antibody specific ligands,namely boronic acid, a triazine ligand (named 22/8) and an Ugi ligand (named A2C7I1). To optimize the efficacy of magnetic nanoparticles for antibody magnetic fishing, various solutions of pure and crude antibody solutions along with BSA as a non-specific binding protein were tested. The selectivity of magnetic nanoparticle for antibody, IgG, was found effective with boronic acid and ligand 22/8. Magnetic supports were then studied for their performance in high gradient magnetic separator for effective separation capability as well as higher volume handling capability. The magnetic materials were also supplemented to aqueous two phase systems, devising a new purification technology. For this purpose, magnetic particles modified with boronic acid were more effective. This alternative strategy reduced the time of operation,maximized separation capability (yield and purity), while reducing the amount of salt required. Boronic acid coated magnetic particles bound 170 +/- 10 mg hIgG/g MP and eluted 160 +/- 5 mg hIgG/g MP, while binding only 15 +/- 5 mg BSA/g MP. The affinity constant for the interaction between hIgG and APBA_MP was estimated as 4.9 x 105 M-1 (Ka) with a theoretical maximum capacity of 492 mg hIgG adsorbed/g MP (Qmax). APBA_MPs were also tested for antibody purification directly from CHO cell supernatants. The particles were able to bind 98% of IgG loaded and to recover 95% of pure IgG (purity greater than 98%) at extremely mild conditions. None None None None None None None None None None None None None None None None None None None None None None None None

  12. Technology of surface wastewater purification, including high-rise construction areas

    NASA Astrophysics Data System (ADS)

    Tsyba, Anna; Skolubovich, Yury

    2018-03-01

    Despite on the improvements in the quality of high-rise construction areas and industrial wastewater treatment, the pollution of water bodies continues to increase. This is due to the organized and unorganized surface untreated sewage entry into the reservoirs. The qualitative analysis of some cities' surface sewage composition is carried out in the work. Based on the published literature review, the characteristic contamination present in surface wastewater was identified. The paper proposes a new technology for the treatment of surface sewage and presents the results of preliminary studies.

  13. Recovery of Anthocyanins Using Membrane Technologies: A Review.

    PubMed

    Martín, Julia; Díaz-Montaña, Enrique Jacobo; Asuero, Agustin G

    2018-05-04

    Anthocyanins are naturally occurring polyphenolic compounds and give many flowers, fruits and vegetable their orange, red, purple and blue colors. Besides their color attributes, anthocyanins have received much attention in recent years due to the growing evidence of their antioxidant capacity and health benefits on humans. However, these compounds usually occur in low concentrations in mixtures of complex matrices, and therefore large-scale harvesting is needed to obtain sufficient amounts for their practical usage. Effective fractionation or separation technologies are therefore essential for the screening and production of these bioactive compounds. In this context, membrane technologies have become popular due to their operational simplicity, the capacity to achieve good simultaneous separation/pre-concentration and matrix reduction with lower temperature and lower operating cost in comparison to other sample preparation methods. Membrane fractionation is based on the molecular or particle sizes (pressure-driven processes), on their charge (electrically driven processes) or are dependent on both size and charge. Other non-pressure-driven membrane processes (osmotic pressure and vapor pressure-driven) have been developed in recent years and employed as alternatives for the separation or fractionation of bioactive compounds at ambient conditions without product deterioration. These technologies are applied either individually or in combination as an integrated membrane system to meet the different requirements for the separation of bioactive compounds. The first section of this review examines the basic principles of membrane processes, including the different types of membranes, their structure, morphology and geometry. The most frequently used techniques are also discussed. Last, the specific application of these technologies for the separation, purification and concentration of phenolic compounds, with special emphasis on anthocyanins, are also provided.

  14. Development of a Novel Self-Enclosed Sample Preparation Device for DNA/RNA Isolation in Space

    NASA Technical Reports Server (NTRS)

    Zhang, Ye; Mehta, Satish K.; Pensinger, Stuart J.; Pickering, Karen D.

    2011-01-01

    Modern biology techniques present potentials for a wide range of molecular, cellular, and biochemistry applications in space, including detection of infectious pathogens and environmental contaminations, monitoring of drug-resistant microbial and dangerous mutations, identification of new phenotypes of microbial and new life species. However, one of the major technological blockades in enabling these technologies in space is a lack of devices for sample preparation in the space environment. To overcome such an obstacle, we constructed a prototype of a DNA/RNA isolation device based on our novel designs documented in the NASA New Technology Reporting System (MSC-24811-1/3-1). This device is self-enclosed and pipette free, purposely designed for use in the absence of gravity. Our design can also be modified easily for preparing samples in space for other applications, such as flowcytometry, immunostaining, cell separation, sample purification and separation according to its size and charges, sample chemical labeling, and sample purification. The prototype of our DNA/RNA isolation device was tested for efficiencies of DNA and RNA isolation from various cell types for PCR analysis. The purity and integrity of purified DNA and RNA were determined as well. Results showed that our developed DNA/RNA isolation device offers similar efficiency and quality in comparison to the samples prepared using the standard protocol in the laboratory.

  15. Synthesis and Characterization of Copper Complexes with a Tridentate Nitrogen-Donor Ligand: An Integrated Research Experiment for Undergraduate Students

    ERIC Educational Resources Information Center

    Bussey, Katherine A.; Cavalier, Annie R.; Connell, Jennifer R.; Mraz, Margaret E.; Holderread, Ashley S.; Oshin, Kayode D.; Pintauer, Tomislav

    2015-01-01

    An integrated laboratory experiment applying concepts and techniques developed in organic chemistry, inorganic chemistry, and instrumental analysis is presented for use by students interested in undergraduate research. The experiment incorporates some advanced laboratory practices such as multistep organic synthesis and purification, detailed…

  16. Electromigration process for the purification of molten silicon during crystal growth

    DOEpatents

    Lovelace, Alan M. Administrator of the National Aeronautics and Space; Shlichta, Paul J.

    1982-01-01

    A process for the purification of molten materials during crystal growth by electromigration of impurities to localized dirty zones. The process has particular applications for silicon crystal growth according to Czochralski techniques and edge-defined film-fed growth (EFG) conditions. In the Czochralski crystal growing process, the impurities are electromigrated away from the crystallization interface by applying a direct electrical current to the molten silicon for electromigrating the charged impurities away from the crystal growth interface. In the EFG crystal growth process, a direct electrical current is applied between the two faces which are used in forming the molten silicon into a ribbon. The impurities are thereby migrated to one side only of the crystal ribbon. The impurities may be removed or left in place. If left in place, they will not adversely affect the ribbon when used in solar collectors. The migration of the impurity to one side only of the silicon ribbon is especially suitable for use with asymmetric dies which preferentially crystallize uncharged impurities along one side or face of the ribbon.

  17. Microbial-assisted synthesis and evaluation the cytotoxic effect of tellurium nanorods.

    PubMed

    Forootanfar, Hamid; Amirpour-Rostami, Sahar; Jafari, Mandana; Forootanfar, Amir; Yousefizadeh, Zahra; Shakibaie, Mojtaba

    2015-04-01

    The present study was designed to isolate bacterial strain capable of tellurium nanorods' (Te NRs) production followed by purification and evaluation of the cytotoxic effect of Te NRs. Among 25 environmental samples collected for screening of Te NR-producer bacterial strains one bacterial colony (isolated from hot spring and identified as Pseudomonas pseudoalcaligenes strain Te) was selected and applied for biosynthesis of Te NRs. Thereafter, an organic-aqueous partitioning system was applied for the purification of the biogenic Te NRs and the purified Te NRs were characterized using transmission electron microscopy (TEM), scanning electron microscopy (SEM), energy dispersive X-ray (EDX), X-ray diffraction spectroscopy (XRD), UV-visible spectroscopy, and Fourier transform infrared spectroscopy (FTIR) techniques. The cytotoxic effect of biologically synthesized Te NRs and potassium tellurite on four cell lines of MCF-7, HT1080, HepG2 and A549 was then determined using the MTT assay method. The obtained results revealed lower toxicity for the rod-shaped biogenic tellurium nanostructures (~22nm diameter by 185nm length) compared to K2TeO3. Copyright © 2014. Published by Elsevier B.V.

  18. Optimization of elution salt concentration in stepwise elution of protein chromatography using linear gradient elution data. Reducing residual protein A by cation-exchange chromatography in monoclonal antibody purification.

    PubMed

    Ishihara, Takashi; Kadoya, Toshihiko; Endo, Naomi; Yamamoto, Shuichi

    2006-05-05

    Our simple method for optimization of the elution salt concentration in stepwise elution was applied to the actual protein separation system, which involves several difficulties such as detection of the target. As a model separation system, reducing residual protein A by cation-exchange chromatography in human monoclonal antibody (hMab) purification was chosen. We carried out linear gradient elution experiments and obtained the data for the peak salt concentration of hMab and residual protein A, respectively. An enzyme-linked immunosorbent assay was applied to the measurement of the residual protein A. From these data, we calculated the distribution coefficient of the hMab and the residual protein A as a function of salt concentration. The optimal salt concentration of stepwise elution to reduce the residual protein A from the hMab was determined based on the relationship between the distribution coefficient and the salt concentration. Using the optimized condition, we successfully performed the separation, resulting in high recovery of hMab and the elimination of residual protein A.

  19. Feasibility study of NaOH regeneration in acid gas removal unit using membrane electrolysis

    NASA Astrophysics Data System (ADS)

    Taufany, Fadlilatul; Pratama, Alvian; Romzuddin, Muhammad

    2017-05-01

    The world's energy demand is increasing with the development of human civilization. Due to limited energy resource, after 2020 fossil fuels thus is predicted will be replaced by renewable resources. Taking an example, one of the potential renewable energy to be considered is biogas, as its high content of methane, which can be produced via the fermentation process of the organic compounds under controlled anaerobic environment by utilizing the methanogen bacteria. However, prior the further use, this biogas must be purified from its impurities contents, i.e. acid gas of CO2 and H2S, up to 4% and 16 ppmv, respectively, in the acid gas removal unit. This such of purification efforts, will significantly increase the higher heating value of biogas, approximately from 600 to 900 Btu/Scf. During the purification process in this acid gas removal unit, NaOH solution is used as a liquid absorbent to reduce those acid gases content, in which the by-product of alkali salt (brine) was produced as waste. Here we report the feasibility study of the NaOH regeneration process in acid gas removal unit via membrane electrolysis technology, in which both the technical and economic aspects are taken account. To be precise in procedure, the anode semi-cell was filled with the brine solution, while the cathode semi-cell was filled with demineralized water, and those electrodes were separated by the cation exchange membrane. Furthermore, the applied potential was varied ranging from 5, 10, 15 and to 20 V, while the concentration of KCl electrolyte solutions were varied ranging from 0.01, 0.05, 0.1, and to 0.03 M. This study was conducted under controlled temperatures of 30 and 50 °C. Here we found that the % sodium recovery was increased along with the applied potential, temperature, and the decrease in KCl electrolyte concentration. We found that the best results, by means of the highest % sodium recovery, i.e. 97.26 %, was achieved under the experimental condition of temperature at 30 °C, applied potential at 15 V, and KCl electrolyte concentration at 0.01 M. At such electrolysis condition, the energy efficiency was calculated to be 0,009 M-NaOH/Wh, or was equal to operating cost at 0.04/kg-NaOH.

  20. High-performance liquid chromatography of human glycoprotein hormones.

    PubMed

    Chlenov, M A; Kandyba, E I; Nagornaya, L V; Orlova, I L; Volgin, Y V

    1993-02-12

    The chromatographic behavior of the glycoprotein hormones from human pituitary glands and of placental origin [thyroid-stimulating hormone, luteinizing hormone and chorionic gonadotropin (CG)] was studied. It was shown that hydrophobic interaction chromatography on a microparticulate packing and anion-exchange HPLC can be applied for the purification of these hormones. Reversed-phase HPLC on wide-pore C4-bonded silica at neutral pH can be applied for the determination of the above hormones and for the isolation of pure CG and its subunits.

  1. Liquid membrane purification of biogas

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Majumdar, S.; Guha, A.K.; Lee, Y.T.

    1991-03-01

    Conventional gas purification technologies are highly energy intensive. They are not suitable for economic removal of CO{sub 2} from methane obtained in biogas due to the small scale of gas production. Membrane separation techniques on the other hand are ideally suited for low gas production rate applications due to their modular nature. Although liquid membranes possess a high species permeability and selectivity, they have not been used for industrial applications due to the problems of membrane stability, membrane flooding and poor operational flexibility, etc. A new hollow-fiber-contained liquid membrane (HFCLM) technique has been developed recently. This technique overcomes the shortcomingsmore » of the traditional immobilized liquid membrane technology. A new technique uses two sets of hydrophobic, microporous hollow fine fibers, packed tightly in a permeator shell. The inter-fiber space is filled with an aqueous liquid acting as the membrane. The feed gas mixture is separated by selective permeation of a species through the liquid from one fiber set to the other. The second fiber set carries a sweep stream, gas or liquid, or simply the permeated gas stream. The objectives (which were met) of the present investigation were as follows. To study the selective removal of CO{sub 2} from a model biogas mixture containing 40% CO{sub 2} (the rest being N{sub 2} or CH{sub 4}) using a HFCLM permeator under various operating modes that include sweep gas, sweep liquid, vacuum and conventional permeation; to develop a mathematical model for each mode of operation; to build a large-scale purification loop and large-scale permeators for model biogas separation and to show stable performance over a period of one month.« less

  2. Towards Plasma-Based Water Purification: Challenges and Prospects for the Future

    NASA Astrophysics Data System (ADS)

    Foster, John

    2016-10-01

    Freshwater scarcity derived from climate change, pollution, and over-development has led to serious consideration for water reuse. Advanced water treatment technologies will be required to process wastewater slated for reuse. One new and emerging technology that could potentially address the removal micropollutants in both drinking water as well as wastewater slated for reuse is plasma-based water purification. Plasma in contact with liquid water generates reactive species that attack and ultimately mineralize organic contaminants in solution. This interaction takes place in a boundary layer centered at the plasma-liquid interface. An understanding of the physical processes taking place at this interface, though poorly understood, is key to the optimization of plasma water purifiers. High electric field conditions, large density gradients, plasma-driven chemistries, and fluid dynamic effects prevail in this multiphase region. The region is also the source function for longer-lived reactive species that ultimately treat the water. Here, we review the need for advanced water treatment methods and in the process, make the case for plasma-based methods. Additionally, we survey the basic methods of interacting plasma with liquid water (including a discussion of breakdown processes in water), the current state of understanding of the physical processes taking place at the plasma-liquid interface, and the role that these processes play in water purification. The development of diagnostics usable in this multiphase environment along modeling efforts aimed at elucidating physical processes taking place at the interface are also detailed. Key experiments that demonstrate the capability of plasma-based water treatment are also reviewed. The technical challenges to the implementation of plasma-based water reactors are also discussed. NSF CBET 1336375 and DOE DE-SC0001939.

  3. Coagulant recovery and reuse for drinking water treatment.

    PubMed

    Keeley, James; Jarvis, Peter; Smith, Andrea D; Judd, Simon J

    2016-01-01

    Coagulant recovery and reuse from waterworks sludge has the potential to significantly reduce waste disposal and chemicals usage for water treatment. Drinking water regulations demand purification of recovered coagulant before they can be safely reused, due to the risk of disinfection by-product precursors being recovered from waterworks sludge alongside coagulant metals. While several full-scale separation technologies have proven effective for coagulant purification, none have matched virgin coagulant treatment performance. This study examines the individual and successive separation performance of several novel and existing ferric coagulant recovery purification technologies to attain virgin coagulant purity levels. The new suggested approach of alkali extraction of dissolved organic compounds (DOC) from waterworks sludge prior to acidic solubilisation of ferric coagulants provided the same 14:1 selectivity ratio (874 mg/L Fe vs. 61 mg/L DOC) to the more established size separation using ultrafiltration (1285 mg/L Fe vs. 91 mg/L DOC). Cation exchange Donnan membranes were also examined: while highly selective (2555 mg/L Fe vs. 29 mg/L DOC, 88:1 selectivity), the low pH of the recovered ferric solution impaired subsequent treatment performance. The application of powdered activated carbon (PAC) to ultrafiltration or alkali pre-treated sludge, dosed at 80 mg/mg DOC, reduced recovered ferric DOC contamination to <1 mg/L but in practice, this option would incur significant costs. The treatment performance of the purified recovered coagulants was compared to that of virgin reagent with reference to key water quality parameters. Several PAC-polished recovered coagulants provided the same or improved DOC and turbidity removal as virgin coagulant, as well as demonstrating the potential to reduce disinfection byproducts and regulated metals to levels comparable to that attained from virgin material. Copyright © 2015 Elsevier Ltd. All rights reserved.

  4. Fusion tags for protein solubility, purification and immunogenicity in Escherichia coli: the novel Fh8 system

    PubMed Central

    Costa, Sofia; Almeida, André; Castro, António; Domingues, Lucília

    2014-01-01

    Proteins are now widely produced in diverse microbial cell factories. The Escherichia coli is still the dominant host for recombinant protein production but, as a bacterial cell, it also has its issues: the aggregation of foreign proteins into insoluble inclusion bodies is perhaps the main limiting factor of the E. coli expression system. Conversely, E. coli benefits of cost, ease of use and scale make it essential to design new approaches directed for improved recombinant protein production in this host cell. With the aid of genetic and protein engineering novel tailored-made strategies can be designed to suit user or process requirements. Gene fusion technology has been widely used for the improvement of soluble protein production and/or purification in E. coli, and for increasing peptide’s immunogenicity as well. New fusion partners are constantly emerging and complementing the traditional solutions, as for instance, the Fh8 fusion tag that has been recently studied and ranked among the best solubility enhancer partners. In this review, we provide an overview of current strategies to improve recombinant protein production in E. coli, including the key factors for successful protein production, highlighting soluble protein production, and a comprehensive summary of the latest available and traditionally used gene fusion technologies. A special emphasis is given to the recently discovered Fh8 fusion system that can be used for soluble protein production, purification, and immunogenicity in E. coli. The number of existing fusion tags will probably increase in the next few years, and efforts should be taken to better understand how fusion tags act in E. coli. This knowledge will undoubtedly drive the development of new tailored-made tools for protein production in this bacterial system. PMID:24600443

  5. Integration of gas chromatography mass spectrometry methods for differentiating ricin preparation methods.

    PubMed

    Wunschel, David S; Melville, Angela M; Ehrhardt, Christopher J; Colburn, Heather A; Victry, Kristin D; Antolick, Kathryn C; Wahl, Jon H; Wahl, Karen L

    2012-05-07

    The investigation of crimes involving chemical or biological agents is infrequent, but presents unique analytical challenges. The protein toxin ricin is encountered more frequently than other agents and is found in the seeds of Ricinus communis, commonly known as the castor plant. Typically, the toxin is extracted from castor seeds utilizing a variety of different recipes that result in varying purity of the toxin. Moreover, these various purification steps can also leave or differentially remove a variety of exogenous and endogenous residual components with the toxin that may indicate the type and number of purification steps involved. We have applied three gas chromatography-mass spectrometry (GC-MS) based analytical methods to measure the variation in seed carbohydrates and castor oil ricinoleic acid, as well as the presence of solvents used for purification. These methods were applied to the same samples prepared using four previously identified toxin preparation methods, starting from four varieties of castor seeds. The individual data sets for seed carbohydrate profiles, ricinoleic acid, or acetone amount each provided information capable of differentiating different types of toxin preparations across seed types. However, the integration of the data sets using multivariate factor analysis provided a clear distinction of all samples based on the preparation method, independent of the seed source. In particular, the abundance of mannose, arabinose, fucose, ricinoleic acid, and acetone were shown to be important differentiating factors. These complementary tools provide a more confident determination of the method of toxin preparation than would be possible using a single analytical method.

  6. Advanced Continuous Flow Platform for On-Demand Pharmaceutical Manufacturing.

    PubMed

    Zhang, Ping; Weeranoppanant, Nopphon; Thomas, Dale A; Tahara, Kohei; Stelzer, Torsten; Russell, Mary Grace; O'Mahony, Marcus; Myerson, Allan S; Lin, Hongkun; Kelly, Liam P; Jensen, Klavs F; Jamison, Timothy F; Dai, Chunhui; Cui, Yuqing; Briggs, Naomi; Beingessner, Rachel L; Adamo, Andrea

    2018-02-21

    As a demonstration of an alternative to the challenges faced with batch pharmaceutical manufacturing including the large production footprint and lengthy time-scale, we previously reported a refrigerator-sized continuous flow system for the on-demand production of essential medicines. Building on this technology, herein we report a second-generation, reconfigurable and 25 % smaller (by volume) continuous flow pharmaceutical manufacturing platform featuring advances in reaction and purification equipment. Consisting of two compact [0.7 (L)×0.5 (D)×1.3 m (H)] stand-alone units for synthesis and purification/formulation processes, the capabilities of this automated system are demonstrated with the synthesis of nicardipine hydrochloride and the production of concentrated liquid doses of ciprofloxacin hydrochloride, neostigmine methylsulfate and rufinamide that meet US Pharmacopeia standards. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Air Revitalization System Enables Excursions to the Stratosphere

    NASA Technical Reports Server (NTRS)

    2015-01-01

    Paragon Space Development Corporation, based in Tucson, Arizona has had a long history of collaboration with NASA, including developing a modular air purification system under the Commercial Crew Development Program, designed to support the commercial space sector. Using that device and other NASA technology, startup company World View is now gearing up to take customers on helium balloon rides to the stratosphere.

  8. Charged Particles Kill Pathogens and Round Up Dust

    NASA Technical Reports Server (NTRS)

    2015-01-01

    To keep plants fresh longer in space, Marshall Space Flight Center awarded funding to the University of Wisconsin-Madison to develop a titanium oxide-based device that reduced the amount of decay-inducing ethylene gas in the air. Electrolux (now Dallas-based Aerus Holdings) furthered the technology by developing an air purification product that kills pathogens both in the atmosphere and on surfaces.

  9. JPRS Report, Environmental Issues

    DTIC Science & Technology

    1990-06-13

    in settlements in valley locations, with the simultaneous application of efficient waste- gas purification proce- dures (dedusting, desulfurization ...Slaughtered [RUDE PRAVO 1 JunJ 17 Power Plant Desulfurization Technology, Funding Explored [ZEMEDELSKE NOVINY 29 Apr] . 17 GERMAN DEMOCRATIC...the Soviet Union, it is a question of money and a shortage of foreign currency but apparently also an attempt to develop its own pollution control

  10. JPRS Report, Science & Technology, Japan

    DTIC Science & Technology

    1988-11-10

    stage of microbial culture and fermentation through the stage of product isolation and purification. The information is expected to become an...controlled so that a process can be manipulated normally and safely. For example, in a bioreactor process mainly involving fermentation , the author and his...10) Electron microscope (11) Jar fermenter Continuous centrifuge Examples of equipment used in cell fusion research (1) Autotable Heated vibrating

  11. Electrodrift purification of materials for room temperature radiation detectors

    DOEpatents

    James, R.B.; Van Scyoc, J.M. III; Schlesinger, T.E.

    1997-06-24

    A method of purifying nonmetallic, crystalline semiconducting materials useful for room temperature radiation detecting devices by applying an electric field across the material is disclosed. The present invention discloses a simple technology for producing purified ionic semiconducting materials, in particular PbI{sub 2} and preferably HgI{sub 2}, which produces high yields of purified product, requires minimal handling of the material thereby reducing the possibility of introducing or reintroducing impurities into the material, is easy to control, is highly selective for impurities, retains the stoichiometry of the material and employs neither high temperatures nor hazardous materials such as solvents or liquid metals. An electric field is applied to a bulk sample of the material causing impurities present in the sample to drift in a preferred direction. After all of the impurities have been transported to the ends of the sample the current flowing through the sample, a measure of the rate of transport of mobile impurities, falls to a low, steady state value, at which time the end sections of the sample where the impurities have concentrated are removed leaving a bulk sample of higher purity material. Because the method disclosed here only acts on the electrically active impurities, the stoichiometry of the host material remains substantially unaffected. 4 figs.

  12. Electrodrift purification of materials for room temperature radiation detectors

    DOEpatents

    James, Ralph B.; Van Scyoc, III, John M.; Schlesinger, Tuviah E.

    1997-06-24

    A method of purifying nonmetallic, crystalline semiconducting materials useful for room temperature radiation detecting devices by applying an electric field across the material. The present invention discloses a simple technology for producing purified ionic semiconducting materials, in particular PbI.sub.2 and preferably HgI.sub.2, which produces high yields of purified product, requires minimal handling of the material thereby reducing the possibility of introducing or reintroducing impurities into the material, is easy to control, is highly selective for impurities, retains the stoichiometry of the material and employs neither high temperatures nor hazardous materials such as solvents or liquid metals. An electric field is applied to a bulk sample of the material causing impurities present in the sample to drift in a preferred direction. After all of the impurities have been transported to the ends of the sample the current flowing through the sample, a measure of the rate of transport of mobile impurities, falls to a low, steady state value, at which time the end sections of the sample where the impurities have concentrated are removed leaving a bulk sample of higher purity material. Because the method disclosed here only acts on the electrically active impurities, the stoichiometry of the host material remains substantially unaffected.

