Science.gov

Sample records for adsorbed protein films

  1. Influence of fluoride-detergent combinations on the visco-elasticity of adsorbed salivary protein films.

    PubMed

    Veeregowda, Deepak H; van der Mei, Henny C; Busscher, Henk J; Sharma, Prashant K

    2011-02-01

    The visco-elasticity of salivary-protein films is related to mouthfeel, lubrication, biofilm formation, and protection against erosion and is influenced by the adsorption of toothpaste components. The thickness and the visco-elasticity of hydrated films (determined using a quartz crystal microbalance) of 2-h-old in vitro-adsorbed salivary-protein films were 43.5 nm and 9.4 MHz, respectively, whereas the dehydrated thickness, measured using X-ray photoelectron spectroscopy, was 2.4 nm. Treatment with toothpaste slurries decreased the thickness of the film, depending on the fluoride-detergent combination involved. Secondary exposure to saliva resulted in a regained thickness of the film to a level similar to its original thickness; however, no association was found between the thickness of hydrated and dehydrated films, indicating differences in film structure. Treatment with stannous fluoride/sodium lauryl sulphate (SnF(2)/SLS)-containing toothpaste slurries yielded a strong, immediate two-fold increase in characteristic film frequency (f(c)) with respect to untreated films, indicating cross-linking in adsorbed salivary-protein films by Sn(2+) that was absent when SLS was replaced with sodium hexametaphosphate (NaHMP). Secondary exposure to saliva of films treated with SnF(2) caused a strong, six-fold increase in f(c) compared with primary salivary-protein films, regardless of whether SLS or NaHMP was the detergent. This suggests that ionized stannous is not directly available for cross-linking in combination with highly negatively charged NaHMP, but becomes slowly available after initial treatment to cause cross-linking during secondary exposure to saliva.

  2. Contribution of Adsorbed Protein Films to Nanoscopic Vibrations Exhibited by Bacteria Adhering through Ligand-Receptor Bonds.

    PubMed

    Song, Lei; Sjollema, Jelmer; Norde, Willem; Busscher, Henk J; van der Mei, Henny C

    2015-09-29

    Bacteria adhering to surfaces exhibit nanoscopic vibrations that depend on the viscoelasticity of the bond. The quantification of the nanoscopic vibrations of bacteria adhering to surfaces provides new opportunities to better understand the properties of the bond through which bacteria adhere and the mechanisms by which they resist detachment. Often, however, bacteria do not adhere to bare surfaces but to adsorbed protein films, on which adhesion involves highly specific ligand-receptor binding next to nonspecific DLVO interaction forces. Here we determine the contribution of adsorbed salivary protein and fibronectin films to vibrations exhibited by adhering streptococci and staphylococci, respectively. The streptococcal strain used has the ability to adhere to adsorbed salivary proteins films through antigen I/II ligand-receptor binding, while the staphylococcal strain used adheres to adsorbed fibronectin films through a proteinaceous ligand-receptor bond. In the absence of ligand-receptor binding, electrostatic interactions had a large impact on vibration amplitudes of adhering bacteria on glass. On an adsorbed salivary protein film, vibration amplitudes of adhering streptococci depended on the film softness as determined by QCM-D and were reduced after film fixation using glutaraldehyde. On a relatively stiff fibronectin film, cross-linking the film in glutaraldehyde hardly reduced its softness, and accordingly fibronectin film softness did not contribute to vibration amplitudes of adhering staphylococci. However, fixation of the staphylococcus-fibronectin bond further decreased vibration amplitudes, while fixation of the streptococcus bond hardly impacted vibration amplitudes. Summarizing, this study shows that both the softness of adsorbed protein films and the properties of the bond between an adhering bacterium and an adsorbed protein film play an important role in bacterial vibration amplitudes. These nanoscopic vibrations reflect the viscoelasticity of the

  3. Quantitative time-of-flight secondary ion mass spectrometry for the characterization of multicomponent adsorbed protein films

    NASA Astrophysics Data System (ADS)

    Wagner, M. S.; Shen, M.; Horbett, T. A.; Castner, David G.

    2003-01-01

    Time-of-flight secondary ion mass spectrometry (ToF-SIMS) is ideal for the characterization of adsorbed proteins due to its chemical specificity and surface sensitivity. We have employed ToF-SIMS and multivariate analysis to determine the surface composition of adsorbed protein films from binary mixtures, blood serum, and blood plasma. Good correlation between ToF-SIMS data and independent radiolabeling studies was achieved for binary mixtures, though these results depended on the substrate. Qualitative insight into the composition of the serum and plasma protein films was obtained via comparison to standard single protein film spectra. ToF-SIMS and multivariate analysis were able to measure the surface composition of multicomponent adsorbed protein films.

  4. ToF-SIMS and XPS Characterization of Protein Films Adsorbed onto Bare and Sodium Styrenesulfonate-Grafted Gold Substrates.

    PubMed

    Foster, Rami N; Harrison, Elisa T; Castner, David G

    2016-04-05

    The adsorption of single-component bovine serum albumin (BSA), bovine fibrinogen (Fgn), and bovine immunoglobulin G (IgG) films as well as multicomponent bovine plasma films onto bare and sodium styrenesulfonate (NaSS)-grafted gold substrates was characterized. The adsorption isotherms, measured via X-ray photoelectron spectroscopy, showed that at low solution concentrations all three single-component proteins adsorb with higher affinity onto gold surfaces compared to NaSS surfaces. However, at higher concentrations, NaSS surfaces adsorb the same or more total protein than gold surfaces. This may be because proteins that adsorb onto NaSS undergo structural rearrangements, resulting in a larger fraction of irreversibly adsorbed species over time. Still, with the possible exception of BSA adsorbed onto gold, neither surface appeared to have saturated at the highest protein solution concentration studied. Principal component (PC) analysis of amino acid mass fragments from time-of-flight secondary ion mass spectra distinguished between the same protein adsorbed onto NaSS and gold surfaces, suggesting that proteins adsorb differently on NaSS and gold surfaces. Explored further using peak ratios for buried/surface amino acids for each protein, we found that proteins denature more on NaSS surfaces than on gold surfaces. Also, using peak ratios for asymmetrically distributed amino acids, potential structural differences were postulated for BSA and IgG adsorbed onto NaSS and gold surfaces. PC modeling, used to track changes in plasma adsorption with time, suggests that plasma films on NaSS and Au surfaces become more Fgn-like with increasing adsorption time. However, the PC models included only three proteins, where plasma is composed of hundreds of proteins. Therefore, while both gold and NaSS appear to adsorb more Fgn with time, further study is required to confirm that this is representative of the final state of the plasma films.

  5. In situ investigations of Fe3+ induced complexation of adsorbed Mefp-1 protein film on iron substrate.

    PubMed

    Zhang, Fan; Sababi, Majid; Brinck, Tore; Persson, Dan; Pan, Jinshan; Claesson, Per M

    2013-08-15

    A range of in situ analytical techniques and theoretical calculations were applied to gain insights into the formation and properties of the Mefp-1 film on iron substrate, as well as the protein complexation with Fe(3+) ions. Adsorption kinetics of Mefp-1 and the complexation were investigated using QCM-D. The results suggest an initially fast adsorption, with the molecules oriented preferentially parallel to the surface, followed by a structural change within the film leading to molecules extending toward solution. Exposure to a diluted FeCl3 solution results in enhanced complexation within the adsorbed protein film, leading to water removal and film compaction. In situ Peak Force Tapping AFM was employed for determining morphology and nano-mechanical properties of the surface layer. The results, in agreement with the QCM-D observations, demonstrate that addition of Fe(3+) induces a transition from an extended and soft protein layer to a denser and stiffer one. Further, in situ ATR-FTIR and Confocal Raman Micro-spectroscopy (CRM) techniques were utilized to monitor compositional/structural changes in the surface layer due to addition of Fe(3+) ions. The spectroscopic analyses assisted by DFT calculations provide evidence for formation of tri-Fe(3+)/catechol complexes in the surface film, which is enhanced by Fe(3+) addition.

  6. Morphological changes in adsorbed protein films at the air-water interface subjected to large area variations, as observed by brewster angle microscopy.