  13. Novel Nanotube Manufacturing Streamlines Production

    NASA Technical Reports Server (NTRS)

    2007-01-01

    Nanotubes have novel qualities that make them uniquely qualified for a plethora of uses, including applications in electronics, optics, and other scientific and industrial fields. The NASA process for creating these nanostructures involves using helium arc welding to vaporize an amorphous carbon rod and then form nanotubes by depositing the vapor onto a water-cooled carbon cathode, which then yields bundles, or ropes, of single-walled nanotubes at a rate of 2 grams per hour using a single setup. This eliminates costs associated with the use of metal catalysts, including the cost of product purification, resulting in a relatively inexpensive, high-quality, very pure end product. While managing to be less expensive, safer, and simpler, the process also increases the quality of the nanotubes. Goddard's Innovative Partnerships Program (IPP) Office promoted the technology, and in 2005, Boise-based Idaho Space Materials Inc. (ISM) was formed and applied for a nonexclusive license for the single-walled carbon nanotube (SWCNT) manufacturing technology. ISM commercialized its products, and the inexpensive, robust nanotubes are now in the hands of the scientists who will create the next generation of composite polymers, metals, and ceramics that will impact the way we live. In fact, researchers are examining ways for these newfound materials to be used in the manufacture of transistors and fuel cells, large screen televisions, ultra-sensitive sensors, high-resolution atomic force microscopy probes, supercapacitors, transparent conducting films, drug carriers, catalysts, and advanced composite materials, to name just a few of the myriad technologies to benefit.

  14. Preparation of convection interaction media isobutyl disc monolithic column and its application to purification of secondary alcohol dehydrogenase and alcohol oxidase.

    PubMed

    Isobe, Kimiyasu; Kawakami, Yoshimitsu

    2007-03-09

    A convection interaction media (trade name CIM, BIA Separation, Ljubljana, Slovenia) isobutyl monolithic disc was prepared by incubating a CIM epoxy monolithic disc with isobutylamine, and it was then applied to the purification of secondary alcohol dehydrogenase (S-ADH) and primary alcohol oxidase (P-AOD). Both enzymes were adsorbed on this column and eluted with high purity. Thus, S-ADH was purified to an electrophoretically homogeneous state by four column chromatographies using CIM DEAE-8 and CIM C4-8 tube monolithic columns, blue-Sepharose column and CIM isobutyl disc monolithic column. P-AOD was also purified to an electrophoretically homogeneous state by three column chromatographies of CIM DEAE-8 tube, CIM C4-8 tube and CIM isobutyl disc columns.

  15. Purification of angiotensin I-converting enzyme (ACE) inhibitory peptides from casein hydrolysate by IMAC-Ni2.

    PubMed

    Wu, Shanguang; Feng, Xuezhen; Lu, Yuan; Lu, Yuting; Liu, Saisai; Tian, Yuhong

    2017-10-01

    Casein proteins were hydrolyzed by papain to identify inhibitory peptides of angiotensin I-converting enzyme (ACE). The hydrolysate was fractionized by immobilized metal affinity chromatography (IMAC-Ni 2+ ). The fraction with high ACE inhibitory activity was enriched and further chromatographed on a reverse-phase column to yield four fractions. Among the fractions, the L4 fraction exhibited the highest ACE inhibitory activity and was identified by sequence analysis as Trp-Tyr-Leu-His-Tyr-Ala (WYLHYA), with IC 50 value of 16.22 ± 0.83 µM in vitro. This peptide was expected to be applied as an ingredient for preventing hypertension and IMAC-Ni 2+ may provide a simple method for purification of ACE inhibitory peptides.

  16. Purification of nattokinase by reverse micelles extraction from fermentation broth: effect of temperature and phase volume ratio.

    PubMed

    Liu, Jun-Guo; Xing, Jian-Min; Chang, Tian-Shi; Liu, Hui-Zhou

    2006-03-01

    Nattokinase is a novel fibrinolytic enzyme that is considered to be a promising agent for thrombosis therapy. In this study, reverse micelles extraction was applied to purify and concentrate nattokinase from fermentation broth. The effects of temperature and phase volume ratio used for the forward and backward extraction on the extraction process were examined. The optimal temperature for forward and backward extraction were 25 degrees C and 35 degrees C respectively. Nattokinase became more thermosensitive during reverse micelles extraction. And it could be enriched in the stripping phase eight times during backward extraction. It was found that nattokinase could be purified by AOT reverse micelles with up to 80% activity recovery and with a purification factor of 3.9.

  17. Radiation processing applications in the Czechoslovak water treatment technologies

    NASA Astrophysics Data System (ADS)

    Vacek, K.; Pastuszek, F.; Sedláček, M.

    The regeneration of biologically clogged water wells by radiation proved to be a successful and economically beneficial process among other promising applications of ionizing radiation in the water supply technology. The application conditions and experience are mentioned. The potential pathogenic Mycobacteria occuring in the warm washing and bathing water are resistant against usual chlorine and ozone concentrations. The radiation sensitivity of Mycobacteria allowed to suggest a device for their destroying by radiation. Some toxic substances in the underground water can be efficiently degraded by gamma radiation directly in the wells drilled as a hydraulic barrier surrounding the contaminated land area. Substantial decrease of CN - concentration and C.O.D. value was observed in water pumped from such well equipped with cobalt sources and charcoal. The removing of pathogenic contamination remains to be the main goal of radiation processing in the water purification technologies. The decrease of liquid sludge specific filter resistance and sedimentation acceleration by irradiation have a minor technological importance. The hygienization of sludge cake from the mechanical belt filter press by electron beam appears to be the optimum application in the Czechoslovak conditions. The potatoes and barley crop yields from experimental plots treated with sludge were higher in comparison with using the manure. Biological sludge from the municipal and food industry water purification plants contains nutritive components. The proper hygienization is a necessary condition for using them as a livestock feed supplement. Feeding experiments with broilers and pigs confirmed the possibility of partial (e.g. 50%) replacement of soya-, bone- or fish flour in feed mixtures by dried sludge hygienized either by heat or by the irradiation.

  18. Electron beam technology for multipollutant emissions control from heavy fuel oil-fired boiler.

    PubMed

    Chmielewski, Andrzej G; Ostapczuk, Anna; Licki, Janusz

    2010-08-01

    The electron beam treatment technology for purification of exhaust gases from the burning of heavy fuel oil (HFO) mazout with sulfur content approximately 3 wt % was tested at the Institute of Nuclear Chemistry and Technology laboratory plant. The parametric study was conducted to determine the sulfur dioxide (SO2), oxides of nitrogen (NO(x)), and polycyclic aromatic hydrocarbon (PAH) removal efficiency as a function of temperature and humidity of irradiated gases, absorbed irradiation dose, and ammonia stoichiometry process parameters. In the test performed under optimal conditions with an irradiation dose of 12.4 kGy, simultaneous removal efficiencies of approximately 98% for SO2, and 80% for NO(x) were recorded. The simultaneous decrease of PAH and one-ringed aromatic hydrocarbon (benzene, toluene, and xylenes [BTX]) concentrations was observed in the irradiated flue gas. Overall removal efficiencies of approximately 42% for PAHs and 86% for BTXs were achieved with an irradiation dose 5.3 kGy. The decomposition ratio of these compounds increased with an increase of absorbed dose. The decrease of PAH and BTX concentrations was followed by the increase of oxygen-containing aromatic hydrocarbon concentrations. The PAH and BTX decomposition process was initialized through the reaction with hydroxyl radicals that formed in the electron beam irradiated flue gas. Their decomposition process is based on similar principles as the primary reaction concerning SO2 and NO(x) removal; that is, free radicals attack organic compound chains or rings, causing volatile organic compound decomposition. Thus, the electron beam flue gas treatment (EBFGT) technology ensures simultaneous removal of acid (SO2 and NO(x)) and organic (PAH and BTX) pollutants from flue gas emitted from burning of HFO. This technology is a multipollutant emission control technology that can be applied for treatment of flue gas emitted from coal-, lignite-, and HFO-fired boilers. Other thermal processes such as metallurgy and municipal waste incinerators are potential candidates for this technology application.

  19. An Ideal Molecular Sieve for Acetylene Removal from Ethylene with Record Selectivity and Productivity.

    PubMed

    Li, Bin; Cui, Xili; O'Nolan, Daniel; Wen, Hui-Min; Jiang, Mengdie; Krishna, Rajamani; Wu, Hui; Lin, Rui-Biao; Chen, Yu-Sheng; Yuan, Daqiang; Xing, Huabin; Zhou, Wei; Ren, Qilong; Qian, Guodong; Zaworotko, Michael J; Chen, Banglin

    2017-12-01

    Realization of ideal molecular sieves, in which the larger gas molecules are completely blocked without sacrificing high adsorption capacities of the preferred smaller gas molecules, can significantly reduce energy costs for gas separation and purification and thus facilitate a possible technological transformation from the traditional energy-intensive cryogenic distillation to the energy-efficient, adsorbent-based separation and purification in the future. Although extensive research endeavors are pursued to target ideal molecular sieves among diverse porous materials, over the past several decades, ideal molecular sieves for the separation and purification of light hydrocarbons are rarely realized. Herein, an ideal porous material, SIFSIX-14-Cu-i (also termed as UTSA-200), is reported with ultrafine tuning of pore size (3.4 Å) to effectively block ethylene (C 2 H 4 ) molecules but to take up a record-high amount of acetylene (C 2 H 2 , 58 cm 3 cm -3 under 0.01 bar and 298 K). The material therefore sets up new benchmarks for both the adsorption capacity and selectivity, and thus provides a record purification capacity for the removal of trace C 2 H 2 from C 2 H 4 with 1.18 mmol g -1 C 2 H 2 uptake capacity from a 1/99 C 2 H 2 /C 2 H 4 mixture to produce 99.9999% pure C 2 H 4 (much higher than the acceptable purity of 99.996% for polymer-grade C 2 H 4 ), as demonstrated by experimental breakthrough curves. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Synthesis of Surface-Modified Iron Oxides for the Solvent-Free Recovery of Bacterial Bioactive Compound Prodigiosin and Its Algicidal Activity.

    PubMed

    Arivizhivendhan, K V; Mahesh, M; Boopathy, R; Patchaimurugan, K; Maharaja, P; Swarnalatha, S; Regina Mary, R; Sekaran, G

    2016-09-15

    Prodigiosin (PG) is a bioactive compound produced by several bacterial species. Currently, many technologies are being developed for the production of PG by fermentation processes. However, new challenges are being faced with regard to the production of PG in terms of the recovery and purification steps, owing to the labile nature of PG molecules and the cost of the purification steps. Conventional methods have limitations due to high cost, low reusability, and health hazards. Hence, the present investigation was focused on the development of surface-functionalized magnetic iron oxide ([Fe3O4]F) for solvent-free extraction of bioactive PG from the bacterial fermented medium. Fe3O4 was functionalized with diethanolamine and characterized by FT-IR, diffuse reflectance spectroscopy, thermogravimetric analysis, scanning electron microscopy, and confocal microscopy. The various process parameters, such as contact time, temperature, pH, and mass of Fe3O4, were optimized for the extraction of PG using functionalized Fe3O4. Instrumental analyses confirmed that the PG molecules were cross-linked with functional groups on [Fe3O4]F through van der Waals forces of attraction. PG extracted through Fe3O4 or [Fe3O4]F was separated from the fermentation medium by applying an external electromagnetic field and regenerated for successive reuse cycles. The purity of the extracted PG was characterized by high-performance liquid chromatography, FT-IR, and UV-visible spectroscopy. The iron oxide-diethanolamine-PG cross-linked ([Fe3O4]F-PG) composite matrix effectively deactivates harmful fouling by cyanobacterial growth in water-treatment plants. The present investigation provides the possibility of solvent-free extraction of bacterial bioactive PG from a fermented medium using functionalized magnetic iron oxide.

  1. Development of novel magnetic nanoparticles for hyperthermia cancer therapy

    NASA Astrophysics Data System (ADS)

    Cassim, Shiraz M.; Giustini, Andrew J.; Baker, Ian; Hoopes, P. Jack

    2011-03-01

    Advances in magnetic nanoparticle hyperthermia are opening new doors in cancer therapy. As a standalone or adjuvant therapy this new modality has the opportunity significantly advance thermal medicine. Major advantages of using magnetic magnetite (Fe3O4) nanoparticles are their highly localized power deposition and the fact that the alternating magnetic fields (AMF) used to excite them can penetrate deeply into the body without harmful effect. One limitation, however, which hinders the technology, is the problem of inductive heating of normal tissue by the AMF if the frequency and fields strength are not appropriately matched to the tissue. Restricting AMF amplitude and frequency limits the heat dose which can be selectively applied to cancerous tissue via the magnetic nanoparticle, thus lowering therapeutic effect. In an effort to address this problem, particles with optimized magnetic properties must be developed. Using particles with higher saturation magnetizations and coercivity will enhance hysteresis heating increasing particle power density at milder AMF strengths and frequencies. In this study we used oil in water microemulsions to develop nanoparticles with zero-valent Fe cores and magnetite shells. The superior magnetic properties of zero-valent Fe give these particles the potential for improved SAR over pure magnetite particles. Silane and subsequently dextran have been attached to the particle surface in order to provide a biocompatible surfactant coating. The heating capability of the particles was tested in-vivo using a mouse tumor model. Although we determined that the final stage of synthesis, purification of the dextran coated particles, permits significant corrosion/oxidation of the iron core to hematite, the particles can effectively heat tumor tissue. Improving the purification procedure will allow the generation Fe/Fe3O4 with superior SAR values.

  2. Partitioning in aqueous two-phase systems: Analysis of strengths, weaknesses, opportunities and threats.

    PubMed

    Soares, Ruben R G; Azevedo, Ana M; Van Alstine, James M; Aires-Barros, M Raquel

    2015-08-01

    For half a century aqueous two-phase systems (ATPSs) have been applied for the extraction and purification of biomolecules. In spite of their simplicity, selectivity, and relatively low cost they have not been significantly employed for industrial scale bioprocessing. Recently their ability to be readily scaled and interface easily in single-use, flexible biomanufacturing has led to industrial re-evaluation of ATPSs. The purpose of this review is to perform a SWOT analysis that includes a discussion of: (i) strengths of ATPS partitioning as an effective and simple platform for biomolecule purification; (ii) weaknesses of ATPS partitioning in regard to intrinsic problems and possible solutions; (iii) opportunities related to biotechnological challenges that ATPS partitioning may solve; and (iv) threats related to alternative techniques that may compete with ATPS in performance, economic benefits, scale up and reliability. This approach provides insight into the current status of ATPS as a bioprocessing technique and it can be concluded that most of the perceived weakness towards industrial implementation have now been largely overcome, thus paving the way for opportunities in fermentation feed clarification, integration in multi-stage operations and in single-step purification processes. Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Microfluidic cartridges for DNA purification and genotyping processed in standard laboratory instruments

    NASA Astrophysics Data System (ADS)

    Focke, Maximilian; Mark, Daniel; Stumpf, Fabian; Müller, Martina; Roth, Günter; Zengerle, Roland; von Stetten, Felix

    2011-06-01

    Two microfluidic cartridges intended for upgrading standard laboratory instruments with automated liquid handling capability by use of centrifugal forces are presented. The first microfluidic cartridge enables purification of DNA from human whole blood and is operated in a standard laboratory centrifuge. The second microfluidic catridge enables genotyping of pathogens by geometrically multiplexed real-time PCR. It is operated in a slightly modified off-the-shelf thermal cycler. Both solutions aim at smart and cost-efficient ways to automate work flows in laboratories. The DNA purification cartridge automates all liquid handling steps starting from a lysed blood sample to PCR ready DNA. The cartridge contains two manually crushable glass ampoules with liquid reagents. The DNA yield extracted from a 32 μl blood sample is 192 +/- 30 ng which corresponds to 53 +/- 8% of a reference extraction. The genotyping cartridge is applied to analyse isolates of the multi-resistant Staphyloccus aureus (MRSA) by real-time PCR. The wells contain pre-stored dry reagents such as primers and probes. Evaluation of the system with 44 genotyping assays showed a 100% specificity and agreement with the reference assays in standard tubes. The lower limit of detection was well below 10 copies of DNA per reaction.

  4. Stabilization and immobilization of aquaporin reconstituted lipid vesicles for water purification.

    PubMed

    Sun, Guofei; Chung, Tai-Shung; Jeyaseelan, Kandiah; Armugam, Arunmozhiarasi

    2013-02-01

    Aquaporins are water channel proteins in biological membranes that have extraordinary water permeability and selectivity. In this work, we have demonstrated that one of their family members, AquaporinZ (AqpZ), can be possibly applied in a pressure-driven water purification process. A nanofiltration membrane was designed and fabricated by immobilization of AqpZ-reconstituted liposomes on a polydopamine (PDA) coated microporous membrane. Amine-functionalized proteoliposomes were first deposited via gentle vacuum suction and subsequently conjugated on the PDA layer via an amine-catechol adduct formation. Due to the existence of a polymer network within the lipid bilayers, the membrane could sustain hydraulic pressure of 5 bar as well as the strong surface agitation in nanofiltration tests, indicating a relatively stable membrane structure. In comparison with membrane without AqpZ incorporation, the membrane with AqpZ-to-lipid weight ratio of 1:100 increased the water flux by 65% with enhanced NaCl and MgCl(2) rejections of 66.2% and 88.1%, respectively. With AqpZ incorporation, the vesicle immobilized membrane exhibits a promising strategy for high productivity water purification. Copyright © 2012 Elsevier B.V. All rights reserved.

  5. Identifying Protein-protein Interaction in Drosophila Adult Heads by Tandem Affinity Purification (TAP)

    PubMed Central

    Tian, Xiaolin; Zhu, Mingwei; Li, Long; Wu, Chunlai

    2013-01-01

    Genetic screens conducted using Drosophila melanogaster (fruit fly) have made numerous milestone discoveries in the advance of biological sciences. However, the use of biochemical screens aimed at extending the knowledge gained from genetic analysis was explored only recently. Here we describe a method to purify the protein complex that associates with any protein of interest from adult fly heads. This method takes advantage of the Drosophila GAL4/UAS system to express a bait protein fused with a Tandem Affinity Purification (TAP) tag in fly neurons in vivo, and then implements two rounds of purification using a TAP procedure similar to the one originally established in yeast1 to purify the interacting protein complex. At the end of this procedure, a mixture of multiple protein complexes is obtained whose molecular identities can be determined by mass spectrometry. Validation of the candidate proteins will benefit from the resource and ease of performing loss-of-function studies in flies. Similar approaches can be applied to other fly tissues. We believe that the combination of genetic manipulations and this proteomic approach in the fly model system holds tremendous potential for tackling fundamental problems in the field of neurobiology and beyond. PMID:24335807

  6. Production, Purification, and Identification of Cholest-4-en-3-one Produced by Cholesterol Oxidase from Rhodococcus sp. in Aqueous/Organic Biphasic System.

    PubMed

    Wu, Ke; Li, Wei; Song, Jianrui; Li, Tao

    2015-01-01

    Cholest-4-en-3-one has positive uses against obesity, liver disease, and keratinization. It can be applied in the synthesis of steroid drugs as well. Most related studies are focused on preparation of cholest-4-en-3-one by using whole cells as catalysts, but production of high-quality cholest-4-en-3-one directly from cholesterol oxidase (COD) using an aqueous/organic two-phase system has been rarely explored. This study set up an enzymatic reaction system to produce cholest-4-en-3-one. We developed and optimized the enzymatic reaction system using COD from COX5-6 (a strain of Rhodococcus) instead of whole-cell biocatalyst. This not only simplifies and accelerates the production but also benefits the subsequent separation and purification process. Through extraction, washing, evaporation, column chromatography, and recrystallization, we got cholest-4-en-3-one with purity of 99.78%, which was identified by nuclear magnetic resonance, mass spectroscopy, and infrared spectroscopy. In addition, this optimized process of cholest-4-en-3-one production and purification can be easily scaled up for industrial production, which can largely decrease the cost and guarantee the purity of the product.

  7. Low cost thermoformed solar still water purifier for D&E countries

    NASA Astrophysics Data System (ADS)

    Flendrig, L. M.; Shah, B.; Subrahmaniam, N.; Ramakrishnan, V.

    IntroductionSolar distillation mimics nature’s hydrologic water cycle by purifying water through evaporation (using solar energy) and condensation (rain). It is one of the most basic purification systems available today to obtain high quality drinking water and can remove non-volatile contamination from almost any water source. This low-tech technology should therefore be ideally suited for developing and emerging countries where sun shines in abundance. In the past century numerous designs have been realised with footprints ranging from 0.5 m 2 to thousands of square meters. Despite all efforts, this intriguing technology has not been applied widely yet. Among the challenges that remain are: (1) its low yield, (2) obtaining local commitment to operate/maintain large scale systems properly, and (3) relatively high initial investment costs. The objective of this study has been to address challenges 1 and 3 by using standard plastic thermoforming technology to realize a small scale single slope solar still for personal use (2-4 l per day) with adequate efficiency and at low production costs. Materials and methodsThe solar still consists of two parts: a basin that holds the dirty water and a transparent tilted cover onto which the clean water vapour can condense. The basin has a footprint of 1.34 m 2 and is made of a 3 mm thick sheet of black high-density polyethylene (HDPE) which is thermoformed using standard equipment for making fish-ponds. This allows for the incorporation of detailed features, like reinforcements and a clean-water collection gutter, at no extra cost. The transparent cover is made of UV stabilised low-density PE-foil which is under a slope of 10° to transport condensed water droplets to the lower located collection gutter. Throughput and purification performance were evaluated in duplicate at our Bangalore R&D facilities in India, over a short term (5 day) period. Solar radiation was measured using a Pyranometer. The system was loaded with 40 l of laundry rinse water. ResultsAt an average solar radiation of 12.95 MJ/day/m 2 the average yield of purified water was 3 l/day. This resulted in a calculated overall system efficiency of 39%. Purification performance (contaminated versus purified water) of the solar still loaded with the most contaminated water source was: Total dissolved solids (TDS) from 2925 ppm to 40 ppm, pH from 9.6 to 5.5, conductivity from 6130 mS/cm to 26 mS/cm, turbidity from 394 NTU to 0.4 NTU, total viable count (TVC) from 314 million cfu/ml to <10 cfu/ml. ConclusionThermoforming allowed for the realisation of a single slope solar still that can sustainably produce high quality drinking water at point of use from waste water with an above average efficiency and at a manufacturing price (in The Netherlands) of below €25-per system. Next step should focus on a long term evaluation (months, instead of days) to access the full potential of the solar still to produce safe drinking water at point of use in an economical and reliable way.

  8. Cephalopod-Inspired Reflectin-Based Photonic Devices

    NASA Astrophysics Data System (ADS)

    Phan, Long

    Cephalopods are known as the chameleons of the sea due to their remarkable camouflage abilities. They can rapidly and accurately tune their skin's coloration, pattern, and texture to blend into the surrounding environment. This dynamic camouflage capability stems from their transparent dermis/epidermis and the optically-active, protein-based nanostructures found in embedded skin cells known as leucophores, chromatophores, and iridophores. Respectively, these cells provide a high contrast reflective white background, mechanically actuated pigmented pixels, and chemically actuated Bragg reflectors that function in concert to modulate incident visible light. Considerable effort has been devoted to understanding and emulating cephalopod camouflage abilities in the visible region of the electromagnetic spectrum, but few studies have attempted to translate these principles to the infrared region for nighttime stealth applications. Thus, the fabrication of bio-inspired infrared-reflective devices for infrared camouflage remains an unexplored area of research. To address this challenge, we have developed a high-throughput strategy for the gram-scale production, purification, and self-assembly of a unique cephalopod structural protein, reflectin. We eliminate time-consuming and costly steps commonly used in protein expression and purification and instead replace them with rapid, sequential filtrations all while retaining high purity (>99%). Using this reflectin protein, we fabricate dynamically tunable biomimetic camouflage coatings with relevance to industrial and military applications. We demonstrate reversible control of reflectin film coloration shifts over a range of 1,200 nm from the visible into the near infrared using an acid vapor stimulus. We then coat reflectin on flexible, transparent substrates that can adhere to arbitrary surfaces, and modulate the film reflectance by mechanical strain or applied heat. Finally, we prove electrical actuation can also induce reversible color change in our films based on the applied bias. Together, our findings represent a key step towards the development of wearable biomimetic color and shapeshifting technologies that utilize diverse means of actuation. Future biophysical and materials studies lending insight into the tunability of reflectin-based Bragg reflector structures and textured reflectin surfaces could provide additional methods to enhance overall film brightness, angle-dependence, and color modulation for advanced camouflage applications.

  9. Tanks and Tank Troops

    DTIC Science & Technology

    1982-03-01

    dis- tance of up to 1,200 meters in moonlight . Effective range is substantially reduced with weaker starlight. Night vision devices with...by purification treatment and employment of corrosion inhibitors; by applying protective films to the vulnerable surface (chemically passive, oil...elaborated dynamic stereotype , tanker efficiency Improves, as do march performance Indicators. The duration of this period Is approximately 24 hours

  10. Characterization of biobased polyurethane foams employing lignin fractionated from microwave liquefied switchgrass

    Treesearch

    Xingyan Huang; Cornelis F. De Hoop; Jiulong Xie; Chung-Yun Hse; Jinqiu Qi; Tingxing Hu

    2017-01-01

    Lignin samples fractionated from microwave liquefied switchgrass were applied in the preparation of semirigid polyurethane (PU) foams without purification.The objective of this study was to elucidate the influence of lignin in the PU matrix on themorphological, chemical, mechanical, and thermal properties of thePU foams.The scanning electron microscopy (SEM) images...

  11. [Identification of the interacting proteins with S100A8 or S100A9 by affinity purification and mass spectrometry].