    PubMed

    Xu, Rong; Dickinson, Eric; Murray, Brent S

    2007-04-24

    Adsorbed films of proteins at the air-water interface have been imaged using Brewster angle microscopy (BAM). The proteins beta-lactoglobulin (beta-L) and ovalbumin (OA) were studied at a range of protein concentrations and surface ages at 25.0 degrees C and two pH values (7 and 5) in a Langmuir trough. The adsorbed films were periodically subjected to compression and expansion cycles such that the film area was typically varied between 125% and 50% of the original film area. With beta-L on its own, no structural changes were observable at pH 7. When a low-area fraction (less than 0.01%) of 20 mum polystyrene latex particles was spread at the interface before adsorption of beta-L, the particles became randomly distributed throughout the interface, but after protein adsorption and compression/expansion, the particles highlighted notable structural features not visible in their absence. Such features included the appearance of long (several hundred micrometers or more) folds and cracks in the films, generally oriented at right angles to the direction of compression, and also aggregates of protein and/or particles. Such structuring was more visible the longer the film was aged or at higher initial protein concentrations for shorter adsorption times. At pH 5, close to the isoelectric pH of beta-L, such features were just noticeable in the absence of particles but were much more pronounced than at pH 7 in the presence of particles. Similar experiments with OA revealed even more pronounced structural features, both in the absence and presence of particles, particularly at pH 5 (close to the isoelectric pH of OA also), producing striking stripelike and meshlike domains. Changes in the dilatational elasticity of the films could be correlated with the variations in the structural integrity of the films as observed via BAM. The results indicate that interfacial area changes of this type, typical of those that occur in food colloid processing, will lead to highly

  7. Conformational changes of adsorbed proteins

    NASA Astrophysics Data System (ADS)

    Allen, Scott

    2005-03-01

    The adsorption of bovine serum albumin (BSA) and pepsin to gold surfaces has been studied using surface plasmon resonance (SPR). Proteins are adsorbed from solution onto a gold surface and changes in the conformation of the adsorbed proteins are induced by changing the buffer solution. We selected pH and ionic strength values for the buffer solutions that are known from our circular dichroism measurements to cause conformational changes of the proteins in bulk solution. We find that for both BSA and pepsin the changes in conformation are impeded by the interaction of the protein with the gold surface.

  8. Conformation Distributions in Adsorbed Proteins.

    NASA Astrophysics Data System (ADS)

    Meuse, Curtis W.; Hubbard, Joseph B.; Vrettos, John S.; Smith, Jackson R.; Cicerone, Marcus T.

    2007-03-01

    While the structural basis of protein function is well understood in the biopharmaceutical and biotechnology industries, few methods for the characterization and comparison of protein conformation distributions are available. New methods capable of measuring the stability of protein conformations and the integrity of protein-protein, protein-ligand and protein-surface interactions both in solution and on surfaces are needed to help the development of protein-based products. We are developing infrared spectroscopy methods for the characterization and comparison of molecular conformation distributions in monolayers and in solutions. We have extracted an order parameter describing the orientational and conformational variations of protein functional groups around the average molecular values from a single polarized spectrum. We will discuss the development of these methods and compare them to amide hydrogen/deuterium exchange methods for albumin in solution and on different polymer surfaces to show that our order parameter is related to protein stability.

  9. A Multi-technique Characterization of Adsorbed Protein Films: Orientation and Structure by ToF-SIMS, NEXAFS, SFG, and XPS

    NASA Astrophysics Data System (ADS)

    Baio, Joseph E.

    immobilization schemes. This protein contained both a hexahistidine tag and a cysteine residue, introduced at opposite ends of the HuLys Fv, for immobilization onto nitrilotriacetic acid (NTA) and maleimide oligo- (ethylene glycol) (MEG)-terminated substrates. The thiol group on the cysteine residue selectively binds to the MEG groups, while the his-tag selectively binds to the Ni-loaded NTA groups. XPS was used to monitor protein coverage on both surfaces by following the change in the nitrogen atomic %. The ToF-SIMS data provided a clear differentiation between the two samples due to the intensity differences of secondary ions originating from asymmetrically located amino acids in HuLys Fv. Indicating that the HuLys Fv fragment when adsorbed into the NTA and MEG substrates will be induced into two different orientations. On the NTA substrate the protein's binding site is accessible, while on the MEG substrate the binding site is oriented towards the surface. By taking advantage of the electron pathway through the heme group in cytochrome c (CytoC) electrochemists have built sensors based upon CytoC immobilized onto functionalized metal electrodes. When immobilized onto a charged surface, CytoC, with its distribution of lysine and glutamate residues around its surface, should orient and form a well-ordered protein film. Here a detailed examination of CytoC orientation when electrostatically immobilized onto both amine (NH 3+) and carboxyl (COO-) functionalized gold is presented. Again, protein coverage, on both surfaces, was monitored by the change in the atomic % N, as determined by XPS. ToF-SIMS data demonstrated a clear separation between the two samples based on the intensity differences of secondary ions stemming from amino acids located asymmetrically within CytoC, indicating opposite orientations of the protein on the two different surfaces. Spectral features within the in situ sum frequency generation vibrational spectra, acquired for the protein interacting with

  10. Hydraulic properties of adsorbed water films in unsaturated porous media

    SciTech Connect

    Tokunaga, Tetsu K.

    2009-03-01

    Adsorbed water films strongly influence residual water saturations and hydraulic conductivities in porous media at low saturations. Hydraulic properties of adsorbed water films in unsaturated porous media were investigated through combining Langmuir's film model with scaling analysis, without use of any adjustable parameters. Diffuse double layer influences are predicted to be important through the strong dependence of adsorbed water film thickness (f) on matric potential ({Psi}) and ion charge (z). Film thickness, film velocity, and unsaturated hydraulic conductivity are predicted to vary with z{sup -1}, z{sup -2}, and z{sup -3}, respectively. In monodisperse granular media, the characteristic grain size ({lambda}) controls film hydraulics through {lambda}{sup -1} scaling of (1) the perimeter length per unit cross sectional area over which films occur, (2) the critical matric potential ({Psi}{sub c}) below which films control flow, and (3) the magnitude of the unsaturated hydraulic conductivity when {Psi} < {Psi}{sub c}. While it is recognized that finer textured sediments have higher unsaturated hydraulic conductivities than coarser sands at intermediate {Psi}, the {lambda}{sup -1} scaling of hydraulic conductivity predicted here extends this understanding to very low saturations where all pores are drained. Extremely low unsaturated hydraulic conductivities are predicted under adsorbed film-controlled conditions (generally < 0.1 mm y{sup -1}). On flat surfaces, the film hydraulic diffusivity is shown to be constant (invariant with respect to {Psi}).

  11. Activity of alkaline phosphatase adsorbed and grafted on "polydopamine" films.

    PubMed

    Ball, Vincent

    2014-09-01

    The oxidation of dopamine in slightly basic solutions and in the presence of oxygen as an oxidant allows for the deposition of dopamine-eumelanin ("polydopamine") films on almost all kinds of materials allowing for an easy secondary functionalization. Molecules carrying nucleophilic groups like thiols and amines can be easily grafted on those films. Herein we show that alkaline phosphatase (ALP), as a model enzyme, adsorbs to "polydopamine" films and part of the adsorbed enzyme is rapidly desorbed in contact with Tris buffer. However a significant part of the enzyme remains irreversibly adsorbed and keeps some enzymatic activity for at least 2 weeks whereas ALP adsorbed on quartz slides is rapidly and quantitatively deactivated. In addition we estimated the Michaelis constant Km of the enzyme irreversibly bound to the "polydopamine" film. The Michaelis constant, and hence the affinity constant between paranitrophenol phosphate and ALP are almost identical between the enzyme bound on the film and the free enzyme in solution. Complementarily, it was found that "polydopamine" films display some phosphatase like catalytic activity.

  12. Anomalous conformational transitions in cytochrome C adsorbing to Langmuir-Blodgett films

    NASA Astrophysics Data System (ADS)

    Sankaranarayanan, Kamatchi; Nair, B. U.; Dhathathreyan, A.

    2013-05-01

    Helix to beta conformational transitions in proteins has attracted much attention due to their relevance to fibril formation which is implicated in many neurological diseases. This study reports on unusual conformational transition of cytochrome C adsorbing to hydrophilic surface containing pure cationic lipid and mixed Langmuir-Blodgett films (LB films) of cationic and neutral lipids. Evidence for conformational changes of the protein from its native helical state to beta sheet comes from Circular dichroic spectroscopy (CD spectroscopy). Analysis of these samples using High resolution TEM (HRTEM) shows a typical fibrillar pattern with each strand spacing of about 0.41 nm across which can be attributed to the repeat distance of interdigitated neighboring hydrogen-bonded ribbons in a beta sheet. Changes in contact angles of protein adsorbing to the LB films together with the increased mass uptake of water using quartz crystal microbalance (QCM) confirm the role of positive charges in the conformational transition. Dehydration of the protein resulting from the excess water entrainment in the polar planes of the cationic lipid in hydrophilic surface seems to trigger the refolding of the protein to beta sheet while it retains its native conformation in hydrophobic films. The results suggest that drastic conformational changes in CytC adsorbing to cationic lipids may be of significance in its role as a peripheral membrane protein.