    PubMed

    Wang, Jing; Zhang, Xuemei; Li, Zheng; Li, Xiayu; Ma, Jian; Shen, Shourong

    2017-04-28

    To identify the interacting proteins with S100A8 or S100A9 in HEK293 cell line by flag-tag affinity purification and liquid chromatography mass spectrometry/mass spectrometry (LC-MS/MS).
 Methods: The p3×Flag-CMV-S100A8 and p3×Flag-CMV-S100A9 expression vectors were constructed by inserting S100A8 or S100A9 coding sequence. The recombinant plasmids were then transfected into HEK293 cells. Affinity purification and LC-MS/MS were applied to identify the proteins interacting with S100A8 or S100A9. Bioinformatics analysis was used to seek the gene ontology of the interacting proteins. Co-immunoprecipitation (Co-IP) was applied to confirm the proteins interacted with S100A8 or S100A9.
 Results: Fourteen proteins including pyruvate kinase, muscle (PKM), nucleophosmin (NPM1) and eukaryotic translation initiation factor 5A (EIF5A), which potentially interacted with S100A8, were successfully identified by Flag-tag affinity purification followed by LC-MS/MS analysis. Six proteins, such as tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein epsilon (14-3-3ε) and PKM, which potentially interacted with S100A9, were successfully identified. Gene ontology analysis of the identified proteins suggested that proteins interacted with S100A8 or S100A9 were involved in several biological pathways, including canonical glycolysis, positive regulation of NF-κB transcription factor activity, negative regulation of apoptotic process, cell-cell adhesion, etc. Co-IP experiment confirmed that PKM2 can interact with both S100A8 and S100A9, and 14-3-3ε can interact with S100A8.
 Conclusion: PKM2 is identified to interact with both S100A8 and S100A9, while 14-3-3ε can interact with S100A9. These results may provide a new clue for the role of S100A8 or S100A9 in the progression of colitis-associated colorectal cancer.

  12. Nuclear Weapons. National Nuclear Security Administration’s Plans for Its Uranium Processing Facility Should Better Reflect Funding Estimates and Technology Readiness

    DTIC Science & Technology

    2010-11-01

    metal. Recovery extraction centrifugal contactors A process that uses solvent to extract uranium for purposes of purification. Agile machining A...extraction centrifugal contactors 5 6 Yes 6 No Agile machining 5 5 No 6 No Chip management 5 6 Yes 6 No Special casting 3 6 Yes 6 No Source: GAO

  13. PEL Staff Together for the First Time | Poster

    Cancer.gov

    By Ashley DeVine, Staff Writer John-Paul Denson and Troy Taylor of the Protein Expression Laboratory (PEL) used to pack liters of Escherichia coli lysates on ice, put them in the back of a microvan, and drive across campus to deliver the samples for protein purification. Now that all PEL staff members are working under the same roof at the Advanced Technology Research Facility

  14. On the Recent Use of Membrane Technology for Olive Mill Wastewater Purification

    PubMed Central

    Ochando-Pulido, Javier Miguel; Martinez-Ferez, Antonio

    2015-01-01

    Many reclamation treatments as well as integrated processes for the purification of olive mill wastewaters (OMW) have already been proposed and developed but not led to completely satisfactory results, principally due to complexity or cost-ineffectiveness. The olive oil industry in its current status, composed of little and dispersed factories, cannot stand such high costs. Moreover, these treatments are not able to abate the high concentration of dissolved inorganic matter present in these highly polluted effluents. In the present work, a review on the actual state of the art concerning the treatment and disposal of OMW by membranes is addressed, comprising microfiltration (MF), ultrafiltration (UF), nanofiltration (NF), and reverse osmosis (RO), as well as membrane bioreactors (MBR) and non-conventional membrane processes such as vacuum distillation (VD), osmotic distillation (OD) and forward osmosis (FO). Membrane processes are becoming extensively used to replace many conventional processes in the purification of water and groundwater as well as in the reclamation of wastewater streams of very diverse sources, such as those generated by agro-industrial activities. Moreover, a brief insight into inhibition and control of fouling by properly-tailored pretreatment processes upstream the membrane operation and the use of the critical and threshold flux theories is provided. PMID:26426062

  15. A Practical Cryogen-Free CO2 Purification and Freezing Technique for Stable Isotope Analysis.

    PubMed

    Sakai, Saburo; Matsuda, Shinichi

    2017-04-18

    Since isotopic analysis by mass spectrometry began in the early 1900s, sample gas for light-element isotopic measurements has been purified by the use of cryogens and vacuum-line systems. However, this conventional purification technique can achieve only certain temperatures that depend on the cryogens and can be sustained only as long as there is a continuous cryogen supply. Here, we demonstrate a practical cryogen-free CO 2 purification technique using an electrical operated cryocooler for stable isotope analysis. This approach is based on portable free-piston Stirling cooling technology and controls the temperature to an accuracy of 0.1 °C in a range from room temperature to -196 °C (liquid-nitrogen temperature). The lowest temperature can be achieved in as little as 10 min. We successfully purified CO 2 gas generated by carbonates and phosphoric acid reaction and found its sublimation point to be -155.6 °C at 0.1 Torr in the vacuum line. This means that the temperature required for CO 2 trapping is much higher than the liquid-nitrogen temperature. Our portable cooling system offers the ability to be free from the inconvenience of cryogen use for stable isotope analysis. It also offers a new cooling method applicable to a number of fields that use gas measurements.

  16. Graphene-Based Standalone Solar Energy Converter for Water Desalination and Purification.

    PubMed

    Yang, Yang; Zhao, Ruiqi; Zhang, Tengfei; Zhao, Kai; Xiao, Peishuang; Ma, Yanfeng; Ajayan, Pulickel M; Shi, Gaoquan; Chen, Yongsheng

    2018-01-23

    Harvesting solar energy for desalination and sewage treatment has been considered as a promising solution to produce clean water. However, state-of-the-art technologies often require optical concentrators and complicated systems with multiple components, leading to poor efficiency and high cost. Here, we demonstrate an extremely simple and standalone solar energy converter consisting of only an as-prepared 3D cross-linked honeycomb graphene foam material without any other supporting components. This simple all-in-one material can act as an ideal solar thermal converter capable of capturing and converting sunlight into heat, which in turn can distill water from various water sources into steam and produce purified water under ambient conditions and low solar flux with very high efficiency. High specific water production rate of 2.6 kg h -1 m -2 g -1 was achieved with near ∼87% under 1 sun intensity and >80% efficiency even under ambient sunlight (<1 sun). This scalable sheet-like material was used to obtain pure drinkable water from both seawater and sewage water under ambient conditions. Our results demonstrate a competent monolithic material platform providing a paradigm change in water purification by using a simple, point of use, reusable, and low-cost solar thermal water purification system for a variety of environmental conditions.

  17. The Electrolytic Effect on the Catalytic Degradation of Dye and Nitrate Ion by New Ceramic Beads of Natural Minerals and TiO2

    NASA Astrophysics Data System (ADS)

    Sata, Akiyoshi; Sakai, Takako; Goto, Yusuke; Ohta, Toshiyuki; Hayakawa, Katumitu

    2007-05-01

    We have developed a new hybrid ceramic material "Taiyo" as a water processing catalyst. The porous ceramic has a core-shell structure. It decolorized completely the dye solutions as well as the wastewater output after primary water processing by microorganism in a pig farm. This new material showed the acceleration of water purification by applying electric voltage. The degradation of dyes and pig urine output from the primary treatments was accelerated by applying voltage. Nitrate in underground water was also decomposed only by applying voltage, while it was not decomposed without voltage.

  18. Nucleic acid programmable protein array a just-in-time multiplexed protein expression and purification platform.

    PubMed

    Qiu, Ji; LaBaer, Joshua

    2011-01-01

    Systematic study of proteins requires the availability of thousands of proteins in functional format. However, traditional recombinant protein expression and purification methods have many drawbacks for such study at the proteome level. We have developed an innovative in situ protein expression and capture system, namely NAPPA (nucleic acid programmable protein array), where C-terminal tagged proteins are expressed using an in vitro expression system and efficiently captured/purified by antitag antibodies coprinted at each spot. The NAPPA technology presented in this chapter enable researchers to produce and display fresh proteins just in time in a multiplexed high-throughput fashion and utilize them for various downstream biochemical researches of interest. This platform could revolutionize the field of functional proteomics with it ability to produce thousands of spatially separated proteins in high density with narrow dynamic rand of protein concentrations, reproducibly and functionally. Copyright © 2011 Elsevier Inc. All rights reserved.

  19. RNA Crystallization

    NASA Technical Reports Server (NTRS)

    Golden, Barbara L.; Kundrot, Craig E.

    2003-01-01

    RNA molecules may be crystallized using variations of the methods developed for protein crystallography. As the technology has become available to syntheisize and purify RNA molecules in the quantities and with the quality that is required for crystallography, the field of RNA structure has exploded. The first consideration when crystallizing an RNA is the sequence, which may be varied in a rational way to enhance crystallizability or prevent formation of alternate structures. Once a sequence has been designed, the RNA may be synthesized chemically by solid-state synthesis, or it may be produced enzymatically using RNA polymerase and an appropriate DNA template. Purification of milligram quantities of RNA can be accomplished by HPLC or gel electrophoresis. As with proteins, crystallization of RNA is usually accomplished by vapor diffusion techniques. There are several considerations that are either unique to RNA crystallization or more important for RNA crystallization. Techniques for design, synthesis, purification, and crystallization of RNAs will be reviewed here.

  20. A non-chromatographic protein purification strategy using Src 3 homology domains as generalized capture domains.

    PubMed

    Kim, Heejae; Chen, Wilfred

    2016-09-20

    Protein purification using inverse phase transition of elastin-like polypeptide (ELP) domains is a useful alternative to chromatography. Genetic fusions of ELP domains to various proteins have the ability to reversibly transition between soluble monomers and micron-sized aggregates and this has been used to selectively purify many ELP fusions. Affinity domains can enhance this technology by using specific protein binding domains to enable ELP mediated affinity capture (EMAC) of proteins of interest (POI) that have been fused to corresponding affinity ligands. In this paper, we highlight the use of Src homology 3 (SH3) domains and corresponding peptide ligands in EMAC that have differential binding affinities towards SH3 for efficient capture and elution of proteins. Furthermore, differences between capture and elution of a monomeric and a multimeric protein were also studied. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Improvements in purification of silver nanowires by decantation and fabrication of flexible transparent electrodes. Application to capacitive touch sensors.

    PubMed

    Mayousse, Céline; Celle, Caroline; Moreau, Eléonore; Mainguet, Jean-François; Carella, Alexandre; Simonato, Jean-Pierre

    2013-05-31

    Transparent flexible electrodes made of metallic nanowires, and in particular silver nanowires (AgNWs), appear as an extremely promising alternative to transparent conductive oxides for future optoelectronic devices. Though significant progresses have been made the last few years, there is still some room for improvement regarding the synthesis of high quality silver nanowire solutions and fabrication process of high performance electrodes. We show that the commonly used purification process can be greatly simplified through decantation. Using this process it is possible to fabricate flexible electrodes by spray coating with sheet resistance lower than 25 Ω sq⁻¹ at 90% transparency in the visible spectrum. These electrodes were used to fabricate an operative transparent flexible touch screen. To our knowledge this is the first reported AgNW based touch sensor relying on capacitive technology.

  2. Reverse micellar extraction of bromelain from pineapple peel--Effect of surfactant structure.

    PubMed

    Wan, Jing; Guo, Jingjing; Miao, Zhitong; Guo, Xia

    2016-04-15

    Pineapple peel is generally disposed or used as compost. This study was focused on extracting bromelain from pineapple peel by using reverse micelles. It was found that gemini surfactant C12-8-C12·2Br (octamethylene-α,ω-bis(dimethyldodecylammonium bromide)) showed distinctive advantage over its monomeric counterpart DTAB (dodecyl trimethyl ammonium bromide); under optimized condition, the bromelain extracted with C12-8-C12·2Br reverse micelle had an activity recovery of 163% and a purification fold of 3.3, while when using DTAB reverse micelle, the activity recovery was 95% and the purification fold was 1.7. Therefore, the spacer of gemini surfactant should play a positive role in bromelain extraction and may suggest the potential of gemini surfactant in protein separation since it has been so far rarely used in relative experiments or technologies. Copyright © 2015 Elsevier Ltd. All rights reserved.

  3. Analysis of gas membrane ultra-high purification of small quantities of mono-isotopic silane

    DOE PAGES

    de Almeida, Valmor F.; Hart, Kevin J.

    2017-01-03

    A small quantity of high-value, crude, mono-isotopic silane is a prospective gas for a small-scale, high-recovery, ultra-high membrane purification process. This is an unusual application of gas membrane separation for which we provide a comprehensive analysis of a simple purification model. The goal is to develop direct analytic expressions for estimating the feasibility and efficiency of the method, and guide process design; this is only possible for binary mixtures of silane in the dilute limit which is a somewhat realistic case. In addition, analytic solutions are invaluable to verify numerical solutions obtained from computer-aided methods. Hence, in this paper wemore » provide new analytic solutions for the purification loops proposed. Among the common impurities in crude silane, methane poses a special membrane separation challenge since it is chemically similar to silane. Other potential problematic compounds are: ethylene, diborane and ethane (in this order). Nevertheless, we demonstrate, theoretically, that a carefully designed membrane system may be able to purify mono-isotopic, crude silane to electronics-grade level in a reasonable amount of time and expenses. We advocate a combination of membrane materials that preferentially reject heavy impurities based on mobility selectivity, and light impurities based on solubility selectivity. We provide estimates for the purification of significant contaminants of interest. In this study, we suggest cellulose acetate and polydimethylsiloxane as examples of membrane materials on the basis of limited permeability data found in the open literature. We provide estimates on the membrane area needed and priming volume of the cell enclosure for fabrication purposes when using the suggested membrane materials. These estimates are largely theoretical in view of the absence of reliable experimental data for the permeability of silane. And finally, future extension of this work to the non-dilute limit may apply to the recovery of silane from rejected streams of natural silicon semi-conductor processes.« less

  4. Rapid One-step Enzymatic Synthesis and All-aqueous Purification of Trehalose Analogues.

    PubMed

    Meints, Lisa M; Poston, Anne W; Piligian, Brent F; Olson, Claire D; Badger, Katherine S; Woodruff, Peter J; Swarts, Benjamin M

    2017-02-17

    Chemically modified versions of trehalose, or trehalose analogues, have applications in biology, biotechnology, and pharmaceutical science, among other fields. For instance, trehalose analogues bearing detectable tags have been used to detect Mycobacterium tuberculosis and may have applications as tuberculosis diagnostic imaging agents. Hydrolytically stable versions of trehalose are also being pursued due to their potential for use as non-caloric sweeteners and bioprotective agents. Despite the appeal of this class of compounds for various applications, their potential remains unfulfilled due to the lack of a robust route for their production. Here, we report a detailed protocol for the rapid and efficient one-step biocatalytic synthesis of trehalose analogues that bypasses the problems associated with chemical synthesis. By utilizing the thermostable trehalose synthase (TreT) enzyme from Thermoproteus tenax, trehalose analogues can be generated in a single step from glucose analogues and uridine diphosphate glucose in high yield (up to quantitative conversion) in 15-60 min. A simple and rapid non-chromatographic purification protocol, which consists of spin dialysis and ion exchange, can deliver many trehalose analogues of known concentration in aqueous solution in as little as 45 min. In cases where unreacted glucose analogue still remains, chromatographic purification of the trehalose analogue product can be performed. Overall, this method provides a "green" biocatalytic platform for the expedited synthesis and purification of trehalose analogues that is efficient and accessible to non-chemists. To exemplify the applicability of this method, we describe a protocol for the synthesis, all-aqueous purification, and administration of a trehalose-based click chemistry probe to mycobacteria, all of which took less than 1 hour and enabled fluorescence detection of mycobacteria. In the future, we envision that, among other applications, this protocol may be applied to the rapid synthesis of trehalose-based probes for tuberculosis diagnostics. For instance, short-lived radionuclide-modified trehalose analogues (e.g., 18 F-modified trehalose) could be used for advanced clinical imaging modalities such as positron emission tomography-computed tomography (PET-CT).

  5. Analysis of gas membrane ultra-high purification of small quantities of mono-isotopic silane

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    de Almeida, Valmor F.; Hart, Kevin J.

    A small quantity of high-value, crude, mono-isotopic silane is a prospective gas for a small-scale, high-recovery, ultra-high membrane purification process. This is an unusual application of gas membrane separation for which we provide a comprehensive analysis of a simple purification model. The goal is to develop direct analytic expressions for estimating the feasibility and efficiency of the method, and guide process design; this is only possible for binary mixtures of silane in the dilute limit which is a somewhat realistic case. In addition, analytic solutions are invaluable to verify numerical solutions obtained from computer-aided methods. Hence, in this paper wemore » provide new analytic solutions for the purification loops proposed. Among the common impurities in crude silane, methane poses a special membrane separation challenge since it is chemically similar to silane. Other potential problematic compounds are: ethylene, diborane and ethane (in this order). Nevertheless, we demonstrate, theoretically, that a carefully designed membrane system may be able to purify mono-isotopic, crude silane to electronics-grade level in a reasonable amount of time and expenses. We advocate a combination of membrane materials that preferentially reject heavy impurities based on mobility selectivity, and light impurities based on solubility selectivity. We provide estimates for the purification of significant contaminants of interest. In this study, we suggest cellulose acetate and polydimethylsiloxane as examples of membrane materials on the basis of limited permeability data found in the open literature. We provide estimates on the membrane area needed and priming volume of the cell enclosure for fabrication purposes when using the suggested membrane materials. These estimates are largely theoretical in view of the absence of reliable experimental data for the permeability of silane. And finally, future extension of this work to the non-dilute limit may apply to the recovery of silane from rejected streams of natural silicon semi-conductor processes.« less

  6. Simple and rapid purification of pediocin PA-1 from Pediococcus pentosaceous NCDC 273 suitable for industrial application.

    PubMed

    Vijay Simha, B; Sood, S K; Kumariya, Rashmi; Garsa, Anita Kumari

    2012-10-12

    The use of pediocins as food additives or drugs requires a simple and rapid method by which large quantities of homogeneous pediocin are produced at industrial level. Two centrifugation steps required during initial stages of purification i.e. separation of cells from fermentation broth and collection of precipitates after ammonium sulphate precipitation are the major bottlenecks for their large scale purification. In the present work, pediocin production by a new a dairy strain, Pediococcus pentosaceous NCDC 273 (identical to pediocin PA-1 at nucleotide sequence level), was found to be optimum at initial pH of 6.0 and 7.0 of basal MRS supplemented with 20 g/l of glucose or lactose at 20 and 24 h, respectively. Immobilization of cells through entrapment in alginate-xanthan gum gel beads with chitosan coating resulted in negligible cell release during fermentation. Thus, the cell free extract was directly collected through decantation, avoiding the need of centrifugation step at this stage. Subsequent ammonium sulphate precipitation at isoelectric point of pediocin PA-1 (8.85), using magnetic stirrer at high speed (approx. 1200 rpm), resulted in forceful deposition of precipitates on the wall of precipitation beaker allowing their collection using a spatula, avoiding centrifugation step at this stage also. Further purification using cation-exchange chromatography resulted in yield of 134.4% with more than 320 fold purification with the specific activity of 19×10⁵ AU/mg. The collection of single peak of pediocin at 41.9min in RP-HPLC, overlapping with standard pediocin PA-1, resulted in yield of 1.15 μg from 20 μl of sample applied. The overlapping of RP-HPLC peak and SDS-PAGE band corresponding to 4.6 kDa, confirmed the purity and identity of pediocin 273 as pediocin PA-1. Copyright © 2012 Elsevier GmbH. All rights reserved.

  7. Simultaneous micronization and purification of bioactive fraction by supercritical antisolvent technology

    PubMed Central

    Hiendrawan, Stevanus; Veriansyah, Bambang; Widjojokusumo, Edward; Tjandrawinata, Raymond R.

    2017-01-01

    Simultaneous micronization and purification of DLBS3233 bioactive fraction, a combination of two Indonesian herbals Lagerstroemia speciosa and Cinnamomum burmannii has been successfully performed via supercritical anti-solvent (SAS) technology. The objective of the present study was to investigate the effectiveness of SAS technology to micronize and reduce coumarin content of DLBS3233. The effects of four SAS process parameters, i.e. pressure, temperature, concentration and solution flow rate on particle formation were investigated. In SAS process, DLBS3233 was dissolved in dimethylformamide (DMF) as the liquid solvent. The solution was then pumped through a nozzle into a chamber simultaneously with supercritical carbon dioxide (SC-CO2) which acts as the anti-solvent, resulting in DLBS3233 precipitation. Physicochemical properties of unprocessed DLBS3233 and SAS-processed DLBS3233 particles were analyzed using scanning electron microscopy (SEM) and high pressure liquid chromatography (HPLC). Total polyphenol content (TPC) was also analyzed. Particles with mean particle size ranging from 0.107±0.028 μm to 0.298±0.138 μm were obtained by varying the process parameters. SAS-processed DLBS3233 particles showed no coumarin content in all experiments studied in this work. Results of TPC analysis revealed no significant change in SAS-processed DLBS3233 particles compared to unprocessed DLBS3233. Nano-sized DLBS3233 particles with no coumarin content have been successfully produced using SAS process. This study demonstrates the ability of SAS for processing herbal medicine in single step process. PMID:28516056

  8. New Approaches to Boar Semen Evaluation, Processing and Improvement.

    PubMed

    Sutovsky, P

    2015-07-01

    The improvement of boar reproductive performance may be the next frontier in reproductive management of swine herd in Unites States, facilitated by better understanding of boar sperm function and by the introduction of new advanced instrumentation in the andrology field. Objective single ejaculate evaluation and individual boar fertility prediction may be possible by introducing automated flow cytometric semen analysis with vital stains (e.g. acrosomal integrity and mito-potential), DNA fragmentation analysis and biomarkers (ubiquitin, PAWP, ALOX15, aggresome) associated with normal or defective sperm phenotypes. Measurement of sperm-produced reactive oxygen species (ROS) is a helpful indicator of normal semen sample. Semen ROS levels could be managed by the addition of ROS-scavenging antioxidants. Alternative energy regeneration substrates and sperm stimulants such as inorganic pyrophosphate and caffeine could increase sperm lifespan in extended semen and within the female reproductive system. Such technology could be combined with timed sperm release in the female reproductive system after artificial insemination. Sperm phenotype analysis by the image-based flow cytometry will go hand in hand with the advancement of swine genomics, linking aberrant sperm phenotype to the fertility influencing gene polymorphisms. Finally, poor-quality ejaculates could be rescued and acceptable ejaculates improved by semen purification methods such as the nanoparticle-based semen purification and magnetic-activated sperm sorting. Altogether, these scientific and technological advances could benefit swine industry, provided that the challenges of new technology adoption, dissemination and cost reduction are met. © 2015 Blackwell Verlag GmbH.

  9. Simultaneous micronization and purification of bioactive fraction by supercritical antisolvent technology.

    PubMed

    Hiendrawan, Stevanus; Veriansyah, Bambang; Widjojokusumo, Edward; Tjandrawinata, Raymond R

    2017-01-01

    Simultaneous micronization and purification of DLBS3233 bioactive fraction, a combination of two Indonesian herbals Lagerstroemia speciosa and Cinnamomum burmannii has been successfully performed via supercritical anti-solvent (SAS) technology. The objective of the present study was to investigate the effectiveness of SAS technology to micronize and reduce coumarin content of DLBS3233. The effects of four SAS process parameters, i.e. pressure, temperature, concentration and solution flow rate on particle formation were investigated. In SAS process, DLBS3233 was dissolved in dimethylformamide (DMF) as the liquid solvent. The solution was then pumped through a nozzle into a chamber simultaneously with supercritical carbon dioxide (SC-CO2) which acts as the anti-solvent, resulting in DLBS3233 precipitation. Physicochemical properties of unprocessed DLBS3233 and SAS-processed DLBS3233 particles were analyzed using scanning electron microscopy (SEM) and high pressure liquid chromatography (HPLC). Total polyphenol content (TPC) was also analyzed. Particles with mean particle size ranging from 0.107±0.028 μ m to 0.298±0.138 μ m were obtained by varying the process parameters. SAS-processed DLBS3233 particles showed no coumarin content in all experiments studied in this work. Results of TPC analysis revealed no significant change in SAS-processed DLBS3233 particles compared to unprocessed DLBS3233. Nano-sized DLBS3233 particles with no coumarin content have been successfully produced using SAS process. This study demonstrates the ability of SAS for processing herbal medicine in single step process.

  10. Solid-phase based on-chip DNA purification through a valve-free stepwise injection of multiple reagents employing centrifugal force combined with a hydrophobic capillary barrier pressure.

    PubMed

    Zhang, Hainan; Tran, Hong Hanh; Chung, Bong Hyun; Lee, Nae Yoon

    2013-03-21

    In this paper, we demonstrate a simple technique for sequentially introducing multiple sample liquids into microchannels driven by centrifugal force combined with a hydrophobic barrier pressure and apply the technique for performing solid-phase based on-chip DNA purification. Three microchannels with varying widths, all equipped with independent sample reservoirs at the inlets, were fabricated on a hydrophobic elastomer, poly(dimethylsiloxane) (PDMS). First, glass beads were packed inside the reaction chamber, and a whole cell containing the DNA extract was introduced into the widest channel by applying centrifugal force for physical adsorption of the DNA onto the glass beads. Next, washing and elution solutions were sequentially introduced into the intermediate and narrowest microchannels, respectively, by gradually increasing the amount of centrifugal force. Through a precise manipulation of the centrifugal force, the DNA adsorbed onto the glass beads was successfully washed and eluted in a continuous manner without the need to introduce each solution manually. A stepwise injection of liquids was successfully demonstrated using multiple ink solutions, the results of which corresponded well with the theoretical analyses. As a practical application, the D1S80 locus of human genomic DNA, which is widely used for forensic purposes, was successfully purified using the microdevice introduced in this study, as demonstrated through successful target amplification. This will pave the way for the construction of a control-free valve system for realizing on-chip DNA purification, which is one of the most labor-intensive and hard-to-miniaturize components, on a greatly simplified and miniaturized platform employing hydrophobic PDMS.

  11. Exploitation of FTA cartridges for the sampling, long-term storage, and DNA-based analyses of plant-parasitic nematodes.

    PubMed

    Marek, Martin; Zouhar, Miloslav; Douda, Ondřej; Maňasová, Marie; Ryšánek, Pavel

    2014-03-01

    The use of DNA-based analyses in molecular plant nematology research has dramatically increased over recent decades. Therefore, the development and adaptation of simple, robust, and cost-effective DNA purification procedures are required to address these contemporary challenges. The solid-phase-based approach developed by Flinders Technology Associates (FTA) has been shown to be a powerful technology for the preparation of DNA from different biological materials, including blood, saliva, plant tissues, and various human and plant microbial pathogens. In this work, we demonstrate, for the first time, that this FTA-based technology is a valuable, low-cost, and time-saving approach for the sampling, long-term archiving, and molecular analysis of plant-parasitic nematodes. Despite the complex structure and anatomical organization of the multicellular bodies of nematodes, we report the successful and reliable DNA-based analysis of nematode high-copy and low-copy genes using the FTA technology. This was achieved by applying nematodes to the FTA cards either in the form of a suspension of individuals, as intact or pestle-crushed nematodes, or by the direct mechanical printing of nematode-infested plant tissues. We further demonstrate that the FTA method is also suitable for the so-called "one-nematode-assay", in which the target DNA is typically analyzed from a single individual nematode. More surprisingly, a time-course experiment showed that nematode DNA can be detected specifically in the FTA-captured samples many years after initial sampling occurs. Collectively, our data clearly demonstrate the applicability and the robustness of this FTA-based approach for molecular research and diagnostics concerning phytonematodes; this research includes economically important species such as the stem nematode (Ditylenchus dipsaci), the sugar beet nematode (Heterodera schachtii), and the Northern root-knot nematode (Meloidogyne hapla).

  12. Recent advances in nanomaterials for water protection and monitoring.

    PubMed

    Das, Rasel; Vecitis, Chad D; Schulze, Agnes; Cao, Bin; Ismail, Ahmad Fauzi; Lu, Xianbo; Chen, Jiping; Ramakrishna, Seeram

    2017-11-13

    The efficient handling of wastewater pollutants is a must, since they are continuously defiling limited fresh water resources, seriously affecting the terrestrial, aquatic, and aerial flora and fauna. Our vision is to undertake an exhaustive examination of current research trends with a focus on nanomaterials (NMs) to considerably improve the performance of classical wastewater treatment technologies, e.g. adsorption, catalysis, separation, and disinfection. Additionally, NM-based sensor technologies are considered, since they have been significantly used for monitoring water contaminants. We also suggest future directions to inform investigators of potentially disruptive NM technologies that have to be investigated in more detail. The fate and environmental transformations of NMs, which need to be addressed before large-scale implementation of NMs for water purification, are also highlighted.

  13. Aldehyde-functionalized porous nanocellulose for effective removal of heavy metal ions from aqueous solutions

    Treesearch

    C. Yao; F. Wang; Z. Cai; X. Wang

    2016-01-01

    Nanoscale sorption is a promising strategy for catalyst and purification system design. In this paper, cellulose nanofibrils (CNFs) were densely attached with aldehyde functional groups on the surface via a mild periodate oxidation process, and then applied as mesoporous sorbents to remove Cu(II) and Pb(II) from aqueous solutions. In the studied concentration range (0....

  14. Purification and characterization of a collagenase from Penicillium sp. UCP 1286 by polyethylene glycol-phosphate aqueous two-phase system.

    PubMed

    de Albuquerque Wanderley, Maria Carolina; Wanderley Duarte Neto, José Manoel; Campos Albuquerque, Wendell Wagner; de Araújo Viana Marques, Daniela; de Albuquerque Lima, Carolina; da Cruz Silvério, Sara Isabel; de Lima Filho, José Luiz; Couto Teixeira, José António; Porto, Ana Lúcia Figueiredo

    2017-05-01

    Collagenases are proteolytic enzymes capable of degrading both native and denatured collagen, reported to be applied in industrial, medical and biotechnological sectors. Liquid-liquid extraction using aqueous two-phase system (ATPS) is one of the most promising bioseparation techniques, which can substitute difficult solid-liquid separation processes, offering many advantages over conventional methods including low-processing time, low-cost material and low-energy consumption. The collagenase produced by Penicillium sp. UCP 1286 showed a stronger affinity for the bottom salt-rich phase, where the highest levels of collagenolytic activity were observed at the center point runs, using 15.0% (w/w) PEG 3350 g/mol and 12.5% (w/w) phosphate salt at pH 7.0 and concentration. The enzyme was characterized by thermal stability, pH tolerance and effect of inhibitors, showing optimal collagenolytic activity at 37 °C and pH 9.0 and proved to be a serine protease. ATPS showed high efficiency in the collagenase purification, confirmed by a single band in SDS/PAGE, and can in fact be applied as a quick and inexpensive alternative method. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. The experimental study of the effect of microwave on the physical properties of multi-walled carbon nanotubes

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Haque, A.K.M. Mahmudul; Oh, Geum Seok; Kim, Taeoh

    Highlights: • We study the microwave effect on the multi-walled carbon nanotubes (MWCNTs). • We examine the non uniform heating effect on the physical structure of MWCNTs. • We examine the purification of MWCNTs by microwave. • We analyze the thermal characteristics of microwave treated MWCNTs. - Abstract: This paper reports the effect of microwave on the physical properties of multi-walled carbon nanotubes (MWCNTs) where different power levels of microwave were applied on MWCNTs in order to apprehend the effect of microwave on MWCNTs distinctly. A low energy ball milling in aqueous circumstance was also applied on both MWCNTs andmore » microwave treated MWCNTs. Temperature profile, morphological analysis by field emission scanning electron microscopy (FESEM), defect analysis by Raman spectroscopy, thermal conductivity, thermal diffusivity as well as heat transfer coefficient enhancement ratio were studied which expose some strong witnesses of the effect of microwave on the both purification and dispersion properties of MWCNTs in base fluid distilled water. The highest thermal conductivity enhancement (6.06% at 40 °C) of MWCNTs based nanofluid is achieved by five minutes microwave treatment as well as wet grinding at 500 rpm for two hours.« less

  16. Isolation and Purification of Biotechnological Products

    NASA Astrophysics Data System (ADS)

    Hubbuch, Jürgen; Kula, Maria-Regina

    2007-05-01

    The production of modern pharma proteins is one of the most rapid growing fields in biotechnology. The overall development and production is a complex task ranging from strain development and cultivation to the purification and formulation of the drug. Downstream processing, however, still accounts for the major part of production costs. This is mainly due to the high demands on purity and thus safety of the final product and results in processes with a sequence of typically more than 10 unit operations. Consequently, even if each process step would operate at near optimal yield, a very significant amount of product would be lost. The majority of unit operations applied in downstream processing have a long history in the field of chemical and process engineering; nevertheless, mathematical descriptions of the respective processes and the economical large-scale production of modern pharmaceutical products are hampered by the complexity of the biological feedstock, especially the high molecular weight and limited stability of proteins. In order to develop new operational steps as well as a successful overall process, it is thus a necessary prerequisite to develop a deeper understanding of the thermodynamics and physics behind the applied processes as well as the implications for the product.

  17. Organic wastes decomposition technology, perspective for long-term autonomous missions

    NASA Astrophysics Data System (ADS)

    Viacheslav, Ilyin; Korshunov, Denis; Mardanov, Robert; Starkova, Lyubov; Deshevaya, Elena; Smirnov, Igor

    At present time there is no large problem in waste management in ISS space flight conditions, since spacecrafts "Progress" is used for it's removal from orbital station and the wastes burns in dense layers of Earth's atmosphere. However such method does not approach for far inter-planetary flights since interplanetary quarantine desires do not allow to deposit contaminated wastes outside the spacecraft. Essential part of wastes is formed by disposed means of personal hygiene and greenhouse wastes which are not safe from sanitary-epidemiological aspect. Above mentioned materials have one common feature: they can be subjected to biodegradation using different microbial compositions. Microbial decomposition of wastes as meets the main crite-ria of safety and power consumption. We investigated the effectiveness of method of disposed personal hygiene means biodegradation by anaerobic thermophiles with further purification of obtained decomposition products from chemical solvents with the help of mesophilic isolates in microaerophile conditions. Bacteria of Clostridium genera were selected for cellulolysis be-cause of their high specific endoglucanasic activity which less depends on substrate nature and relatively high growth rate on cellulose contaning substrates. As result some strains in case of optimal conditions (substrata pretreating, pH correction) decomposed means of personal hygiene with level of biodegradation up to 90With the purpose of purification, liqiud medi-ums originating from Closrtidium sp. exhibiting used like substrates for cellololitic fungi. It was shown that the cultures are able to change pH of media from slow-acid to neutral. Also the effectiveness of plant wastes biodegradation (vegetables homogenates) was studied using associations of mesophile aerobes trophically adapted to substrates. Rate of biodestruction of dry mass varied near 76To purify liquid products of biodegradation from chemicals cellulolytic fungal strains as well as bacterial mesophylic association was used. Prevalence of cultures for purification was depended on pH of culture liquors. Chemical content of gaseous phase of cul-ture liquors was also studied. As it comes from chromatomass spectrometry data there was tremendous decrease of organic admixtures in liquid products of biodegradation after purifi-cation by fungal and bacterial cultures. These cultures were capable to support sustainable growth, feeding by metabolites of bacteria, which perform primary biodegradation. Also there was evaluated prospective of application of biofuel cells in the process of biotransformation of different substrates. Application of electrogenic bacteria could be perspective approach in wastes biodegradation technology.

  18. The High-Throughput Protein Sample Production Platform of the Northeast Structural Genomics Consortium

    PubMed Central

    Xiao, Rong; Anderson, Stephen; Aramini, James; Belote, Rachel; Buchwald, William A.; Ciccosanti, Colleen; Conover, Ken; Everett, John K.; Hamilton, Keith; Huang, Yuanpeng Janet; Janjua, Haleema; Jiang, Mei; Kornhaber, Gregory J.; Lee, Dong Yup; Locke, Jessica Y.; Ma, Li-Chung; Maglaqui, Melissa; Mao, Lei; Mitra, Saheli; Patel, Dayaban; Rossi, Paolo; Sahdev, Seema; Sharma, Seema; Shastry, Ritu; Swapna, G.V.T.; Tong, Saichu N.; Wang, Dongyan; Wang, Huang; Zhao, Li; Montelione, Gaetano T.; Acton, Thomas B.

    2014-01-01

    We describe the core Protein Production Platform of the Northeast Structural Genomics Consortium (NESG) and outline the strategies used for producing high-quality protein samples. The platform is centered on the cloning, expression and purification of 6X-His-tagged proteins using T7-based Escherichia coli systems. The 6X-His tag allows for similar purification procedures for most targets and implementation of high-throughput (HTP) parallel methods. In most cases, the 6X-His-tagged proteins are sufficiently purified (> 97% homogeneity) using a HTP two-step purification protocol for most structural studies. Using this platform, the open reading frames of over 16,000 different targeted proteins (or domains) have been cloned as > 26,000 constructs. Over the past nine years, more than 16,000 of these expressed protein, and more than 4,400 proteins (or domains) have been purified to homogeneity in tens of milligram quantities (see Summary Statistics, http://nesg.org/statistics.html). Using these samples, the NESG has deposited more than 900 new protein structures to the Protein Data Bank (PDB). The methods described here are effective in producing eukaryotic and prokaryotic protein samples in E. coli. This paper summarizes some of the updates made to the protein production pipeline in the last five years, corresponding to phase 2 of the NIGMS Protein Structure Initiative (PSI-2) project. The NESG Protein Production Platform is suitable for implementation in a large individual laboratory or by a small group of collaborating investigators. These advanced automated and/or parallel cloning, expression, purification, and biophysical screening technologies are of broad value to the structural biology, functional proteomics, and structural genomics communities. PMID:20688167

  19. Characterization of Molecules Binding to the 70K N-terminal Region of Fibronectin by IFAST Purification Coupled with Mass Spectrometry

    PubMed Central

    Moussavi-Harami, S. Farshid; Annis, Douglas S.; Ma, Wenjiang; Berry, Scott M.; Coughlin, Emma E.; Strotman, Lindsay N.; Maurer, Lisa M.; Westphall, Michael S.; Coon, Joshua J.; Mosher, Deane F.; Beebe, David J.

    2013-01-01

    Fibronectin (Fn) is a large glycoprotein present in plasma and extracellular matrix and is important for many processes. Within Fn the 70kDa N-terminal region (70k-Fn) is involved in cell-mediated Fn assembly, a process that contributes to embryogenesis, development, and platelet thrombus formation. In addition, major human pathogens including Staphlycoccus aureus and Streptococcus pyogenes, bind the 70k-Fn region by a novel form of protein-protein interaction called β-zipper formation, facilitating bacterial spread and colonization. Knowledge of blood plasma and platelet proteins that interact with 70k-Fn by β-zipper formation is incomplete. In the current study, we aimed to characterize these proteins through affinity purification. For this affinity purification, we used a novel purification technique termed immiscible filtration assisted by surface tension (IFAST). The foundation of this technology is immiscible phase filtration, using a magnet to draw paramagnetic particle (PMP)-bound analyte through an immiscible barrier (oil or organic solvent) that separates an aqueous sample from an aqueous eluting buffer. The immiscible barrier functions to remove unbound proteins via exclusion rather than dilutive washing used in traditional isolation methods. We identified 31 interactors from plasma, of which only seven were previously known to interact with Fn. Furthermore, five proteins were identified to interact with 70k-Fn from platelet lysate, of which one was previously known. These results demonstrate that IFAST offers advantages for proteomic studies of interacting molecules in that the technique requires small sample volumes, can be done with high enough throughput to sample multiple interaction conditions, and is amenable to exploratory mass spectrometric and confirmatory immuno-blotting read-outs. PMID:23750785

  20. Effect of pulsed corona discharge voltage and feed gas flow rate on dissolved ozone concentration

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Prasetyaningrum, A., E-mail: ajiprasetyaningrum@gmail.com; Ratnawati,; Jos, B.

    Ozonization is one of the methods extensively used for water purification and degradation of organic materials. Ozone (O{sub 3}) is recognized as a powerful oxidizing agent. Due to its strong oxidability and better environmental friendless, ozone increasing being used in domestic and industrial applications. Current technology in ozone production utilizes several techniques (corona discharge, ultra violet radiation and electrolysis). This experiment aimed to evaluating effect of voltage and gas flow rate on ozone production with corona discharge. The system consists of two net-type stainless steel electrode placed in a dielectric barrier. Three pulsed voltage (20, 30, 40 KV) and flowmore » rate (5, 10, 15 L/min) were prepare for operation variable at high frequency (3.7 kHz) with AC pulsed power supply. The dissolved ozone concentration depends on the applied high-voltage level, gas flow rate and the discharge exposure duration. The ozone concentration increases with decreasing gas flow rate. Dissolved ozone concentrations greater than 200 ppm can be obtained with a minimum voltage 40 kV.« less

  1. Cotton-based Cellulose Nanomaterials for Applications in Composites and Electronics

    NASA Astrophysics Data System (ADS)

    Farahbakhsh, Nasim

    A modern society demands development of highly valued and sustainable products via innovative process technologies and utilizing bio-based alternatives for petroleum based materials. Systematic comparative study of nanocellulose particles as a biodegradable and renewable reinforcing agent can help to develop criteria for selecting an appropriate candidate to be incorporated in polymer nanocomposites. Of particular interest has been nanocellulosic materials including cellulose nanocrystal (CNC) and micro/nanofibrilated cellulose (MFC/NFC) which possess a hierarchical structure that permits an ordered structure with unique properties that has served as building blocks for the design of green and novel materials composites for applications in flexible electronics, medicine and composites. Key differences exist in nanocellulosic materials as a result the process by which the material is produced. This research demonstrates the applicability for the use of recycled cotton as promising sustainable material to be utilized as a substrate for electronic application and a reinforcing agent choice that can be produced without any intensive purification process and be applied to synthetic-based polymer nanocomposites in melt-processing. (Abstract shortened by ProQuest.).

  2. Pooled protein immunization for identification of cell surface antigens in Streptococcus sanguinis.

    PubMed

    Ge, Xiuchun; Kitten, Todd; Munro, Cindy L; Conrad, Daniel H; Xu, Ping

    2010-07-26

    Available bacterial genomes provide opportunities for screening vaccines by reverse vaccinology. Efficient identification of surface antigens is required to reduce time and animal cost in this technology. We developed an approach to identify surface antigens rapidly in Streptococcus sanguinis, a common infective endocarditis causative species. We applied bioinformatics for antigen prediction and pooled antigens for immunization. Forty-seven surface-exposed proteins including 28 lipoproteins and 19 cell wall-anchored proteins were chosen based on computer algorithms and comparative genomic analyses. Eight proteins among these candidates and 2 other proteins were pooled together to immunize rabbits. The antiserum reacted strongly with each protein and with S. sanguinis whole cells. Affinity chromatography was used to purify the antibodies to 9 of the antigen pool components. Competitive ELISA and FACS results indicated that these 9 proteins were exposed on S. sanguinis cell surfaces. The purified antibodies had demonstrable opsonic activity. The results indicate that immunization with pooled proteins, in combination with affinity purification, and comprehensive immunological assays may facilitate cell surface antigen identification to combat infectious diseases.

  3. Pooled Protein Immunization for Identification of Cell Surface Antigens in Streptococcus sanguinis

    PubMed Central

    Ge, Xiuchun; Kitten, Todd; Munro, Cindy L.; Conrad, Daniel H.; Xu, Ping

    2010-01-01

    Background Available bacterial genomes provide opportunities for screening vaccines by reverse vaccinology. Efficient identification of surface antigens is required to reduce time and animal cost in this technology. We developed an approach to identify surface antigens rapidly in Streptococcus sanguinis, a common infective endocarditis causative species. Methods and Findings We applied bioinformatics for antigen prediction and pooled antigens for immunization. Forty-seven surface-exposed proteins including 28 lipoproteins and 19 cell wall-anchored proteins were chosen based on computer algorithms and comparative genomic analyses. Eight proteins among these candidates and 2 other proteins were pooled together to immunize rabbits. The antiserum reacted strongly with each protein and with S. sanguinis whole cells. Affinity chromatography was used to purify the antibodies to 9 of the antigen pool components. Competitive ELISA and FACS results indicated that these 9 proteins were exposed on S. sanguinis cell surfaces. The purified antibodies had demonstrable opsonic activity. Conclusions The results indicate that immunization with pooled proteins, in combination with affinity purification, and comprehensive immunological assays may facilitate cell surface antigen identification to combat infectious diseases. PMID:20668678

  4. Effect of pulsed corona discharge voltage and feed gas flow rate on dissolved ozone concentration

    NASA Astrophysics Data System (ADS)

    Prasetyaningrum, A.; Ratnawati, Jos, B.

    2015-12-01

    Ozonization is one of the methods extensively used for water purification and degradation of organic materials. Ozone (O3) is recognized as a powerful oxidizing agent. Due to its strong oxidability and better environmental friendless, ozone increasing being used in domestic and industrial applications. Current technology in ozone production utilizes several techniques (corona discharge, ultra violet radiation and electrolysis). This experiment aimed to evaluating effect of voltage and gas flow rate on ozone production with corona discharge. The system consists of two net-type stainless steel electrode placed in a dielectric barrier. Three pulsed voltage (20, 30, 40 KV) and flow rate (5, 10, 15 L/min) were prepare for operation variable at high frequency (3.7 kHz) with AC pulsed power supply. The dissolved ozone concentration depends on the applied high-voltage level, gas flow rate and the discharge exposure duration. The ozone concentration increases with decreasing gas flow rate. Dissolved ozone concentrations greater than 200 ppm can be obtained with a minimum voltage 40 kV.

  5. Market Analysis and Material Evaluation of Coagulants for Reverse Osmosis Water Purification Units

    DTIC Science & Technology

    1990-09-28

    three identified primary coagulants. Electrocoagulation , an emerging technology, was also discovered as being a relatively new area of research with great...there is yet another avenue the Army needs to explore -- electrocoagulation . 4.3.2 Electrocoagulation | * Electrocoagulation processes utilize an...7). A typical electrocoagulator consists of a combination of a treating chamber, which operates as an electrolytic cell, with a water-inlet and a

  6. Perspective of Membrane Technology in Dairy Industry: A Review

    PubMed Central

    Kumar, Pavan; Sharma, Neelesh; Ranjan, Rajeev; Kumar, Sunil; Bhat, Z. F.; Jeong, Dong Kee

    2013-01-01

    Membrane technology has revolutionized the dairy sector. Different types of membranes are used in the industry for various purposes like extending the shelf life of milk without exposure to heat treatment, standardization of the major components of milk for tailoring new products as well increasing yield and quality of the dairy products, and concentrating, fractionation and purification of milk components especially valuable milk proteins in their natural state. In the cheese industry, membranes increase the yield and quality of cheese and control the whey volume, by concentrating the cheese milk. With the advancement of newer technology in membrane processes, it is possible to recover growth factor from whey. With the introduction of superior quality membranes as well as newer technology, the major limitation of membranes, fouling or blockage has been overcome to a greater extent. PMID:25049918

  7. Perspective of membrane technology in dairy industry: a review.

    PubMed

    Kumar, Pavan; Sharma, Neelesh; Ranjan, Rajeev; Kumar, Sunil; Bhat, Z F; Jeong, Dong Kee

    2013-09-01

    Membrane technology has revolutionized the dairy sector. Different types of membranes are used in the industry for various purposes like extending the shelf life of milk without exposure to heat treatment, standardization of the major components of milk for tailoring new products as well increasing yield and quality of the dairy products, and concentrating, fractionation and purification of milk components especially valuable milk proteins in their natural state. In the cheese industry, membranes increase the yield and quality of cheese and control the whey volume, by concentrating the cheese milk. With the advancement of newer technology in membrane processes, it is possible to recover growth factor from whey. With the introduction of superior quality membranes as well as newer technology, the major limitation of membranes, fouling or blockage has been overcome to a greater extent.

  8. Second International Conference on Accelerating Biopharmaceutical Development

    PubMed Central

    2009-01-01

    The Second International Conference on Accelerating Biopharmaceutical Development was held in Coronado, California. The meeting was organized by the Society for Biological Engineering (SBE) and the American Institute of Chemical Engineers (AIChE); SBE is a technological community of the AIChE. Bob Adamson (Wyeth) and Chuck Goochee (Centocor) were co-chairs of the event, which had the theme “Delivering cost-effective, robust processes and methods quickly and efficiently.” The first day focused on emerging disruptive technologies and cutting-edge analytical techniques. Day two featured presentations on accelerated cell culture process development, critical quality attributes, specifications and comparability, and high throughput protein formulation development. The final day was dedicated to discussion of technology options and new analysis methods provided by emerging disruptive technologies; functional interaction, integration and synergy in platform development; and rapid and economic purification process development. PMID:20065637

  9. Cork boiling wastewater treatment and reuse through combination of advanced oxidation technologies.

    PubMed

    Ponce-Robles, L; Miralles-Cuevas, S; Oller, I; Agüera, A; Trinidad-Lozano, M J; Yuste, F J; Malato, S

    2017-03-01

    Industrial preparation of cork consists of its immersion for approximately 1 hour in boiling water. The use of herbicides and pesticides in oak tree forests leads to absorption of these compounds by cork; thus, after boiling process, they are present in wastewater. Cork boiling wastewater shows low biodegradability and high acute toxicity involving partial inhibition of their biodegradation when conventional biological treatment is applied. In this work, a treatment line strategy based on the combination of advanced physicochemical technologies is proposed. The final objective is the reuse of wastewater in the cork boiling process; thus, reducing consumption of fresh water in the industrial process itself. Coagulation pre-treatment with 0.5 g/L of FeCl 3 attained the highest turbidity elimination (86 %) and 29 % of DOC elimination. Similar DOC removal was attained when using 1 g/L of ECOTAN BIO (selected for ozonation tests), accompanied of 64 % of turbidity removal. Ozonation treatments showed less efficiency in the complete oxidation of cork boiling wastewater, compared to solar photo-Fenton process, under the studied conditions. Nanofiltration system was successfully employed as a final purification step with the aim of obtaining a high-quality reusable permeate stream. Monitoring of unknown compounds by LC-QTOF-MS allowed the qualitative evaluation of the whole process. Acute and chronic toxicity as well as biodegradability assays were performed throughout the whole proposed treatment line.

  10. Sunlight, iron and radicals to tackle the resistant leftovers of biotreated winery wastewater.

    PubMed

    Ioannou, Lida; Velegraki, Theodora; Michael, Costas; Mantzavinos, Dionissios; Fatta-Kassinos, Despo

    2013-04-01

    Winery wastewater is characterized by high organic content consisting of alcohols, acids and recalcitrant high-molecular-weight compounds (e.g. polyphenols, tannins and lignins). So far, biological treatment constitutes the best available technology for such effluents that are characterized by high seasonal variability; however the strict legislation applied on the reclamation and reuse of wastewaters for irrigation purposes introduces the need for further treatment of the bioresistant fraction of winery effluents. In this context, the use of alternative treatment technologies, aiming to mineralize or transform refractory molecules into others which could be further biodegraded, is a matter of great concern. In this study, a winery effluent that had already been treated in a sequencing batch reactor was subjected to further purification by homogeneous and heterogeneous solar Fenton oxidation processes. The effect of various operating variables such as catalyst and oxidant concentration, initial pH, temperature and lamp power on the abatement of chemical oxygen demand (COD), dissolved organic carbon (DOC), color, total phenolics and ecotoxicity has been assessed in the homogeneous solar Fenton process. In addition, a comparative assessment between homogeneous and heterogeneous solar Fenton processes was performed. In the present study the homogeneous solar Fenton process has been demonstrated to be the most effective process, yielding COD, DOC and total phenolics removal of about 69%, 48% and 71% in 120 min of the photocatalytic treatment, respectively.

  11. Industrial enzymatic production of cephalosporin-based beta-lactams.

    PubMed

    Barber, Michael S; Giesecke, Ulrich; Reichert, Arno; Minas, Wolfgang

    2004-01-01

    Cephalosporins are chemically closely related to penicillins both work by inhibiting the cell wall synthesis of bacteria. The first generation cephalosporins entered the market in 1964. Second and third generation cephalosporins were subsequently developed that were more powerful than the original products. Fourth generation cephalosporins are now reaching the market. Each newer generation of cephalosporins has greater Gram-negative antimicrobial properties than the preceding generation. Conversely, the 'older' generations of cephalosporins have greater Gram-positive (Staphylococcus and Streptococcus) coverage than the 'newer' generations. Frequency of dosing decreases and palatability generally improve with increasing generations. The advent of fourth generation cephalosporins with the launch of cefepime extended the spectrum against Gram-positive organisms without a significant loss of activity towards Gram-negative bacteria. Its greater stability to beta-lactamases increases its efficacy against drug-resistant bacteria. In this review we present the current situation of this mature market. In addition, we present the current state of the technologies employed for the production of cephalosporins, focusing on the new and environmentally safer 'green' routes to the products. Starting with the fermentation and purification of CPC, enzymatic conversion in conjunction with aqueous chemistry will lead to some key intermediates such as 7-ACA, TDA and TTA, which then can be converted into the active pharmaceutical ingredient (API), again applying biocatalytic technologies and aqueous chemistry. Examples for the costing of selected products are provided as well.

  12. Applying high-throughput methods to develop a purification process for a highly glycosylated protein.

    PubMed

    Sanaie, Nooshafarin; Cecchini, Douglas; Pieracci, John

    2012-10-01

    Micro-scale chromatography formats are becoming more routinely used in purification process development because of their ability to rapidly screen large number of process conditions at a time with minimal material. Given the usual constraints that exist on development timelines and resources, these systems can provide a means to maximize process knowledge and process robustness compared to traditional packed column formats. In this work, a high-throughput, 96-well filter plate format was used in the development of the cation exchange and hydrophobic interaction chromatography steps of a purification process designed to alter the glycoform distribution of a small protein. The significant input parameters affecting process performance were rapidly identified for both steps and preliminary operating conditions were identified. These ranges were verified in a packed chromatography column in order to assess the ability of the 96-well plate to predict packed column performance. In both steps, the 96-well plate format consistently led to underestimated glycoform-enrichment levels and to overestimated product recovery rates compared to the column-based approach. These studies demonstrate that the plate format can be used as a screening tool to narrow the operating ranges prior to further optimization on packed chromatography columns. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Efficient synthesis of pure monotosylated beta-cyclodextrin and its dimers.

    PubMed

    Tripodo, Giuseppe; Wischke, Christian; Neffe, Axel T; Lendlein, Andreas

    2013-11-15

    6-O-Monotosyl-β-cyclodextrin (mono-Ts-βCD) is one of the most important intermediates in the production of substituted βCD. So far, performing the monotosylation reaction and, in particular, the purification steps was challenging, relied on toxic solvents, and resulted in long and expensive procedures at, importantly, low yields. Here, the reaction of cyclodextrin with p-toluenesulfonyl chloride in aqueous environment is described to obtain a highly pure mono-Ts-βCD, for which a single-step purification with a cation exchange resin was applied. With this synthetic route and purification, yields could be increased from typically <10-15% to 35%, and organic solvents could be avoided. As characterized by FTIR, mass spectrometry, elemental analysis, and NMR, mono-Ts-βCD was obtained with a molar purity of >98mol%. From mono-Ts-βCD, β-cyclodextrin dimers linked by ethylenediamine (bis-Et-βCD) were successfully prepared (yield 93%, purity 96mol%) in a one-step approach using an anion exchange resin to trap leaving groups that typically interfere in the reaction. This synthesis procedure with a direct collection of side-products may be a general strategy applicable for nucleophilic substitution of tosylated cyclodextrins. Copyright © 2013 Elsevier Ltd. All rights reserved.

  14. The concurrent growth of plants and chemical purification of wastewater used as a hydroponic unit.

    PubMed

    Jurdi, M; Soufi, M; Acra, A

    1987-01-01

    In this study the seedling of a variety of plants were successfully grown hydroponically on raw wastewater obtained from one of the main sewer outfalls in Beirut. In the first phase, a series of experiments was run on a batch system in glass or plastic containers provided with aeration facilities. A continuous-flow system with recirculation was adopted in the second phase. Iron supplementation was applied in all cases to compensate for its deficiency in the raw wastewater used. The immediate and ultimate objectives of the project were threefold: (a) to demonstrate the feasibility of utilizing as a hydroponic medium untreated municipal wastewater having relatively high mean values for BOD and mineral content; (b) to achieve the growth of useful plants on such readily available hydroponic media, thereby saving on fertilizers and scarce water resources; and (c) reclamation of the wastewater through biological purification leading to the gradual depletion of the nutritive constituents. Experimental conditions are described, and the data presented leads to the conclusion that the system is practicable on a laboratory scale. It has great potential for trial on a pilot scale prior to field applications in developing countries suffering from water shortage and hard currency expended on imported fertilizers and wastewater purification facilities.

  15. Extraction and purification of capsaicin from capsicum oleoresin using an aqueous two-phase system combined with chromatography.

    PubMed

    Fan, Yong; Lu, Yan-Min; Yu, Bin; Tan, Cong-Ping; Cui, Bo

    2017-09-15

    Capsaicin was extracted from capsicum oleoresin using an aqueous two-phase system (ATPS) composed of an ethylene oxide-propylene oxide (EOPO) copolymer, salt and ethanol. Capsaicin was concentrated in the top polymer-rich phase. To determine the optimal conditions, the partitioning of capsaicin in the ATPS was investigated, considering a single-factor experiment including the salt concentration, polymer concentration, buffer pH, ethanol concentration, sample loading and extraction duration. Response surface methodology was applied to investigate the effects of the polymer concentration, buffer pH and sample loading on capsaicin partitioning. A capsaicin yield of 95.5% was obtained using the optimal extraction system, which consisted of 16.3% UCON 50-HB-5100/10% K 2 HPO 4 /1% ethanol, a buffer pH of 4.35 and 0.24g of capsicum oleoresin. Capsaicin was purified from the capsaicinoid extract using a two-step macroporous adsorption resin (MAR) method. After purification using non-polar MAR ADS-17, the recovery and purity of capsaicin were 83.7% and 50.3%, respectively. After purification using weakly polar MAR AB-8, the recovery and purity of capsaicin were 88.0% and 85.1%, respectively. Copyright © 2017. Published by Elsevier B.V.

  16. Purification of gamma-amino butyric acid (GABA) from fermentation of defatted rice bran extract by using ion exchange resin

    NASA Astrophysics Data System (ADS)

    Tuan Nha, Vi; Phung, Le Thi Kim; Dat, Lai Quoc

    2017-09-01

    Rice bran is one of the significant byproducts of rice processing with 10 %w/w of constitution of whole rice grain. It is rich in nutrient compounds, including glutamic acid. Thus, it could be utilized for the fermentation with Lactobateria for synthesis of GABA, a valuable bioactive for antihypertensive effects. However, the concentration and purity of GABA in fermentation broth of defatted rice bran extract is low for production of GABA drug. This research focused on the purification of GABA from the fermentation broth of defatted rice bran extract by using cation exchange resin. The results indicate that, the adsorption isotherm of GABA by Purelite C100 showed the good agreement with Freundlich model, with high adsorption capacity. The effects of pH and concentration of NaCl in eluent on the elution were also investigated. The obtained results show that, at the operating conditions of elution as follows: pH 6.5, 0.8 M of NaCl in eluent, 0.43 of bed volume; concentration of GABA in accumulative eluent, the purity and recovery yield of GABA were 743.8 ppm, 44.0% and 84.2%, respectively. Results imply that, it is feasible to apply cation exchange resin for purification of GABA from fermentation broth of defatted rice bran extract.

  17. ACUTE DIALYSIS QUALITY INITIATIVE (ADQI) XIV SEPSIS PHENOTYPES AND TARGETS FOR BLOOD PURIFICATION IN SEPSIS: THE BOGOTÁ CONSENSUS.

    PubMed

    Kellum, John A; Gómez, Hernando; Gómez, Alonso; Murray, Patrick; Ronco, Claudio

    2016-03-01

    Despite widespread use, there is currently no consensus on how extracorporeal blood purification therapies should be applied or studied in patients with sepsis. One major obstacle has been the lack of clear descriptions of specific sepsis phenotypes tied to mechanisms that would permit the identification of molecular targets. Current evidence suggests that sepsis-related morbidity and mortality involve widely different clinical phenotypes that variably include mitochondrial dysfunction, abnormalities of vascular biology including endothelial dysfunction and coagulopathy, epithelial dysfunction, and immune suppression and dysregulation. While most cases of sepsis involve some element of all of these pathobiologic processes, the magnitude of each varies greatly from patient to patient in part as a result of the pathogen and in part related to host-specific factors. Thus, the purpose of the fourteenth international consensus conference of acute dialysis quality initiative was to develop consensus for a conceptual model of sepsis-induced organ failure that can be treated by extracorporeal blood purification and possibly also with drugs or other therapies. We assembled a group of experts from around the world and used a modified Delphi method to reach consensus. Specific findings and recommendations for future research are provided in the four accompanying papers.

  18. QuEChERS-based purification method coupled to ultrahigh performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) to determine six quaternary ammonium compounds (QACs) in dairy products.

    PubMed

    Xian, Yanping; Dong, Hao; Wu, Yuluan; Guo, Xindong; Hou, Xiangchang; Wang, Bin

    2016-12-01

    QuEChERS-based purification coupled with UPLC-MS/MS method, was developed for six quaternary ammonium compounds (QACs) determination in dairy products. Powder samples were firstly dispersed by water. Protein in liquid milk was precipitated and sample solution was extracted by acetonitrile. QuEChERS-based purification was used to purify the solution. QACs were finally separated by HILIC column and detected in MRM mode of MS/MS under ESI(+). The stable isotope benzyl-2,3,4,5,6-d5-dimethyltetradecylammonium bromide (C14-BAC-d5) was used as an internal standard. This method was validated in terms of linearity, sensitivity, precision, accuracy. Linear relations were favorable for QACs over the selected concentration ranges of 0.2-50μg/L, with correlation coefficients greater than 0.999. The limits of detection (LODs) were in the range of 0.4-14.5μg/kg. Recoveries were between 91.2% and 115% with RSDs of 2.8-7.5% for intra-day precision and 3.7-6.7% for inter-day precision. This validated method was successfully applied to determine the QACs concentrations in dairy products. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Enzymatic Purification of Microplastics in Environmental Samples.

    PubMed

    Löder, Martin G J; Imhof, Hannes K; Ladehoff, Maike; Löschel, Lena A; Lorenz, Claudia; Mintenig, Svenja; Piehl, Sarah; Primpke, Sebastian; Schrank, Isabella; Laforsch, Christian; Gerdts, Gunnar

    2017-12-19

    Micro-Fourier transform infrared (micro-FTIR) spectroscopy and Raman spectroscopy enable the reliable identification and quantification of microplastics (MPs) in the lower micron range. Since concentrations of MPs in the environment are usually low, the large sample volumes required for these techniques lead to an excess of coenriched organic or inorganic materials. While inorganic materials can be separated from MPs using density separation, the organic fraction impedes the ability to conduct reliable analyses. Hence, the purification of MPs from organic materials is crucial prior to conducting an identification via spectroscopic techniques. Strong acidic or alkaline treatments bear the danger of degrading sensitive synthetic polymers. We suggest an alternative method, which uses a series of technical grade enzymes for purifying MPs in environmental samples. A basic enzymatic purification protocol (BEPP) proved to be efficient while reducing 98.3 ± 0.1% of the sample matrix in surface water samples. After showing a high recovery rate (84.5 ± 3.3%), the BEPP was successfully applied to environmental samples from the North Sea where numbers of MPs range from 0.05 to 4.42 items m -3 . Experiences with different environmental sample matrices were considered in an improved and universally applicable version of the BEPP, which is suitable for focal plane array detector (FPA)-based micro-FTIR analyses of water, wastewater, sediment, biota, and food samples.

  20. Production, Purification, and Identification of Cholest-4-en-3-one Produced by Cholesterol Oxidase from Rhodococcus sp. in Aqueous/Organic Biphasic System

    PubMed Central

    Wu, Ke; Li, Wei; Song, Jianrui; Li, Tao

    2015-01-01

    Cholest-4-en-3-one has positive uses against obesity, liver disease, and keratinization. It can be applied in the synthesis of steroid drugs as well. Most related studies are focused on preparation of cholest-4-en-3-one by using whole cells as catalysts, but production of high-quality cholest-4-en-3-one directly from cholesterol oxidase (COD) using an aqueous/organic two-phase system has been rarely explored. This study set up an enzymatic reaction system to produce cholest-4-en-3-one. We developed and optimized the enzymatic reaction system using COD from COX5-6 (a strain of Rhodococcus) instead of whole-cell biocatalyst. This not only simplifies and accelerates the production but also benefits the subsequent separation and purification process. Through extraction, washing, evaporation, column chromatography, and recrystallization, we got cholest-4-en-3-one with purity of 99.78%, which was identified by nuclear magnetic resonance, mass spectroscopy, and infrared spectroscopy. In addition, this optimized process of cholest-4-en-3-one production and purification can be easily scaled up for industrial production, which can largely decrease the cost and guarantee the purity of the product. PMID:25733914

  1. Air purification in industrial plants producing automotive rubber components in terms of energy efficiency

    NASA Astrophysics Data System (ADS)

    Grzebielec, Andrzej; Rusowicz, Artur; Szelągowski, Adam

    2017-04-01

    In automotive industry plants, which use injection molding machines for rubber processing, tar contaminates air to such an extent that air fails to enter standard heat recovery systems. Accumulated tar clogs ventilation heat recovery exchangers in just a few days. In the plant in which the research was conducted, tar contamination causes blockage of ventilation ducts. The effect of this phenomenon was that every half year channels had to be replaced with new ones, since the economic analysis has shown that cleaning them is not cost-efficient. Air temperature inside such plants is often, even in winter, higher than 30°C. The air, without any means of heat recovery, is discharged outside the buildings. The analyzed plant uses three types of media for production: hot water, cold water at 14°C (produced in a water chiller), and compressed air, generated in a unit with a rated power consumption of 180 kW. The aim of the study is to determine the energy efficiency improvement of this type of manufacturing plant. The main problem to solve is to provide an air purification process so that air can be used in heat recovery devices. The next problem to solve is to recover heat at such a temperature level that it would be possible to produce cold for technological purposes without air purification. Experimental studies have shown that air purification is feasible. By using one microjet head, a total of 75% of tar particles was removed from the air; by using 4 heads, a purification efficiency of 93% was obtained. This method of air purification causes air temperature to decrease from 35°C to 20°C, which significantly reduces the potential for heat recovery. The next step of the research was designing a cassette-plate heat exchanger to exchange heat without air purification. The economic analysis of such a solution revealed that replacing the heat exchanger with a new one even once a year was not cost-efficient. Another issue examined in the context of energy efficiency was the use of waste heat from the air compressor. Before any changes, the heat was picked up by a chilled water system. The idea was to use the heat for cold generation. Temperature of oil and air in the compressor exceeds 65°C, which makes it a perfect heat source for an adsorption refrigeration device. This solution reduced the cooling demand by 147 kW, thus reducing power consumption by 36.75 kW. This study shows that even in factories where air is heavily polluted with tar, there are huge potentials for energy recovery using existing technical solutions. It is important to note that problems of this kind should always be approached individually.

  2. Nonthermal plasma technology for organic destruction

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Heath, W.O.; Birmingham, J.G.

    1995-06-01

    Pacific Northwest Laboratory (PNL) is investigating the use of nonthermal, electrically driven plasmas for destroying organic contaminants near ambient temperatures and pressures. Three different plasma systems have been developed to treat organics in air, water, and soil. These systems are the Gas-Phase Corona Reactor (GPCR)III for treating air, the Liquid-Phase Corona Reactor for treating water, and In Situ Corona for treating soils. This presentation focuses on recent technical developments, commercial status, and project costs of OPCR as a cost-effective alternative to other air-purification technologies that are now in use to treat off-gases from site-remediation efforts as well as industrial emissions.

  3. Coagulant Recovery from Water Treatment Residuals: A Review of Applicable Technologies

    PubMed Central

    Keeley, J.; Jarvis, P.; Judd, S. J.

    2014-01-01

    Conventional water treatment consumes large quantities of coagulant and produces even greater volumes of sludge. Coagulant recovery (CR) presents an opportunity to reduce both the sludge quantities and the costs they incur, by regenerating and purifying coagulant before reuse. Recovery and purification must satisfy stringent potable regulations for harmful contaminants, while remaining competitive with commercial coagulants. These challenges have restricted uptake and lead research towards lower-gain, lower-risk alternatives. This review documents the context in which CR must be considered, before comparing the relative efficacies and bottlenecks of potential technologies, expediting identification of the major knowledge gaps and future research requirements. PMID:26064036

  4. New Trends and Technological Challenges in the Industrial Production and Purification of Fructo-oligosaccharides.

    PubMed

    Nobre, Clarisse; Teixeira, José A; Rodrigues, Lígia R

    2015-01-01

    An increased commercial interest in fructo-oligosaccharides (FOS) has emerged in the last decade due to their prebiotic activity. At large scale, the FOS are produced by microbial enzymes from sucrose. A mixture of FOS and other saccharides is obtained in this process. The presence of such saccharides reduces the prebiotic, caloric, and cariogenic value of the final product. Therefore, many efforts have been conducted to obtain a product with increased FOS purity. This review comprises the most important technological and physicochemical aspects including FOS production and recovery processes; safety, dose and health claims concerning its intake; and commercially available FOS.

  5. RNActive® Technology: Generation and Testing of Stable and Immunogenic mRNA Vaccines.

    PubMed

    Rauch, Susanne; Lutz, Johannes; Kowalczyk, Aleksandra; Schlake, Thomas; Heidenreich, Regina

    2017-01-01

    Developing effective mRNA vaccines poses certain challenges concerning mRNA stability and ability to induce sufficient immune stimulation and requires a specific panel of techniques for production and testing. Here, we describe the production of stabilized mRNA with enhanced immunogenicity, generated using conventional nucleotides only, by introducing changes to the mRNA sequence and by complexation with the nucleotide-binding peptide protamine (RNActive® technology). Methods described here include the synthesis, purification, and protamine complexation of mRNA vaccines as well as a comprehensive panel of in vitro and in vivo methods for evaluation of vaccine quality and immunogenicity.

  6. Parallel production and verification of protein products using a novel high-throughput screening method.

    PubMed

    Tegel, Hanna; Yderland, Louise; Boström, Tove; Eriksson, Cecilia; Ukkonen, Kaisa; Vasala, Antti; Neubauer, Peter; Ottosson, Jenny; Hober, Sophia

    2011-08-01

    Protein production and analysis in a parallel fashion is today applied in laboratories worldwide and there is a great need to improve the techniques and systems used for this purpose. In order to save time and money, a fast and reliable screening method for analysis of protein production and also verification of the protein product is desired. Here, a micro-scale protocol for the parallel production and screening of 96 proteins in plate format is described. Protein capture was achieved using immobilized metal affinity chromatography and the product was verified using matrix-assisted laser desorption ionization time-of-flight MS. In order to obtain sufficiently high cell densities and product yield in the small-volume cultivations, the EnBase® cultivation technology was applied, which enables cultivation in as small volumes as 150 μL. Here, the efficiency of the method is demonstrated by producing 96 human, recombinant proteins, both in micro-scale and using a standard full-scale protocol and comparing the results in regard to both protein identity and sample purity. The results obtained are highly comparable to those acquired through employing standard full-scale purification protocols, thus validating this method as a successful initial screening step before protein production at a larger scale. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Polyelectrolyte-coated ion exchangers for cell-resistant expanded bed adsorption.

    PubMed

    Dainiak, Maria B; Galaev, Igor Yu; Mattiasson, Bo

    2002-01-01

    Adsorption chromatography in expanded beds is a widely used technology for direct capture of target proteins from fermentation broths. However, in many cases this method cannot be applied as a result of the strong tendency of cells or cell debris to interact with the adsorbent beads. To prevent contamination of the expanded bed with the biomass, STREAMLINE DEAE, anion exchanger designed for expanded bed adsorption, was modified with a layer of poly(acrylic acid) (PAA). The shielding layer of polyelectrolyte was attached to the surface of the matrix beads via electrostatic interactions. PAA with a high degree of polymerization was chosen to prevent diffusion of large polymer molecules into the pores of adsorbent. Thus, the shielding layer of PAA was adsorbed only at the mouth of the pores of STREAMLINE DEAE beads and only marginally decreased the binding capacity of the ion exchanger for bovine serum albumin, the model protein in this study. PAA-coated STREAMLINE DEAE practically did not interact with yeast cells, which otherwise bound strongly to the native adsorbent at neutral conditions. Cell-resistant PAA-coated anion exchanger was successfully used for isolation of BSA from the model protein mixture containing BSA, lysozyme (positively charged at applied conditions), and yeast cells. The layer of PAA was stable under mild elution conditions, and the modified adsorbent could be used in the repeated purification cycles.

  8. Silver nanocluster catalytic microreactors for water purification

    NASA Astrophysics Data System (ADS)

    Da Silva, B.; Habibi, M.; Ognier, S.; Schelcher, G.; Mostafavi-Amjad, J.; Khalesifard, H. R. M.; Tatoulian, M.; Bonn, D.

    2016-07-01

    A new method for the elaboration of a novel type of catalytic microsystem with a high specific area catalyst is developed. A silver nanocluster catalytic microreactor was elaborated by doping a soda-lime glass with a silver salt. By applying a high power laser beam to the glass, silver nanoclusters are obtained at one of the surfaces which were characterized by BET measurements and AFM. A microfluidic chip was obtained by sealing the silver coated glass with a NOA 81 microchannel. The catalytic activity of the silver nanoclusters was then tested for the efficiency of water purification by using catalytic ozonation to oxidize an organic pollutant. The silver nanoclusters were found to be very stable in the microreactor and efficiently oxidized the pollutant, in spite of the very short residence times in the microchannel. This opens the way to study catalytic reactions in microchannels without the need of introducing the catalyst as a powder or manufacturing complex packed bed microreactors.

  9. PREPARATIVE ISOLATION AND PURIFICATION OF CHEMICAL CONSTITUENTS OF BELAMCANDA BY MPLC, HSCCC AND PREP-HPLC

    PubMed Central

    Wang, Xiaohong; Liang, Yong; Peng, Cuilin; Xie, Huichun; Pan, Man; Zhang, Tianyou; Ito, Yoichiro

    2010-01-01

    Combined with medium-pressure liquid chromatography (MPLC) and preparative high-pressure liquid chromatography (Prep-HPLC), high-speed countercurrent chromatography (HSCCC) was successfully applied for separation and purification of isoflavonoids from the extract of belamcanda. HSCCC separation was performed on a two-phase solvent system composed of methyl tert-butyl ether -ethyl acetate - n-butyl alcohol – acetonitrile −0.1% aqueous trifluoroacetic acid at a volume radio of 1:2:1:1:5. Semi-purified peak fractions from HSCCC separation were further purified by Prep-HPLC. Nine well-separated fractions were analyzed by HPLC-UV absorption spectrometry to determine their purities and characterized with ESI-MSn. Except for peaksland VII (unknown) seven compounds were identified as apocynin (peak II), mangiferin (peak III), 7-O-methylmangiferin (peak IV), hispidulin (peak V), 3′-hydroxyltectoridin (peak VI), iristectorin B (peak VII), isoiridin (peak IX). PMID:21552369

  10. Development of nitrocellulose membrane filters impregnated with different biosynthesized silver nanoparticles applied to water purification.

    PubMed

    Fernández, Jorge G; Almeida, César A; Fernández-Baldo, Martín A; Felici, Emiliano; Raba, Julio; Sanz, María I

    2016-01-01

    Bactericidal water filters were developed. For this purpose, nitrocellulose membrane filters were impregnated with different biosynthesized silver nanoparticles. Silver nanoparticles (AgNPs) from Aspergillus niger (AgNPs-Asp), Cryptococcus laurentii (AgNPs-Cry) and Rhodotorula glutinis (AgNPs-Rho) were used for impregnating nitrocellulose filters. The bactericidal properties of these nanoparticles against Escherichia coli, Enterococcus faecalis and Pseudomona aeruginosa were successfully demonstrated. The higher antimicrobial effect was observed for AgNPs-Rho. This fact would be related not only to the smallest particles, but also to polysaccharides groups that surrounding these particles. Moreover, in this study, complete inhibition of bacterial growth was observed on nitrocellulose membrane filters impregnated with 1 mg L(-1) of biosynthesized AgNPs. This concentration was able to reduce the bacteria colony count by over 5 orders of magnitude, doing suitable for a water purification device. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Toxoplasma gondii tissue cyst purification using Percoll gradients

    PubMed Central

    Watts, Elizabeth A.; Dhara, Animesh; Sinai, Anthony P.

    2017-01-01

    The protozoan parasite Toxoplasma gondii is capable of infecting all warm blooded animals and humans. Infectious, transmissible forms of the parasite include oocysts produced by the sexual cycle within the definitive feline host and tissue cysts that form Toxoplasma in the CNS and muscle during the asexual cycle within all chronically infected warm-blooded hosts. These tissue cysts are populated with slow growing bradyzoites which have been until recently thought to be dormant entities in the context of immune sufficiency. Reactivation to active growth during immune suppression is of critical clinical importance. Yet we know little about tissue cysts or the bradyzoites they house as the diversity of tissue cysts cannot be replicated in cell culture systems. Our optimization of tissue cyst purification from the brains of infected mice using Percoll gradients provides an efficient means to recover in vivo derived tissue cysts that can be applied to imaging, cell-biologic, biochemical, transcriptomic and proteomic analyses. PMID:28510363

  12. Electrospun magnetically separable calcium ferrite nanofibers for photocatalytic water purification

    NASA Astrophysics Data System (ADS)

    EL-Rafei, A. M.; El-Kalliny, Amer S.; Gad-Allah, Tarek A.

    2017-04-01

    Three-dimensional random calcium ferrite, CaFe2O4, nanofibers (NFs) were successfully prepared via the electrospinning method. The effect of calcination temperature on the characteristics of the as-spun NFs was investigated. X-ray diffraction analysis showed that CaFe2O4 phase crystallized as a main phase at 700 °C and as a sole phase at 1000 °C. Field emission scanning electron microscopy emphasized that CaFe2O4 NFs were fabricated with diameters in the range of 50-150 nm and each fiber was composed of 20-50 nm grains. Magnetic hysteresis loops revealed superparamagnetic behavior for the prepared NFs. These NFs produced active hydroxyl radicals under simulated solar light irradiation making them recommendable for photocatalysis applications in water purification. In the meantime, these NFs can be easily separated from the treated water by applying an external magnetic field.

  13. Chemical looping integration with a carbon dioxide gas purification unit

    DOEpatents

    Andrus, Jr., Herbert E.; Jukkola, Glen D.; Thibeault, Paul R.; Liljedahl, Gregory N.

    2017-01-24

    A chemical looping system that contains an oxidizer and a reducer is in fluid communication with a gas purification unit. The gas purification unit has at least one compressor, at least one dryer; and at least one distillation purification system; where the gas purification unit is operative to separate carbon dioxide from other contaminants present in the flue gas stream; and where the gas purification unit is operative to recycle the contaminants to the chemical looping system in the form of a vent gas that provides lift for reactants in the reducer.

  14. The Reserve Forces Impact on Theater Security Cooperation

    DTIC Science & Technology

    2013-03-01

    in South America there are needs for purification is spoiled water, prophylactics, and typhoid , malaria, and yellow fever vaccines. Professions...Theater Security Cooperation (TSC) is a Phase 0 ( shaping ) instrument of regional commanders and is applied with the intent of precluding the use of...military force. The U.S. Army War College (USAWC) Theater Campaign Planning Handbook describes Phase 0 operations as “those shaping activities

  15. Scalable, cGMP-compatible purification of extracellular vesicles carrying bioactive human heterodimeric IL-15/lactadherin complexes

    PubMed Central

    Watson, Dionysios C.; Yung, Bryant C.; Bergamaschi, Cristina; Chowdhury, Bhabadeb; Bear, Jenifer; Stellas, Dimitris; Morales-Kastresana, Aizea; Jones, Jennifer C.; Felber, Barbara K.; Chen, Xiaoyuan; Pavlakis, George N.

    2018-01-01

    ABSTRACT The development of extracellular vesicles (EV) for therapeutic applications is contingent upon the establishment of reproducible, scalable, and high-throughput methods for the production and purification of clinical grade EV. Methods including ultracentrifugation (U/C), ultrafiltration, immunoprecipitation, and size-exclusion chromatography (SEC) have been employed to isolate EV, each facing limitations such as efficiency, particle purity, lengthy processing time, and/or sample volume. We developed a cGMP-compatible method for the scalable production, concentration, and isolation of EV through a strategy involving bioreactor culture, tangential flow filtration (TFF), and preparative SEC. We applied this purification method for the isolation of engineered EV carrying multiple complexes of a novel human immunostimulatory cytokine-fusion protein, heterodimeric IL-15 (hetIL-15)/lactadherin. HEK293 cells stably expressing the fusion cytokine were cultured in a hollow-fibre bioreactor. Conditioned medium was collected and EV were isolated comparing three procedures: U/C, SEC, or TFF + SEC. SEC demonstrated comparable particle recovery, size distribution, and hetIL-15 density as U/C purification. Relative to U/C, SEC preparations achieved a 100-fold reduction in ferritin concentration, a major protein-complex contaminant. Comparative proteomics suggested that SEC additionally decreased the abundance of cytoplasmic proteins not associated with EV. Combination of TFF and SEC allowed for bulk processing of large starting volumes, and resulted in bioactive EV, without significant loss in particle yield or changes in size, morphology, and hetIL-15/lactadherin density. Taken together, the combination of bioreactor culture with TFF + SEC comprises a scalable, efficient method for the production of highly purified, bioactive EV carrying hetIL-15/lactadherin, which may be useful in targeted cancer immunotherapy approaches. PMID:29535850

  16. Effects of antibody disulfide bond reduction on purification process performance and final drug substance stability

    PubMed Central

    Russell, Brian; Yang, Yanhong; Handlogten, Michael; Hudak, Suzanne; Cao, Mingyan; Wang, Jihong; Robbins, David; Ahuja, Sanjeev; Zhu, Min

    2017-01-01

    ABSTRACT Antibody disulfide bond reduction during monoclonal antibody (mAb) production is a phenomenon that has been attributed to the reducing enzymes from CHO cells acting on the mAb during the harvest process. However, the impact of antibody reduction on the downstream purification process has not been studied. During the production of an IgG2 mAb, antibody reduction was observed in the harvested cell culture fluid (HCCF), resulting in high fragment levels. In addition, aggregate levels increased during the low pH treatment step in the purification process. A correlation between the level of free thiol in the HCCF (as a result of antibody reduction) and aggregation during the low pH step was established, wherein higher levels of free thiol in the starting sample resulted in increased levels of aggregates during low pH treatment. The elevated levels of free thiol were not reduced over the course of purification, resulting in carry‐over of high free thiol content into the formulated drug substance. When the drug substance with high free thiols was monitored for product degradation at room temperature and 2–8°C, faster rates of aggregation were observed compared to the drug substance generated from HCCF that was purified immediately after harvest. Further, when antibody reduction mitigations (e.g., chilling, aeration, and addition of cystine) were applied, HCCF could be held for an extended period of time while providing the same product quality/stability as material that had been purified immediately after harvest. Biotechnol. Bioeng. 2017;114: 1264–1274. © 2017 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals Inc. PMID:28186329

  17. Continuous processing of recombinant proteins: integration of refolding and purification using simulated moving bed size-exclusion chromatography with buffer recycling.

    PubMed

    Wellhoefer, Martin; Sprinzl, Wolfgang; Hahn, Rainer; Jungbauer, Alois

    2014-04-11

    Continuous processing of recombinant proteins was accomplished by combining continuous matrix-assisted refolding and purification by tandem simulated moving bed (SMB) size-exclusion chromatography (SEC). Recombinant proteins, N(pro) fusion proteins from inclusion bodies were dissolved with NaOH and refolded in the SMB system with a closed-loop set-up with refolding buffer as the desorbent buffer and buffer recycling of the refolding buffer of the raffinate by tangential flow filtration. For further purification of the refolded proteins, a second SMB operation also based on SEC was added. The whole system could be operated isocratically with refolding buffer as the desorbent buffer, and buffer recycling could also be applied in the purification step. Thus, a significant reduction in buffer consumption was achieved. The system was evaluated with two proteins, the N(pro) fusion pep6His and N(pro) fusion MCP-1. Refolding solution, which contained residual N(pro) fusion peptide, the cleaved autoprotease N(pro), and the cleaved target peptide was used as feed solution. Full separation of the cleaved target peptide from residual proteins was achieved at a purity and recovery in the raffinate and extract, respectively, of approximately 100%. In addition, more than 99% of the refolding buffer of the raffinate was recycled. A comparison of throughput, productivity, and buffer consumption of the integrated continuous process with two batch processes demonstrated that up to 60-fold higher throughput, up to 180-fold higher productivity, and at least 28-fold lower buffer consumption can be obtained by the integrated continuous process, which compensates for the higher complexity. Copyright © 2014 Elsevier B.V. All rights reserved.

  18. New process for purifying high purity α1-antitrypsin from Cohn Fraction IV by chromatography: A promising method for the better utilization of plasma.

    PubMed

    Huangfu, Chaoji; Zhang, Jinchao; Ma, Yuyuan; Jia, Junting; Lv, Maomin; Zhao, Xiong; Zhang, Jingang

    2017-03-01

    α1-antitrypsin (AAT) is a 52kDa serine protease inhibitor that is abundant in plasma. It is synthesized mainly by hepatic cells, and widely used to treat patients with emphysema due to congenital deficiency of AAT. A new isolation method for the purification of AAT from Cohn Fraction IV (Cohn F IV) is described. Cohn F IV is usually discarded as a byproduct from Cohn process. Using Cohn F IV as starting material does not interfere with the production of other plasma proteins and the cost of purification could be reduced greatly. Parameters of each step during purification were optimized, 15% polyethyleneglycol (PEG) concentration and pH 5.2 for PEG precipitation, elution with 0.05M sodium acetate and pH 4.7 for ion-exchange chromatography, and two steps blue sepharose affinity chromatography were chosen for AAT purification. The final protein with purity of 98.17%, specific activity of 3893.29 IU/mg, and yield of 28.35%, was achieved. Western blotting was applied for qualitative identification of final product, which specifically reacted with goat anti-human AAT antibody. LC-ESI-MS/MS was also employed to confirm the final protein. High performance liquid chromatography was used to analyze the composition of purified protein suggesting that pure protein was achieved. The molecular weight of AAT is 51062.77Da which was identified by LC-MS-MS. The manufacturing process described here may make better use of human plasma with Cohn F IV as starting material. The simple process described in this study is simple and inexpensive, it has a potential value for large scale production. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Electron-driven and thermal chemistry during water-assisted purification of platinum nanomaterials generated by electron beam induced deposition

    PubMed Central

    Warneke, Jonas; Kopyra, Janina

    2018-01-01

    Focused electron beam induced deposition (FEBID) is a versatile tool for the direct-write fabrication of nanostructures on surfaces. However, FEBID nanostructures are usually highly contaminated by carbon originating from the precursor used in the process. Recently, it was shown that platinum nanostructures produced by FEBID can be efficiently purified by electron irradiation in the presence of water. If such processes can be transferred to FEBID deposits produced from other carbon-containing precursors, a new general approach to the generation of pure metallic nanostructures could be implemented. Therefore this study aims to understand the chemical reactions that are fundamental to the water-assisted purification of platinum FEBID deposits generated from trimethyl(methylcyclopentadienyl)platinum(IV) (MeCpPtMe3). The experiments performed under ultrahigh vacuum conditions apply a combination of different desorption experiments coupled with mass spectrometry to analyse reaction products. Electron-stimulated desorption monitors species that leave the surface during electron exposure while post-irradiation thermal desorption spectrometry reveals products that evolve during subsequent thermal treatment. In addition, desorption of volatile products was also observed when a deposit produced by electron exposure was subsequently brought into contact with water. The results distinguish between contributions of thermal chemistry, direct chemistry between water and the deposit, and electron-induced reactions that all contribute to the purification process. We discuss reaction kinetics for the main volatile products CO and CH4 to obtain mechanistic information. The results provide novel insights into the chemistry that occurs during purification of FEBID nanostructures with implications also for the stability of the carbonaceous matrix of nanogranular FEBID materials under humid conditions. PMID:29441253

  20. [Studying on purification technology of Resina Draconis phenol extracts based on design space method].

    PubMed

    Zhang, Jian; Zhang, Xin; Bi, Yu-An; Xu, Gui-Hong; Huang, Wen-Zhe; Wang, Zhen-Zhong; Xiao, Wei

    2017-09-01

    The "design space" method was used to optimize the purification process of Resina Draconis phenol extracts by using the concept of "quality derived from design" (QbD). The content and transfer rate of laurin B and 7,4'-dihydroxyflavone and yield of extract were selected as the critical quality attributes (CQA). Plackett-Burman design showed that the critical process parameters (CPP) were concentration of alkali, the amount of alkali and the temperature of alkali dissolution. Then the Box-Behnken design was used to establish the mathematical model between CQA and CPP. The variance analysis results showed that the P values of the five models were less than 0.05 and the mismatch values were all greater than 0.05, indicating that the model could well describe the relationship between CQA and CPP. Finally, the control limits of the above 5 indicators (content and transfer rate of laurine B and 7,4'-dihydroxyflavone, as well as the extract yield) were set, and then the probability-based design space was calculated by Monte Carlo simulation and verified. The results of the design space validation showed that the optimized purification method can ensure the stability of the Resina Draconis phenol extracts refining process, which would help to improve the quality uniformity between batches of phenol extracts and provide data support for production automation control. Copyright© by the Chinese Pharmaceutical Association.

  1. Evaporation of oil-water emulsion drops when heated at high temperature

    NASA Astrophysics Data System (ADS)

    Strizhak, P. A.; Piskunov, M. V.; Kuznetsov, G. V.; Voytkov, I. S.

    2017-10-01

    An experimental study on conditions and main characteristics for high-temperature (more than 700 K) evaporation of oil-water drops is presented. The high-temperature water purification from impurities can be the main practical application of research results. Thus, the heating of drops is implemented by the two typical schemes: on a massive substrate (the heating conditions are similar to those achieved in a heating chamber) and in a flow of the heated air. In the latter case, the heating conditions correspond to those attained while moving water drops with impurities in a counter high-temperature gaseous flow in the process of water purification. Evaporation time as function of heating temperature is presented. The influence of oil product concentration in an emulsion drop on evaporation characteristics is discussed. The conditions for intensive flash boiling of an emulsion drop and its explosive breakup with formation of the fine droplets cloud are pointed out. Heat fluxes required for intensive flash boiling and explosive breakup of a drop with further formation of the fine aerosol are determined in the boundary layer of a drop. The fundamental differences between flash boiling and explosive breakup of an emulsion drop when heated on a substrate and in a flow of the heated air are described. The main prospects for the development of the high-temperature water purification technology are detailed taking into account the fast emulsion drop breakup investigated in the paper.

  2. Characteristics of activated carbon produced from biosludge and its use in wastewater post-treatment.

    PubMed

    Pikkov, L; Kallas, J; Rüütmann, T; Rikmann, E

    2001-02-01

    Experimental research into the bench-scale production of activated carbon from waste-activated sludge from water purification, sawdust, peat, and their mixtures, by carbonisation and activation was undertaken. The research work was carried out to determine possible methods of production of cheap activated carbon from local raw materials and to use it in water purification technology. Along with the samples produced, several commercial activated carbons (namely RB-1, F 100, CA (adsorbent from military gas masks), BAY (product of the USSR)) were tested to compare adsorption properties in the adsorption of phenols, xylidines, amines, methylene blue and molasses. It has been found that the activated carbon produced from waste biosludge was of higher quality than that produced from either sawdust or peat, and performed similarly to RB-1 and F100 in adsorption tests. It was also determined that the activated carbon produced from biosludge could possibly be used in the post-treatment of wastewater. Residual sludge from the biological treatment of the wastewater from the purification of oil-shale in the chemical processing industry could cover up to 80% of the need for activated carbon. Some of this activated carbon could be used in the post-treatment of the same water, adsorbing polyalcaline phenols from the initial content of 4 mg l-1 to the demanded level of 1 mg l-1.

  3. Isolation and Purification of Water Soluble Proteins from Ginger Root (Zingiber officinale) by Two Dimensional Liquid Chromatography

    PubMed Central

    Sandovall, A.O.; Andrews, K.; Wahab, A.; Choudhary, M.I.; Ahmed, A.

    2014-01-01

    The RI-INBRE Centralized Core Facility was established in 2003 and participates annually in Undergraduate Summer Research Program. It provides students hands on research experience in key technologies in biomedical sciences. We present here the isolation and purification of water soluble proteins from ginger, a rhizome of the plant, Zingiber officinale. It is an important ingredient of species used in traditional South Asian cuisines. In Indian, Pakistani and Chinese folk medicine, ginger is used for gastro-intestinal disorders, nausea, vomiting, inflammatory diseases, muscle and joint pain. Limited studies have been reported on the bioactive proteins from ginger extract. The water soluble proteins were extracted from ginger root and successfully purified to homogeneity by using two-dimensional liquid chromatography (FPLC/RP-HPLC) approach. The ginger root was washed with distilled water; skin removed and then emulsified using an electric blender. Sample was stirred for four days at 4°C with and without protease inhibitor. Purification of a 42kDa protein was achieved by employing gel filtration, ion-exchange and reversed phase HPLC. The homogeneity of the protein was confirmed by SDS-PAGE gel electrophoresis and MALDI-TOF mass spectrometry. Future work will be conducted on the protein characterization using mass spectrometry and Edman protein sequencing. Supported by grant 5P20GM103430 from the National Institute of General Medical Sciences, NIH, USA.

  4. A flow-through chromatography process for influenza A and B virus purification.

    PubMed

    Weigel, Thomas; Solomaier, Thomas; Peuker, Alessa; Pathapati, Trinath; Wolff, Michael W; Reichl, Udo

    2014-10-01

    Vaccination is still the most efficient measure to protect against influenza virus infections. Besides the seasonal wave of influenza, pandemic outbreaks of bird or swine flu represent a high threat to human population. With the establishment of cell culture-based processes, there is a growing demand for robust, economic and efficient downstream processes for influenza virus purification. This study focused on the development of an economic flow-through chromatographic process avoiding virus strain sensitive capture steps. Therefore, a three-step process consisting of anion exchange chromatography (AEC), Benzonase(®) treatment, and size exclusion chromatography with a ligand-activated core (LCC) was established, and tested for purification of two influenza A virus strains and one influenza B virus strain. The process resulted in high virus yields (≥68%) with protein contamination levels fulfilling requirements of the European Pharmacopeia for production of influenza vaccines for human use. DNA was depleted by ≥98.7% for all strains. The measured DNA concentrations per dose were close to the required limits of 10ng DNA per dose set by the European Pharmacopeia. In addition, the added Benzonase(®) could be successfully removed from the product fraction. Overall, the presented downstream process could potentially represent a simple, robust and economic platform technology for production of cell culture-derived influenza vaccines. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. Rational design of peptide affinity ligands for the purification of therapeutic enzymes.

    PubMed

    Trasatti, John P; Woo, James; Ladiwala, Asif; Cramer, Steven; Karande, Pankaj

    2018-04-25

    Non-mAb biologics represent a growing class of therapeutics under clinical development. Although affinity chromatography is a potentially attractive approach for purification, the development of platform technologies, such as Protein A for mAbs, has been challenging due to the inherent chemical and structural diversity of these molecules. Here, we present our studies on the rapid development of peptide affinity ligands for the purification of biologics using a prototypical enzyme therapeutic in clinical use. Employing a suite of de novo rational and combinatorial design strategies we designed and screened a library of peptides on microarray platforms for their ability to bind to the target with high affinity and selectivity in cell culture fluid. Lead peptides were evaluated on resin in batch conditions and compared with a commercially available resin to evaluate their efficacy. Two lead candidates identified from microarray studies provided high binding capacity to the target while demonstrating high selectivity against culture contaminants and product variants compared to a commercial resin system. These findings provide a proof-of-concept for developing affinity peptide-based bioseparations processes for a target biologic. Peptide affinity ligand design and screening approaches presented in this work can also be easily translated to other biologics of interest. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 2018. © 2018 American Institute of Chemical Engineers.

  6. Continuous multistep synthesis of perillic acid from limonene by catalytic biofilms under segmented flow.

    PubMed

    Willrodt, Christian; Halan, Babu; Karthaus, Lisa; Rehdorf, Jessica; Julsing, Mattijs K; Buehler, Katja; Schmid, Andreas

    2017-02-01

    The efficiency of biocatalytic reactions involving industrially interesting reactants is often constrained by toxification of the applied biocatalyst. Here, we evaluated the combination of biologically and technologically inspired strategies to overcome toxicity-related issues during the multistep oxyfunctionalization of (R)-(+)-limonene to (R)-(+)-perillic acid. Pseudomonas putida GS1 catalyzing selective limonene oxidation via the p-cymene degradation pathway and recombinant Pseudomonas taiwanensis VLB120 were evaluated for continuous perillic acid production. A tubular segmented-flow biofilm reactor was used in order to relieve oxygen limitations and to enable membrane mediated substrate supply as well as efficient in situ product removal. Both P. putida GS1 and P. taiwanensis VLB120 developed a catalytic biofilm in this system. The productivity of wild-type P. putida GS1 encoding the enzymes for limonene bioconversion was highly dependent on the carbon source and reached 34 g L tube -1  day -1 when glycerol was supplied. More than 10-fold lower productivities were reached irrespective of the applied carbon source when the recombinant P. taiwanensis VLB120 harboring p-cymene monooxygenase and p-cumic alcohol dehydrogenase was used as biocatalyst. The technical applicability for preparative perillic acid synthesis in the applied system was verified by purification of perillic acid from the outlet stream using an anion exchanger resin. This concept enabled the multistep production of perillic acid and which might be transferred to other reactions involving volatile reactants and toxic end-products. Biotechnol. Bioeng. 2017;114: 281-290. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  7. Mining Critical Metals and Elements from Seawater: Opportunities and Challenges.

    PubMed

    Diallo, Mamadou S; Kotte, Madhusudhana Rao; Cho, Manki

    2015-08-18

    The availability and sustainable supply of technology metals and valuable elements is critical to the global economy. There is a growing realization that the development and deployment of the clean energy technologies and sustainable products and manufacturing industries of the 21st century will require large amounts of critical metals and valuable elements including rare-earth elements (REEs), platinum group metals (PGMs), lithium, copper, cobalt, silver, and gold. Advances in industrial ecology, water purification, and resource recovery have established that seawater is an important and largely untapped source of technology metals and valuable elements. This feature article discusses the opportunities and challenges of mining critical metals and elements from seawater. We highlight recent advances and provide an outlook of the future of metal mining and resource recovery from seawater.

  8. Research progress on quantum informatics and quantum computation

    NASA Astrophysics Data System (ADS)

    Zhao, Yusheng

    2018-03-01

    Quantum informatics is an emerging interdisciplinary subject developed by the combination of quantum mechanics, information science, and computer science in the 1980s. The birth and development of quantum information science has far-reaching significance in science and technology. At present, the application of quantum information technology has become the direction of people’s efforts. The preparation, storage, purification and regulation, transmission, quantum coding and decoding of quantum state have become the hotspot of scientists and technicians, which have a profound impact on the national economy and the people’s livelihood, technology and defense technology. This paper first summarizes the background of quantum information science and quantum computer and the current situation of domestic and foreign research, and then introduces the basic knowledge and basic concepts of quantum computing. Finally, several quantum algorithms are introduced in detail, including Quantum Fourier transform, Deutsch-Jozsa algorithm, Shor’s quantum algorithm, quantum phase estimation.

  9. Second International Conference on Accelerating Biopharmaceutical Development: March 9-12, 2009, Coronado, CA USA.

    PubMed

    Reichert, Janice M; Jacob, Nitya; Amanullah, Ashraf

    2009-01-01

    The Second International Conference on Accelerating Biopharmaceutical Development was held in Coronado, California. The meeting was organized by the Society for Biological Engineering (SBE) and the American Institute of Chemical Engineers (AIChE); SBE is a technological community of the AIChE. Bob Adamson (Wyeth) and Chuck Goochee (Centocor) were co-chairs of the event, which had the theme "Delivering cost-effective, robust processes and methods quickly and efficiently." The first day focused on emerging disruptive technologies and cutting-edge analytical techniques. Day two featured presentations on accelerated cell culture process development, critical quality attributes, specifications and comparability, and high throughput protein formulation development. The final day was dedicated to discussion of technology options and new analysis methods provided by emerging disruptive technologies; functional interaction, integration and synergy in platform development; and rapid and economic purification process development.

  10. Second International Conference on Accelerating Biopharmaceutical Development: March 9-12, 2009, Coronado, CA, USA.

    PubMed

    Reichert, Janice M; Jacob, Nitya M; Amanullah, Ashraf

    2009-01-01

    The Second International Conference on Accelerating Biopharmaceutical Development was held in Coronado, California. The meeting was organized by the Society for Biological Engineering (SBE) and the American Institute of Chemical Engineers (AIChE); SBE is a technological community of the AIChE. Bob Adamson (Wyeth) and Chuck Goochee (Centocor) were co-chairs of the event, which had the theme "Delivering cost-effective, robust processes and methods quickly and efficiently." The first day focused on emerging disruptive technologies and cutting-edge analytical techniques. Day two featured presentations on accelerated cell culture process development, critical quality attributes, specifications and comparability, and high throughput protein formulation development. The final day was dedicated to discussion of technology options and new analysis methods provided by emerging disruptive technologies; functional interaction, integration and synergy in platform development; and rapid and economic purification process development.

  11. Fully automated SPE-based synthesis and purification of 2-[18F]fluoroethyl-choline for human use.

    PubMed

    Schmaljohann, Jörn; Schirrmacher, Esther; Wängler, Björn; Wängler, Carmen; Schirrmacher, Ralf; Guhlke, Stefan

    2011-02-01

    2-[(18)F]Fluoroethyl-choline ([(18)F]FECH) is a promising tracer for the detection of prostate cancer as well as brain tumors with positron emission tomography (PET). [(18)F]FECH is actively transported into mammalian cells, becomes phosphorylated by choline kinase and gets incorporated into the cell membrane after being metabolized to phosphatidylcholine. So far, its synthesis is a two-step procedure involving at least one HPLC purification step. To allow a wider dissemination of this tracer, finding a purification method avoiding HPLC is highly desirable and would result in easier accessibility and more reliable production of [(18)F]FECH. [(18)F]FECH was synthesized by reaction of 2-bromo-1-[(18)F]fluoroethane ([(18)F]BFE) with dimethylaminoethanol (DMAE) in DMSO. We applied a novel and very reliable work-up procedure for the synthesis of [(18)F]BFE. Based on a combination of three different solid-phase cartridges, the purification of [(18)F]BFE from its precursor 2-bromoethyl-4-nitrobenzenesulfonate (BENos) could be achieved without using HPLC. Following the subsequent reaction of the purified [(18)F]BFE with DMAE, the final product [(18)F]FECH was obtained as a sterile solution by passing the crude reaction mixture through a combination of two CM plus cartridges and a sterile filter. The fully automated synthesis was performed using as well a Raytest SynChrom module (Raytest, Germany) or a Scintomics HotboxIII module (Scintomics, Germany). The radiotracer [(18)F]FECH can be synthesized in reliable radiochemical yields (RCY) of 37±5% (Synchrom module) and 33±5% (Hotbox III unit) in less than 1 h using these two fully automated commercially available synthesis units without HPLC involvement for purification. Detailed quality control of the final injectable [(18)F]FECH solution proved the high radiochemical purity and the absence of Kryptofix2.2.2, DMAE and DMSO used in the course of synthesis. Sterility and bacterial endotoxin testing following standard procedures verified that the described production method for [(18)F]FECH is suitable for human applications. The routine production of [(18)F]FECH with sufficient RCYs was established by reliable and fast solid-phase extraction purifications of both the secondary labeling precursor [(18)F]BFE and the final product [(18)F]FECH, avoiding complex and sensitive HPLC equipment. The purity of the product was >95%, rendering the tracer suitable for human application. The newly developed purification procedure for [(18)F]BFE significantly reduces the complexity of the automated synthesis unit, hence reducing the cost for routine production in a clinical setup and allowing easy transfer to different synthesis modules. Copyright © 2011 Elsevier Inc. All rights reserved.

  12. The bubble method of water purification

    NASA Astrophysics Data System (ADS)

    Smirnov, B. M.; Babaeva, N. Yu.; Naidis, G. V.; Panov, V. A.; Saveliev, A. S.; Son, E. E.; Tereshonok, D. V.

    2018-02-01

    The processes of water purification from admixture molecules are analyzed. The purification rate is limited due to a low diffusion coefficient of the admixture molecules in water. At non-small concentrations of the admixture molecules, the water purication can proceed through association of molecules in condensed nanoparticles which fall on the bottom of the water volume. The rate of association may be increased in an external electric field, but in reality this cannot change significantly the rate of the purification process. The bubble method of water purification is considered, where air bubbles formed at the bottom of the water volume, transfer admixture molecules to the interface. This method allows one to clean small water volumes fast. This mechanism of water purification is realized experimentally and exhibits the promises of the bubble purification method.

  13. Hofmeister series salts enhance purification of plasmid DNA by non-ionic detergents

    PubMed Central

    Lezin, George; Kuehn, Michael R.; Brunelli, Luca

    2011-01-01

    Ion-exchange chromatography is the standard technique used for plasmid DNA purification, an essential molecular biology procedure. Non-ionic detergents (NIDs) have been used for plasmid DNA purification, but it is unclear whether Hofmeister series salts (HSS) change the solubility and phase separation properties of specific NIDs, enhancing plasmid DNA purification. After scaling-up NID-mediated plasmid DNA isolation, we established that NIDs in HSS solutions minimize plasmid DNA contamination with protein. In addition, large-scale NID/HSS solutions eliminated LPS contamination of plasmid DNA more effectively than Qiagen ion-exchange columns. Large-scale NID isolation/NID purification generated increased yields of high quality DNA compared to alkali isolation/column purification. This work characterizes how HSS enhance NID-mediated plasmid DNA purification, and demonstrates that NID phase transition is not necessary for LPS removal from plasmid DNA. Specific NIDs such as IGEPAL CA-520 can be utilized for rapid, inexpensive and efficient laboratory-based large-scale plasmid DNA purification, outperforming Qiagen-based column procedures. PMID:21351074

  14. Isolation and purification of all-trans diadinoxanthin and all-trans diatoxanthin from diatom Phaeodactylum tricornutum.

    PubMed

    Kuczynska, Paulina; Jemiola-Rzeminska, Malgorzata

    2017-01-01

    Two diatom-specific carotenoids are engaged in the diadinoxanthin cycle, an important mechanism which protects these organisms against photoinhibition caused by absorption of excessive light energy. A high-performance and economical procedure of isolation and purification of diadinoxanthin and diatoxanthin from the marine diatom Phaeodactylum tricornutum using a four-step procedure has been developed. It is based on the use of commonly available materials and does not require advanced technology. Extraction of pigments, saponification, separation by partition and then open column chromatography, which comprise the complete experimental procedure, can be performed within 2 days. This method allows HPLC grade diadinoxanthin and diatoxanthin of a purity of 99 % or more to be obtained, and the efficiency was estimated to be 63 % for diadinoxanthin and 73 % for diatoxanthin. Carefully selected diatom culture conditions as well as analytical ones ensure highly reproducible performance. A protocol can be used to isolate and purify the diadinoxanthin cycle pigments both on analytical and preparative scale.

  15. Water management, purification, and conservation in arid climates. Volume 2: Water purification

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Goosen, M.F.A.; Shayya, W.H.

    1999-10-01

    Arid regions are already feeling the severe restraining effects of potable water shortages. In coming years, humid and sub-humid regions of the world will also have to face many of these same problems. In the future, serious conflicts may arise not because of a lack of oil, but due to water shortages. Are there solutions to these problems? Aside from increasing public awareness about the importance of water, society needs to take a three pronged approach: water needs to be effectively managed, it needs to be economically purified, and it needs to be conserved. Only by doing these three thingsmore » in unison can they hope to alleviate the water problems faced by arid regions of the world. This book presents information valuable to seeking, finding and using current technologies to help solve these problems now. Volume 2 presents various methods of purifying water, and includes membrane processes and alternative techniques such as solar desalination.« less

  16. Breakthrough in Xenon Capture and Purification Using Adsorbent-Supported Silver Nanoparticles.

    PubMed

    Deliere, Ludovic; Coasne, Benoit; Topin, Sylvain; Gréau, Claire; Moulin, Christophe; Farrusseng, David

    2016-07-04

    Rare gas capture and purification is a major challenge for energy, environment, and health applications. Of utmost importance for the nuclear industry, novel separation processes for Xe are urgently needed for spent nuclear fuel reprocessing and nuclear activity monitoring. The recovered, non-radioactive Xe is also of high economic value for lighting, surgical anesthetic, etc. Here, using adsorption and breakthrough experiments and statistical mechanics molecular simulation, we show the outstanding performance of zeolite-supported silver nanoparticles to capture/separate Xe at low concentrations (0.087-100 ppm). We also establish the efficiency of temperature swing adsorption based on such adsorbents for Xe separation from Kr/Xe mixtures and air streams corresponding to off-gases generated by nuclear reprocessing. This study paves the way for the development of novel, cost-efficient technologies relying on the large selectivity/capacity of adsorbent-supported silver nanoparticles which surpass all materials ever tested. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Versatile and on-demand biologics co-production in yeast.

    PubMed

    Cao, Jicong; Perez-Pinera, Pablo; Lowenhaupt, Ky; Wu, Ming-Ru; Purcell, Oliver; de la Fuente-Nunez, Cesar; Lu, Timothy K

    2018-01-08

    Current limitations to on-demand drug manufacturing can be addressed by technologies that streamline manufacturing processes. Combining the production of two or more drugs into a single batch could not only be useful for research, clinical studies, and urgent therapies but also effective when combination therapies are needed or where resources are scarce. Here we propose strategies to concurrently produce multiple biologics from yeast in single batches by multiplexing strain development, cell culture, separation, and purification. We demonstrate proof-of-concept for three biologics co-production strategies: (i) inducible expression of multiple biologics and control over the ratio between biologic drugs produced together; (ii) consolidated bioprocessing; and (iii) co-expression and co-purification of a mixture of two monoclonal antibodies. We then use these basic strategies to produce drug mixtures as well as to separate drugs. These strategies offer a diverse array of options for on-demand, flexible, low-cost, and decentralized biomanufacturing applications without the need for specialized equipment.

  18. 21 CFR 884.6170 - Assisted reproduction water and water purification systems.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Assisted reproduction water and water purification... Devices § 884.6170 Assisted reproduction water and water purification systems. (a) Identification. Assisted reproduction water purification systems are devices specifically intended to generate high quality...

  19. 21 CFR 884.6170 - Assisted reproduction water and water purification systems.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Assisted reproduction water and water purification... Devices § 884.6170 Assisted reproduction water and water purification systems. (a) Identification. Assisted reproduction water purification systems are devices specifically intended to generate high quality...

  20. 21 CFR 876.5665 - Water purification system for hemodialysis.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Water purification system for hemodialysis. 876... SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Therapeutic Devices § 876.5665 Water purification system for hemodialysis. (a) Identification. A water purification system for hemodialysis is a...

  1. 21 CFR 876.5665 - Water purification system for hemodialysis.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Water purification system for hemodialysis. 876... SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Therapeutic Devices § 876.5665 Water purification system for hemodialysis. (a) Identification. A water purification system for hemodialysis is a...

  2. 21 CFR 876.5665 - Water purification system for hemodialysis.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Water purification system for hemodialysis. 876... SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Therapeutic Devices § 876.5665 Water purification system for hemodialysis. (a) Identification. A water purification system for hemodialysis is a...

  3. 21 CFR 876.5665 - Water purification system for hemodialysis.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Water purification system for hemodialysis. 876... SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Therapeutic Devices § 876.5665 Water purification system for hemodialysis. (a) Identification. A water purification system for hemodialysis is a...

  4. 21 CFR 884.6170 - Assisted reproduction water and water purification systems.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Assisted reproduction water and water purification... Devices § 884.6170 Assisted reproduction water and water purification systems. (a) Identification. Assisted reproduction water purification systems are devices specifically intended to generate high quality...

  5. 21 CFR 884.6170 - Assisted reproduction water and water purification systems.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Assisted reproduction water and water purification... Devices § 884.6170 Assisted reproduction water and water purification systems. (a) Identification. Assisted reproduction water purification systems are devices specifically intended to generate high quality...

  6. 21 CFR 884.6170 - Assisted reproduction water and water purification systems.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Assisted reproduction water and water purification... Devices § 884.6170 Assisted reproduction water and water purification systems. (a) Identification. Assisted reproduction water purification systems are devices specifically intended to generate high quality...

  7. Magnetically controllable 3D microtissues based on magnetic microcryogels.

    PubMed

    Liu, Wei; Li, Yaqian; Feng, Siyu; Ning, Jia; Wang, Jingyu; Gou, Maling; Chen, Huijun; Xu, Feng; Du, Yanan

    2014-08-07

    Microtissues on the scale of several hundred microns are a promising cell culture configuration resembling the functional tissue units in vivo. In contrast to conventional cell culture, handling of microtissues poses new challenges such as medium exchange, purification and maintenance of the microtissue integrity. Here, we developed magnetic microcryogels to assist microtissue formation with enhanced controllability and robustness. The magnetic microcryogels were fabricated on-chip by cryogelation and micro-molding which could endure extensive external forces such as fluidic shear stress during pipetting and syringe injection. The magnetically controllable microtissues were applied to constitute a novel separable 3D co-culture system realizing functional enhancement of the hepatic microtissues co-cultured with the stromal microtissues and easy purification of the hepatic microtissues for downstream drug testing. The magnetically controllable microtissues with pre-defined shapes were also applied as building blocks to accelerate the tissue assembly process under magnetic force for bottom-up tissue engineering. Finally, the magnetic microcryogels could be injected in vivo as cell delivery vehicles and tracked by MRI. The injectable magnetic microtissues maintained viability at the injection site indicating good retention and potential applications for cell therapy. The magnetic microcryogels are expected to significantly promote the microtissues as a promising cellular configuration for cell-based applications such as in drug testing, tissue engineering and regenerative therapy.

  8. Treatment of PCR products with exonuclease I and heat-labile alkaline phosphatase improves the visibility of combined bisulfite restriction analysis

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Watanabe, Kousuke; Emoto, Noriko; Sunohara, Mitsuhiro

    2010-08-27

    Research highlights: {yields} Incubating PCR products at a high temperature causes smears in gel electrophoresis. {yields} Smears interfere with the interpretation of methylation analysis using COBRA. {yields} Treatment with exonuclease I and heat-labile alkaline phosphatase eliminates smears. {yields} The elimination of smears improves the visibility of COBRA. -- Abstract: DNA methylation plays a vital role in the regulation of gene expression. Abnormal promoter hypermethylation is an important mechanism of inactivating tumor suppressor genes in human cancers. Combined bisulfite restriction analysis (COBRA) is a widely used method for identifying the DNA methylation of specific CpG sites. Here, we report that exonucleasemore » I and heat-labile alkaline phosphatase can be used for PCR purification for COBRA, improving the visibility of gel electrophoresis after restriction digestion. This improvement is observed when restriction digestion is performed at a high temperature, such as 60 {sup o}C or 65 {sup o}C, with BstUI and TaqI, respectively. This simple method can be applied instead of DNA purification using spin columns or phenol/chloroform extraction. It can also be applied to other situations when PCR products are digested by thermophile-derived restriction enzymes, such as PCR restriction fragment length polymorphism (RFLP) analysis.« less

  9. Evaluation of strategies to control Fab light chain dimer during mammalian expression and purification: A universal one-step process for purification of correctly assembled Fab.

    PubMed

    Spooner, Jennifer; Keen, Jenny; Nayyar, Kalpana; Birkett, Neil; Bond, Nicholas; Bannister, David; Tigue, Natalie; Higazi, Daniel; Kemp, Benjamin; Vaughan, Tristan; Kippen, Alistair; Buchanan, Andrew

    2015-07-01

    Fabs are an important class of antibody fragment as both research reagents and therapeutic agents. There are a plethora of methods described for their recombinant expression and purification. However, these do not address the issue of excessive light chain production that forms light chain dimers nor do they describe a universal purification strategy. Light chain dimer impurities and the absence of a universal Fab purification strategy present persistent challenges for biotechnology applications using Fabs, particularly around the need for bespoke purification strategies. This study describes methods to address light chain dimer formation during Fab expression and identifies a novel CH 1 affinity resin as a simple and efficient one-step purification for correctly assembled Fab. © 2015 Wiley Periodicals, Inc.

  10. 24 CFR 203.52 - Acceptance of individual residential water purification equipment.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... residential water purification equipment. 203.52 Section 203.52 Housing and Urban Development Regulations... water purification equipment. If a property otherwise eligible for insurance under this part does not have access to a continuing supply of safe and potable water without the use of a water purification...

  11. 24 CFR 203.52 - Acceptance of individual residential water purification equipment.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... residential water purification equipment. 203.52 Section 203.52 Housing and Urban Development Regulations... water purification equipment. If a property otherwise eligible for insurance under this part does not have access to a continuing supply of safe and potable water without the use of a water purification...

  12. 24 CFR 203.52 - Acceptance of individual residential water purification equipment.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... residential water purification equipment. 203.52 Section 203.52 Housing and Urban Development Regulations... water purification equipment. If a property otherwise eligible for insurance under this part does not have access to a continuing supply of safe and potable water without the use of a water purification...

  13. 24 CFR 203.52 - Acceptance of individual residential water purification equipment.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... residential water purification equipment. 203.52 Section 203.52 Housing and Urban Development Regulations... water purification equipment. If a property otherwise eligible for insurance under this part does not have access to a continuing supply of safe and potable water without the use of a water purification...

  14. Chemical energy storage: Part of a systemic solution

    NASA Astrophysics Data System (ADS)

    Schlögl, Robert

    2017-07-01

    This paper is a primer into concepts and opportunities of chemical energy storage. Starting from the quest for decarbonisation we reveal the possibilities of chemical energy storage. We briefly discuss the critical role of catalysis as enabling technology. We concentrate on options of large-scale production of chemicals from CO2 and green hydrogen. We discuss one potential application of fueling future combustion engines that could run with minimal regulated emissions without exhaust purifications and legal tricks.

  15. Water Purification Characteristic of the Actual Constructed Wetland with Carex dispalata in a Cold Area

    NASA Astrophysics Data System (ADS)

    Tsuji, Morio; Yamada, Kazuhiro; Hiratsuka, Akira; Tsukada, Hiroko

    Carex dispalata, a native plant species applied in cold districts for water purification in constructed wetlands, has useful characteristics for landscape creation and maintenance. In this study, seasonal differences in purification ability were verified, along with comparison of frozen and non-frozen periods' performance. A wetland area was constructed using a “hydroponics method” and a “coir fiber based method”. Results show that the removal rates of BOD, SS, and Chl-a were high. On this constructed wetland reduces organic pollution, mainly phytoplankton, but the removal of nitrogen and phosphorus was insufficient. The respective mean values of influent and treated water during three years were 26.6 mg/L and 12.2 mg/L for BOD, and 27.9 mg/L and 7.5 mg/L for SS. The mean value of the BOD removal rate for the non-frozen period was 2.99 g/m2/d that for the frozen period was 1.86 g/m2/d. The removal rate followed the rise of the BOD load rate. The removal rate limits were about 4 g/m2/d during the frozen period and 15 g/m2/d during the non-frozen period. For operations, energy was unnecessary. The required working hours were about 20 h annually for all maintenance and management during operations.

  16. Host-Associated Metagenomics: A Guide to Generating Infectious RNA Viromes

    PubMed Central

    Robert, Catherine; Pascalis, Hervé; Michelle, Caroline; Jardot, Priscilla; Charrel, Rémi; Raoult, Didier; Desnues, Christelle

    2015-01-01

    Background Metagenomic analyses have been widely used in the last decade to describe viral communities in various environments or to identify the etiology of human, animal, and plant pathologies. Here, we present a simple and standardized protocol that allows for the purification and sequencing of RNA viromes from complex biological samples with an important reduction of host DNA and RNA contaminants, while preserving the infectivity of viral particles. Principal Findings We evaluated different viral purification steps, random reverse transcriptions and sequence-independent amplifications of a pool of representative RNA viruses. Viruses remained infectious after the purification process. We then validated the protocol by sequencing the RNA virome of human body lice engorged in vitro with artificially contaminated human blood. The full genomes of the most abundant viruses absorbed by the lice during the blood meal were successfully sequenced. Interestingly, random amplifications differed in the genome coverage of segmented RNA viruses. Moreover, the majority of reads were taxonomically identified, and only 7–15% of all reads were classified as “unknown”, depending on the random amplification method. Conclusion The protocol reported here could easily be applied to generate RNA viral metagenomes from complex biological samples of different origins. Our protocol allows further virological characterizations of the described viral communities because it preserves the infectivity of viral particles and allows for the isolation of viruses. PMID:26431175

  17. One-step purification of a functional, constitutively activated form of visual arrestin.

    PubMed

    Huang, Li; Mao, Xiang; Abdulaev, Najmoutin G; Ngo, Tony; Liu, Wei; Ridge, Kevin D

    2012-03-01

    Desensitization of agonist-activated G protein-coupled receptors (GPCRs) requires phosphorylation followed by the binding of arrestin, a ~48 kDa soluble protein. While crystal structures for the inactive, 'basal' state of various arrestins are available, the conformation of 'activated' arrestin adopted upon interaction with activated GPCRs remains unknown. As a first step towards applying high-resolution structural methods to study arrestin conformation and dynamics, we have utilized the subtilisin prodomain/Profinity eXact™ fusion-tag system for the high-level bacterial expression and one-step purification of wild-type visual arrestin (arrestin 1) as well as a mutant form (R175E) of the protein that binds to non-phosphorylated, light-activated rhodopsin (Rho∗). The results show that both prodomain/Profinity eXact™ fusion-tagged wild-type and R175E arrestins can be expressed to levels approaching 2-3 mg/l in Luria-Bertani media, and that the processed, tag-free mature forms can be purified to near homogeneity using a Bio-Scale™ Mini Profinity eXact™ cartridge on the Profinia™ purification system. Functional analysis of R175E arrestin generated using this approach shows that it binds to non-phosphorylated rhodopsin in a light-dependent manner. These findings should facilitate the structure determination of this 'constitutively activated' state of arrestin 1 as well as the monitoring of conformational changes upon interaction with Rho∗. Copyright © 2011 Elsevier Inc. All rights reserved.

  18. Hydrophilic interaction liquid chromatography for the separation, purification, and quantification of raffinose family oligosaccharides from Lycopus lucidus Turcz.

    PubMed

    Liang, Tu; Fu, Qing; Li, Fangbing; Zhou, Wei; Xin, Huaxia; Wang, Hui; Jin, Yu; Liang, Xinmiao

    2015-08-01

    A systematic strategy based on hydrophilic interaction liquid chromatography was developed for the separation, purification and quantification of raffinose family oligosaccharides from Lycopus lucidus Turcz. Methods with enough hydrophilicity and selectivity were utilized to resolve the problems encountered in the separation of oligosaccharides such as low retention, low resolution and poor solubility. The raffinose family oligosaccharides in L. lucidus Turcz. were isolated using solid-phase extraction followed by hydrophilic interaction liquid chromatography at semi-preparative scale to obtain standards of stachyose, verbascose and ajugose. Utilizing the obtained oligosaccharides as standards, a quantitative determination method was developed, validated and applied for the content determination of raffinose family oligosaccharides both in the aerial and root parts of L. lucidus Turcz. There were no oligosaccharides in the aerial parts, while in the root parts, the total content was 686.5 mg/g with the average distribution: raffinose 66.5 mg/g, stachyose 289.0 mg/g, verbascose 212.4 mg/g, and ajugose 118.6 mg/g. The result provided the potential of roots of L. lucidus Turcz. as new raffinose family oligosaccharides sources for functional food. Moreover, since the present systematic strategy is efficient, sensitive and robust, separation, purification and quantification of oligosaccharides by hydrophilic interaction liquid chromatography seems to be possible. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Entanglement of purification: from spin chains to holography

    NASA Astrophysics Data System (ADS)

    Nguyen, Phuc; Devakul, Trithep; Halbasch, Matthew G.; Zaletel, Michael P.; Swingle, Brian

    2018-01-01

    Purification is a powerful technique in quantum physics whereby a mixed quantum state is extended to a pure state on a larger system. This process is not unique, and in systems composed of many degrees of freedom, one natural purification is the one with minimal entanglement. Here we study the entropy of the minimally entangled purification, called the entanglement of purification, in three model systems: an Ising spin chain, conformal field theories holographically dual to Einstein gravity, and random stabilizer tensor networks. We conjecture values for the entanglement of purification in all these models, and we support our conjectures with a variety of numerical and analytical results. We find that such minimally entangled purifications have a number of applications, from enhancing entanglement-based tensor network methods for describing mixed states to elucidating novel aspects of the emergence of geometry from entanglement in the AdS/CFT correspondence.

  20. An effective purification method using large bottles for human pancreatic islet isolation

    PubMed Central

    Shimoda, Masayuki; Itoh, Takeshi; Iwahashi, Shuichi; Takita, Morihito; Sugimoto, Koji; Kanak, Mazhar A.; Chujo, Daisuke; Naziruddin, Bashoo; Levy, Marlon F.; Grayburn, Paul A.; Matsumoto, Shinichi

    2012-01-01

    The purification process is one of the most difficult procedures in pancreatic islet isolation. It was demonstrated that the standard purification method using a COBE 2991 cell processor with Ficoll density gradient solution harmed islets mechanically by high shear force. We reported that purification using large bottles with a lower viscosity gradient solution could improve the efficacy of porcine islet purification. In this study, we examined whether the new bottle purification method could improve the purification of human islets. Nine human pancreata from brain-dead donors were used. After pancreas digestion, the digested tissue was divided into three groups. Each group was purified by continuous density gradient using ET-Kyoto and iodixanol gradient solution with either the standard COBE method (COBE group) or the top loading (top group) or bottom loading (bottom group) bottle purification methods. Islet yield, purity, recovery rate after purification, and in vitro and in vivo viability were compared. Islet yield per pancreas weight (IE/g) and the recovery rate in the top group were significantly higher than in the COBE and bottom groups. Furthermore, the average size of purified islets in the top group was significantly larger than in the COBE group, which indicated that the bottle method could reduce the shear force to the islets. In vivo viability was also significantly higher in the top group compared with the COBE group. In conclusion, the top-loading bottle method could improve the quality and quantity of human islets after purification. PMID:23221740

  1. Electron Beam Technology for Environmental Pollution Control.

    PubMed

    Chmielewski, Andrzej G; Han, Bumsoo

    2016-10-01

    Worldwide, there are over 1700 electron beam (EB) units in commercial use, providing an estimated added value to numerous products, amounting to 100 billion USD or more. High-current electron accelerators are used in diverse industries to enhance the physical and chemical properties of materials and to reduce undesirable contaminants such as pathogens, toxic byproducts, or emissions. Over the past few decades, EB technologies have been developed aimed at ensuring the safety of gaseous and liquid effluents discharged to the environment. It has been demonstrated that EB technologies for flue gas treatment (SO x and NO x removal), wastewater purification, and sludge hygienization can be effectively deployed to mitigate environmental degradation. Recently, extensive work has been carried out on the use of EB for environmental remediation, which also includes the removal of emerging contaminants such as VOCs, endocrine disrupting chemicals (EDCs), and potential EDCs.

  2. Immunoglobulin domain interface exchange as a platform technology for the generation of Fc heterodimers and bispecific antibodies.

    PubMed

    Skegro, Darko; Stutz, Cian; Ollier, Romain; Svensson, Emelie; Wassmann, Paul; Bourquin, Florence; Monney, Thierry; Gn, Sunitha; Blein, Stanislas

    2017-06-09

    Bispecific antibodies (bsAbs) are of significant importance to the development of novel antibody-based therapies, and heavy chain (Hc) heterodimers represent a major class of bispecific drug candidates. Current technologies for the generation of Hc heterodimers are suboptimal and often suffer from contamination by homodimers posing purification challenges. Here, we introduce a new technology based on biomimicry wherein the protein-protein interfaces of two different immunoglobulin (Ig) constant domain pairs are exchanged in part or fully to design new heterodimeric domains. The method can be applied across Igs to design Fc heterodimers and bsAbs. We investigated interfaces from human IgA CH3, IgD CH3, IgG1 CH3, IgM CH4, T-cell receptor (TCR) α/β, and TCR γ/δ constant domain pairs, and we found that they successfully drive human IgG1 CH3 or IgM CH4 heterodimerization to levels similar to or above those of reference methods. A comprehensive interface exchange between the TCR α/β constant domain pair and the IgG1 CH3 homodimer was evidenced by X-ray crystallography and used to engineer examples of bsAbs for cancer therapy. Parental antibody pairs were rapidly reformatted into scalable bsAbs that were free of homodimer traces by combining interface exchange, asymmetric Protein A binding, and the scFv × Fab format. In summary, we successfully built several new CH3- or CH4-based heterodimers that may prove useful for designing new bsAb-based therapeutics, and we anticipate that our approach could be broadly implemented across the Ig constant domain family. To our knowledge, CH4-based heterodimers have not been previously reported. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  3. Microfabricated Genomic Analysis System

    NASA Technical Reports Server (NTRS)

    Gonda, Steve; Elms, Rene

    2005-01-01

    Genetic sequencing and many genetic tests and assays require electrophoretic separation of DNA. In this technique, DNA fragments are separated by size as they migrate through a sieving gel under the influence of an applied electric field. In order to conduct these analyses on-orbit, it is essential to acquire the capability to efficiently perform electrophoresis in a microgravity environment. Conventional bench top electrophoresis equipment is large and cumbersome and does not lead itself to on-orbit utilization. Much of the previous research regarding on-orbit electrophoresis involved altering conventional electrophoresis equipment for bioprocessing, purification, and/or separation technology applications. A new and more efficient approach to on-orbit electrophoresis is the use of a microfabricated electrophoresis platform. These platforms are much smaller, less expensive to produce and operate, use less power, require smaller sample sizes (nanoliters), and achieve separation in a much shorter distance (a few centimeters instead of 10 s or 100 s of centimeters.) In contrast to previous applications, this platform would be utilized as an analytical tool for life science/medical research, environmental monitoring, and medical diagnoses. Identification of infectious agents as well as radiation related damage are significant to NASA s efforts to maintain, study, and monitor crew health during and in support of near-Earth and interplanetary missions. The capability to perform genetic assays on-orbit is imperative to conduct relevant and insightful biological and medical research, as well as continuing NASA s search for life elsewhere. This technology would provide an essential analytical tool for research conducted in a microgravity environment (Shuttle, ISS, long duration/interplanetary missions.) In addition, this technology could serve as a critical and invaluable component of a biosentinel system to monitor space environment genotoxic insults to include radiation.

  4. Efficient purification of paclitaxel from yews using high-performance displacement chromatography technique.

    PubMed

    Watchueng, Jean; Kamnaing, Pierre; Gao, Jin-Ming; Kiyota, Taira; Yeboah, Faustinus; Konishi, Yasuo

    2011-05-20

    Paclitaxel was purified using high-performance displacement chromatography (HPDC) technique, but not by the mechanism of HPDC. On small scale, paclitaxel was extracted with methanol from dry needles of Taxus canadensis and was enriched by extracting with chloroform after removing water-soluble hydrophilic components and hexane-soluble hydrophobic components. Then, 93-99% purity of paclitaxel was obtained using the HPDC technique. On large scale, taxanes were enriched by solvent partitioning between acetic acid/MeOH/H(2)O and hexane and extracted with CH(2)Cl(2). Taxanes except paclitaxel were further removed by extracting with methanol-water-trifluoroacetic acid (1.0:98.9:0.1, v/v/v). Applying HPDC technique to water-insoluble substances is problematic as this method requires a highly aqueous solvent system. In order to overcome this incompatibility, a system was set up where paclitaxel, although in low concentration, was extracted by methanol-water-trifluoroacetic acid (10.0:89.9:0.1, v/v/v). Recycling the extracting solvent to ensure minimal volume, the extracted paclitaxel was adsorbed on a C(18) trap column. A C(18) column of 4.6mm internal diameter was then connected to the trap column. The HPDC technique was thus carried out using an isocratic acetonitrile-water-trifluoroacetic acid (30.0:69.9:0.1, v/v/v) mobile phase consisting of a displacer cetylpyridinium trifluoroacetate (3mg/mL). Paclitaxel was co-eluted with the displacer and spontaneously crystallized. The crystal (114mg) showed 99.4% purity and only 10% of paclitaxel in the starting crude extract was lost during the enrichment/purification processes. This large scale purification method was successfully applied to purify paclitaxel from Chinese yew in small scale, suggesting general applicability of the method. This is the first report of purifying a water-insoluble natural product using HPDC technique. Crown Copyright © 2011. Published by Elsevier B.V. All rights reserved.

  5. Preparation of Protein Samples for NMR Structure, Function, and Small Molecule Screening Studies

    PubMed Central

    Acton, Thomas B.; Xiao, Rong; Anderson, Stephen; Aramini, James; Buchwald, William A.; Ciccosanti, Colleen; Conover, Ken; Everett, John; Hamilton, Keith; Huang, Yuanpeng Janet; Janjua, Haleema; Kornhaber, Gregory; Lau, Jessica; Lee, Dong Yup; Liu, Gaohua; Maglaqui, Melissa; Ma, Lichung; Mao, Lei; Patel, Dayaban; Rossi, Paolo; Sahdev, Seema; Shastry, Ritu; Swapna, G.V.T.; Tang, Yeufeng; Tong, Saichiu; Wang, Dongyan; Wang, Huang; Zhao, Li; Montelione, Gaetano T.

    2014-01-01

    In this chapter, we concentrate on the production of high quality protein samples for NMR studies. In particular, we provide an in-depth description of recent advances in the production of NMR samples and their synergistic use with recent advancements in NMR hardware. We describe the protein production platform of the Northeast Structural Genomics Consortium, and outline our high-throughput strategies for producing high quality protein samples for nuclear magnetic resonance (NMR) studies. Our strategy is based on the cloning, expression and purification of 6X-His-tagged proteins using T7-based Escherichia coli systems and isotope enrichment in minimal media. We describe 96-well ligation-independent cloning and analytical expression systems, parallel preparative scale fermentation, and high-throughput purification protocols. The 6X-His affinity tag allows for a similar two-step purification procedure implemented in a parallel high-throughput fashion that routinely results in purity levels sufficient for NMR studies (> 97% homogeneity). Using this platform, the protein open reading frames of over 17,500 different targeted proteins (or domains) have been cloned as over 28,000 constructs. Nearly 5,000 of these proteins have been purified to homogeneity in tens of milligram quantities (see Summary Statistics, http://nesg.org/statistics.html), resulting in more than 950 new protein structures, including more than 400 NMR structures, deposited in the Protein Data Bank. The Northeast Structural Genomics Consortium pipeline has been effective in producing protein samples of both prokaryotic and eukaryotic origin. Although this paper describes our entire pipeline for producing isotope-enriched protein samples, it focuses on the major updates introduced during the last 5 years (Phase 2 of the National Institute of General Medical Sciences Protein Structure Initiative). Our advanced automated and/or parallel cloning, expression, purification, and biophysical screening technologies are suitable for implementation in a large individual laboratory or by a small group of collaborating investigators for structural biology, functional proteomics, ligand screening and structural genomics research. PMID:21371586

  6. Microchip-based cell lysis and DNA extraction from sperm cells for application to forensic analysis.

    PubMed

    Bienvenue, Joan M; Duncalf, Natalie; Marchiarullo, Daniel; Ferrance, Jerome P; Landers, James P

    2006-03-01

    The current backlog of casework is among the most significant challenges facing crime laboratories at this time. While the development of next-generation microchip-based technology for expedited forensic casework analysis offers one solution to this problem, this will require the adaptation of manual, large-volume, benchtop chemistry to small volume microfluidic devices. Analysis of evidentiary materials from rape kits where semen or sperm cells are commonly found represents a unique set of challenges for on-chip cell lysis and DNA extraction that must be addressed for successful application. The work presented here details the development of a microdevice capable of DNA extraction directly from sperm cells for application to the analysis of sexual assault evidence. A variety of chemical lysing agents are assessed for inclusion in the extraction protocol and a method for DNA purification from sperm cells is described. Suitability of the extracted DNA for short tandem repeat (STR) analysis is assessed and genetic profiles shown. Finally, on-chip cell lysis methods are evaluated, with results from fluorescence visualization of cell rupture and DNA extraction from an integrated cell lysis and purification with subsequent STR amplification presented. A method for on-chip cell lysis and DNA purification is described, with considerations toward inclusion in an integrated microdevice capable of both differential cell sorting and DNA extraction. The results of this work demonstrate the feasibility of incorporating microchip-based cell lysis and DNA extraction into forensic casework analysis.

  7. Nanomagnet-based removal of lead and digoxin from living rats

    NASA Astrophysics Data System (ADS)

    Herrmann, Inge K.; Schlegel, Andrea; Graf, Rolf; Schumacher, Christoph M.; Senn, Nico; Hasler, Melanie; Gschwind, Sabrina; Hirt, Ann-Marie; Günther, Detlef; Clavien, Pierre-Alain; Stark, Wendelin J.; Beck-Schimmer, Beatrice

    2013-08-01

    In a number of clinical conditions such as intoxication, bacteraemia or autoimmune diseases the removal of the disease-causing factor from blood would be the most direct cure. However, physicochemical characteristics of the target compounds limit the applicability of classical filtration and diffusion-based processes. In this work, we present a first in vivo magnetic blood purification rodent animal model and demonstrate its ability to rapidly clear toxins from blood circulation using two model toxins with stable plasma levels (lead (Pb2+) and digoxin). Ultra-strong functionalized metal nanomagnets are employed to eliminate the toxin from whole blood in an extracorporeal circuit. In the present experimental demonstration over 40% of the toxin (i.e. lead or digoxin) was removed within the first 10 minutes and over 75% within 40 minutes. After capturing the target substance, a magnetic trap prevents the toxin-loaded nanoparticles from entering the blood circulation. Elemental analysis and magnetic hysteresis measurements confirm full particle recovery by simple magnetic separation (residual particle concentration below 1 μg mL-1 (detection limit)). We demonstrate that magnetic separation-based blood purification offers rapid blood cleaning from noxious agents, germs or other deleterious materials with relevance to a number of clinical conditions. Based on this new approach, current blood purification technologies can be extended to efficiently remove disease-causing factors, e.g. overdosed drugs, bacteria or cancer cells without being limited by filter cut-offs or column surface saturation.

  8. Mapping protein-RNA interactions by RCAP, RNA-cross-linking and peptide fingerprinting.

    PubMed

    Vaughan, Robert C; Kao, C Cheng

    2015-01-01

    RNA nanotechnology often feature protein RNA complexes. The interaction between proteins and large RNAs are difficult to study using traditional structure-based methods like NMR or X-ray crystallography. RCAP, an approach that uses reversible-cross-linking affinity purification method coupled with mass spectrometry, has been developed to map regions within proteins that contact RNA. This chapter details how RCAP is applied to map protein-RNA contacts within virions.

  9. Iodine Disinfection in the Use of Individual Water Purification Devices

    DTIC Science & Technology

    2006-03-01

    Effect of Resin Disinfectants-I3 and –I5 on Giardia muris and Giardia lamblia. Applied and Environmental Microbiology, 46(5), 965-969. 24. AWWA... Giardia or Cryptosporidium). Iodine-using IWPDs meeting these standards are considered effective against disease causing bacteria, viruses, and...CT = 65 mg-min/L) for a 2-log inactivation of E. histolytica cysts (references 9 and 10). Another study using Giardia cysts showed CT’s up to 3

  10. Capacity on wireless quantum cellular communication system

    NASA Astrophysics Data System (ADS)

    Zhou, Xiang-Zhen; Yu, Xu-Tao; Zhang, Zai-Chen

    2018-03-01

    Quantum technology is making excellent prospects in future communication networks. Entanglement generation and purification are two major components in quantum networks. Combining these two techniques with classical cellular mobile communication, we proposed a novel wireless quantum cellular(WQC) communication system which is possible to realize commercial mobile quantum communication. In this paper, the architecture and network topology of WQC communication system are discussed, the mathematical model of WQC system is extracted and the serving capacity, indicating the ability to serve customers, is defined and calculated under certain circumstances.

  11. Diastereoselective chain-elongation reactions using microreactors for applications in complex molecule assembly.

    PubMed

    Carter, Catherine F; Lange, Heiko; Sakai, Daiki; Baxendale, Ian R; Ley, Steven V

    2011-03-14

    Diastereoselective chain-elongation reactions are important transformations for the assembly of complex molecular structures, such as those present in polyketide natural products. Here we report new methods for performing crotylation reactions and homopropargylation reactions by using newly developed low-temperature flow-chemistry technology. In-line purification protocols are described, as well as the application of the crotylation protocol in an automated multi-step sequence. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Necessity of purification during bacterial DNA extraction with environmental soils

    PubMed Central

    Choi, Jung-Hyun

    2017-01-01

    Complexity and heterogeneity of soil samples have often implied the inclusion of purification steps in conventional DNA extraction for polymerase chain reaction (PCR) assays. Unfortunately the purification steps are also time and labor intensive. Therefore the necessity of DNA purification was re-visited and investigated for a variety of environmental soil samples that contained various amounts of PCR inhibitors. Bead beating and centrifugation was used as the baseline (without purification) method for DNA extraction. Its performance was compared with that of conventional DNA extraction kit (with purification). The necessity criteria for DNA purification were established with environmental soil samples. Using lysis conditions at 3000 rpm for 3 minutes with 0.1 mm glass beads, centrifugation time of 10 minutes and 1:10 dilution ratio, the baseline method outperformed conventional DNA extraction on cell seeded sand samples. Further investigation with PCR inhibitors (i.e., humic acids, clay, and magnesium [Mg]) showed that sand samples containing less than 10 μg/g humic acids and 70% clay may not require purifications. Interestingly, the inhibition pattern of Mg ion was different from other inhibitors due to the complexation interaction of Mg ion with DNA fragments. It was concluded that DNA extraction method without purification is suitable for soil samples that have less than 10 μg/g of humic acids, less than 70% clay content and less than 0.01% Mg ion content. PMID:28793754

  13. The ADESORB Process for Economical Production of Sorbents for Mercury Removal from Coal Fired Power Plants

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Robin Stewart

    The DOE's National Energy Technology Laboratory (NETL) currently manages the largest research program in the country for controlling coal-based mercury emissions. NETL has shown through various field test programs that the determination of cost-effective mercury control strategies is complex and highly coal- and plant-specific. However, one particular technology has the potential for widespread application: the injection of activated carbon upstream of either an electrostatic precipitator (ESP) or a fabric filter baghouse. This technology has potential application to the control of mercury emissions on all coal-fired power plants, even those with wet and dry scrubbers. This is a low capital costmore » technology in which the largest cost element is the cost of sorbents. Therefore, the obvious solutions for reducing the costs of mercury control must focus on either reducing the amount of sorbent needed or decreasing the cost of sorbent production. NETL has researched the economics and performance of novel sorbents and determined that there are alternatives to the commercial standard (NORIT DARCO{reg_sign} Hg) and that this is an area where significant technical improvements can still be made. In addition, a key barrier to the application of sorbent injection technology to the power industry is the availability of activated carbon production. Currently, about 450 million pounds ($250 million per year) of activated carbon is produced and used in the U.S. each year - primarily for purification of drinking water, food, and beverages. If activated carbon technology were to be applied to all 1,100 power plants, EPA and DOE estimate that it would require an additional $1-$2 billion per year, which would require increasing current capacity by a factor of two to eight. A new facility to produce activated carbon would cost approximately $250 million, would increase current U.S. production by nearly 25%, and could take four to five years to build. This means that there could be significant shortages in supply if response to new demand is not well-timed.« less

  14. Assessment of the microbial integrity, sensu G.S. Wilson, of piped and bottled drinking water in the condition as ingested.

    PubMed

    Mossel, David A A; Struijk, Corry B

    2004-05-01

    The second half of the 20th century witnessed substantial progress in the assurance and verification of microbiological integrity, i.e., safety and sensory quality, of drinking water. Enteropathogenic agents, such as particular viruses and protozoa, not previously identified as transmitted by industrially provided water supplies, were demonstrated to cause disease outbreaks, when ingested with piped water. The potential harm posed by carry-over of orally toxic metabolites of organisms, producing 'algal' (cyanophytic) blooms, was considered. In addition, earlier observations on the colonization of attenuated drinking water bodies by a variety of oligotrophic Gram-negative bacteria were confirmed and extended. This new evidence called for updating both water purification technologies and analytical methodology, serving to verify that goals had been attained. For the former purpose, the hazard analysis empowering control of critical practices (HACCP) strategy, introduced about 1960 in industrial food processing, was successfully adopted. Elimination, devitalization or barrier technologies for the more recently identified water-borne pathogens were elaborated, taking account of the hazard of production of chlorinated compounds with alleged adverse health effects. Biofilm formation throughout water distribution networks was brought under control by strict limitation of concentrations of compounds, assimilable by oligotrophic bacteria. Upon acknowledging that direct detection tests for pathogens were futile, because of their most sporadic and erratic distribution, Schardinger's marker organism concept was anew embraced, rigorously revised and substantially enlarged. Misleading designations, like searches for 'faecal coliforms' were replaced by boundary testing for Escherichia coli and appropriate Enterococcus spp. In addition, though still to be perfected, detection protocols for relevant bacteriophages or index viruses and, to a certain extent, also for spores of aerobic and anaerobic sporing rods were also elaborated. In all monitoring account was taken of sublethally injured target organisms, surviving purification technologies, though not deprived of their ecological significance. A need remains for a rigorously standardized operating procedure (SOP) for colony counts of psychrotrophic, oligotrophic Gram-negative rod-shaped bacteria ('heterotrophic plate count'), which constitute a useful criterion of indicator value. As in the contemporary HACCP approach to food safety, guidelines for assessing success or failure in control of integrity (Water Safety Objectives) were empirically elaborated. These rely on surveys on water samples, originating from drinking water supplies, previously verified as complying with longitudinally integrated HACCP-based purification technologies. Structured Academic dissemination of these innovations, through professional microbiologists to operator and executive levels, is recommended. Web based Distance Learning MSc Programmes, like the one, since the academic year 2003-2004, offered by the University of Hertfordshire, Hatfield, UK, may contribute to such endeavours. Though the complete Course is centered around Food Safety, the Modules in-Residence Practicals and Science and Technology of Drinking Water can be studied as an entity while being employed. Copyright 2004 Elsevier B.V.

  15. Proteomic Investigations into Hemodialysis Therapy

    PubMed Central

    Bonomini, Mario; Sirolli, Vittorio; Pieroni, Luisa; Felaco, Paolo; Amoroso, Luigi; Urbani, Andrea

    2015-01-01

    The retention of a number of solutes that may cause adverse biochemical/biological effects, called uremic toxins, characterizes uremic syndrome. Uremia therapy is based on renal replacement therapy, hemodialysis being the most commonly used modality. The membrane contained in the hemodialyzer represents the ultimate determinant of the success and quality of hemodialysis therapy. Membrane’s performance can be evaluated in terms of removal efficiency for unwanted solutes and excess fluid, and minimization of negative interactions between the membrane material and blood components that define the membrane’s bio(in)compatibility. Given the high concentration of plasma proteins and the complexity of structural functional relationships of this class of molecules, the performance of a membrane is highly influenced by its interaction with the plasma protein repertoire. Proteomic investigations have been increasingly applied to describe the protein uremic milieu, to compare the blood purification efficiency of different dialyzer membranes or different extracorporeal techniques, and to evaluate the adsorption of plasma proteins onto hemodialysis membranes. In this article, we aim to highlight investigations in the hemodialysis setting making use of recent developments in proteomic technologies. Examples are presented of why proteomics may be helpful to nephrology and may possibly affect future directions in renal research. PMID:26690416

  16. Supercritical fluid processing: opportunities for new resist materials and processes

    NASA Astrophysics Data System (ADS)

    Gallagher-Wetmore, Paula M.; Ober, Christopher K.; Gabor, Allen H.; Allen, Robert D.

    1996-05-01

    Over the past two decades supercritical fluids have been utilized as solvents for carrying out separations of materials as diverse as foods, polymers, pharmaceuticals, petrochemicals, natural products, and explosives. More recently they have been used for non-extractive applications such as recrystallization, deposition, impregnation, surface modification, and as a solvent alternative for precision parts cleaning. Today, supercritical fluid extraction is being practiced in the foods and beverage industries; there are commercial plants for decaffeinating coffee and tea, extracting beer flavoring agents from hops, and separating oils and oleoresins from spices. Interest in supercritical fluid processing of polymers has grown over the last ten years, and many new purification, fractionation, and even polymerization techniques have emerged. One of the most significant motivations for applying this technology to polymers has been increased performance demands. More recently, with increasing scrutiny of traditional solvents, supercritical fluids, and in particular carbon dioxide, are receiving widespread attention as 'environmentally conscious' solvents. This paper describes several examples of polymers applications, including a few involving photoresists, which demonstrate that as next- generation advanced polymer systems emerge, supercritical fluids are certain to offer advantages as cutting edge processing tools.

  17. Structure Analysis of Effective Chemical Compounds against Dengue Viruses Isolated from Isatis tinctoria.

    PubMed

    Gao, Bo; Zhang, Jianming; Xie, Lianhui

    2018-01-01

    The history of Chinese herb research can be traced back to thousands of years ago, and the abundant knowledge accumulated for these herbs makes them good candidates for developing new natural drugs. Isatis tinctoria is probably the most well-studied Chinese herb, which has been identified to be effective against dengue fever. However, the underlying biological mechanisms are still unclear. In this study, we adopt combined methods of bioactive trace technology and phytochemical extraction and separation, to guide the isolation and purification of the effective chemical constituents on the water-soluble components of aerial parts of Isatis tinctoria . In addition, we apply polarimetry and 1D or 2D nuclear magnetic resonance (NMR) spectroscopy to identify their structures, which lay a foundation for further study on the biological mechanisms underlying medicinal effects of Isatis tinctoria using in vitro and in vivo experiments. Specifically, we identify and infer the structures of 27 types of chemical compounds named GB-1, GB-2, …, GB-27, respectively, among which GB-7 is a novel compound. Further study of these compounds is critical to reveal the secrets behind the medicinal effects of Isatis tinctoria .

  18. Structure Analysis of Effective Chemical Compounds against Dengue Viruses Isolated from Isatis tinctoria

    PubMed Central

    Gao, Bo; Zhang, Jianming

    2018-01-01

    The history of Chinese herb research can be traced back to thousands of years ago, and the abundant knowledge accumulated for these herbs makes them good candidates for developing new natural drugs. Isatis tinctoria is probably the most well-studied Chinese herb, which has been identified to be effective against dengue fever. However, the underlying biological mechanisms are still unclear. In this study, we adopt combined methods of bioactive trace technology and phytochemical extraction and separation, to guide the isolation and purification of the effective chemical constituents on the water-soluble components of aerial parts of Isatis tinctoria. In addition, we apply polarimetry and 1D or 2D nuclear magnetic resonance (NMR) spectroscopy to identify their structures, which lay a foundation for further study on the biological mechanisms underlying medicinal effects of Isatis tinctoria using in vitro and in vivo experiments. Specifically, we identify and infer the structures of 27 types of chemical compounds named GB-1, GB-2, …, GB-27, respectively, among which GB-7 is a novel compound. Further study of these compounds is critical to reveal the secrets behind the medicinal effects of Isatis tinctoria. PMID:29808104

  19. Advances in identification and validation of protein targets of natural products without chemical modification.

    PubMed

    Chang, J; Kim, Y; Kwon, H J

    2016-05-04

    Covering: up to February 2016Identification of the target proteins of natural products is pivotal to understanding the mechanisms of action to develop natural products for use as molecular probes and potential therapeutic drugs. Affinity chromatography of immobilized natural products has been conventionally used to identify target proteins, and has yielded good results. However, this method has limitations, in that labeling or tagging for immobilization and affinity purification often result in reduced or altered activity of the natural product. New strategies have recently been developed and applied to identify the target proteins of natural products and synthetic small molecules without chemical modification of the natural product. These direct and indirect methods for target identification of label-free natural products include drug affinity responsive target stability (DARTS), stability of proteins from rates of oxidation (SPROX), cellular thermal shift assay (CETSA), thermal proteome profiling (TPP), and bioinformatics-based analysis of connectivity. This review focuses on and reports case studies of the latest advances in target protein identification methods for label-free natural products. The integration of newly developed technologies will provide new insights and highlight the value of natural products for use as biological probes and new drug candidates.

  20. Enzymatic pulp upgrade for producing high-value cellulose out of a Kraft paper pulp.

    PubMed

    Hutterer, Christian; Kliba, Gerhard; Punz, Manuel; Fackler, Karin; Potthast, Antje

    2017-07-01

    The high-yield separation of polymeric parts from wood-derived lignocellulosic material is indispensable in biorefinery concepts. For the separation of cellulose and xylan from hardwood paper pulps to obtain pulps of high cellulose contents, simple alkaline extractions were found to be the most suitable technology, although having certain limitations. These are embodied by residual alkali resistant xylan incorporated in the pulp matrix. Further purification in order to produce pure cellulose with a low uniformity could be achieved selectively degrading residual xylan and depolymerizing the cellulose macromolecules by xylanase and cellulase. The latter help to adjust cellulose chain lengths for certain dissolving pulp grades while reducing the demand for ozone in subsequent TCF bleaching. Experiments applying different commercially available enzyme preparations revealed the dependency of xylanase performance on the residual xylan content in pulps being stimulated by additional cellulase usage. The action of the latter strongly depends on the cellulose allomorphy confirming the impact of the pulp morphology. Hence, the combined application of both types of enzymes offers a high potential for upgrading pulps in order to produce a pure and high-value cellulose product. Copyright © 2017 Elsevier Inc. All rights reserved.

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