  13. Design and characterization of complex protein films

    NASA Astrophysics Data System (ADS)

    Bui, Holt P.

    Once a biomaterial is implanted into biological system, a layer of protein is immediately deposited on the surface of that material. The newly formed protein film will dictate how the implanted material will interact with the surrounding biological environment and lead to either the acceptance or rejection of the biomaterial. One method to enhance performance involves the activation the surface of the biomaterial with one or more proteins to direct specific interactions with the host environment. The focus of my dissertation was to develop and characterize model biomaterials surfaces that are activated with one or more proteins to help understand how the protein films may affect biological processes and a biomaterial's performance. One model system consisted of a patterned film of two proteins on a gold surface. Characterization of this protein pattern indicated that patterning protein films with a focused ion beam produced protein patterns with high biological contrast and high spatial control. The second model protein film involved the adsorption of fibronectin on surfaces with different surface energies. The characterization of the adsorbed fibronectin films suggest that fibronectin adsorbed on a hydrophilic surface is in an orientation that projects hydrophilic amino acid residues towards surface of the protein and dehydration causes reorientation to project hydrophobic amino acids towards the surface. In contrast, fibronectin is adsorbed onto a hydrophobic surface in a manner that resulted in dehydration and denaturation during the adsorption process. The last model protein film studied in this work consisted of fibronectin patterned in a manner so that the film consisted of spatially controlled domains of fibronectin adsorbed onto a hydrophilic surface as well as a hydrophobic surface. Lateral characterization of this pattern demonstrated a difference in secondary structure of fibronectin adsorbed on the two domains with varying surface energies.

  14. Liquid 4He Adsorbed Films on Very Attractive Substrates

    NASA Astrophysics Data System (ADS)

    Urrutia, Ignacio; Szybisz, Leszek

    Adsorbed films of liquid 4He are analized, in the framework of Density functionals Theories (DF). In these systems, when the substrate becomes increasingly attractive, the thin films of 4He approaches the quasi-bidimensional limit. We study this strongly attractive substrate regime with two DF, the Orsay-Trento (OT) and a recent Hybrid proposal (Hyb), focusing in the energy behavior. It is showed that OT does not reproduce the correct limiting energy curve, and it implies that this functional could not provide reliable results for very strongly attractive substrates like Graphite (Gr). In other hand, with the Hyb DF, the correct energy behavior is found for the adsorption energy of 4He on Gr. These results show that OT should not be applied to quasi 2D (confinement) situations, and that Hyb DF provides a much more realistic description.

  15. Liquid 4He Adsorbed Films on Very Attractive Substrates

    NASA Astrophysics Data System (ADS)

    Urrutia, Ignacio; Szybisz, Leszek

    2006-09-01

    Adsorbed films of liquid 4He are analized, in the framework of Density Functional Theories (DF). In these systems, when the substrate becomes increasingly attractive, the thin films of 4He approaches the quasi-bidimensional limit. We study this strongly attractive substrate regime with two DF, the Orsay-Trento (OT) and a recent Hybrid proposal (Hyb), focusing in the energy behavior. It is showed that OT does not reproduce the correct limiting energy curve, and it implies that this functional could not provide reliable results for very strongly attractive substrates like Graphite (Gr). In other hand, with the Hyb DF, the correct energy behavior is found for the adsorption energy of 4He on Gr. These results show that OT should not be applied to quasi 2D (confinement) situations, and that Hyb DF provides a much more realistic description.

  16. Quantification of the Influence of Protein-Protein Interactions on Adsorbed Protein Structure and Bioactivity

    PubMed Central

    Wei, Yang; Thyparambil, Aby A.; Latour, Robert A.

    2013-01-01

    While protein-surface interactions have been widely studied, relatively little is understood at this time regarding how protein-surface interaction effects are influenced by protein-protein interactions and how these effects combine with the internal stability of a protein to influence its adsorbed-state structure and bioactivity. The objectives of this study were to develop a method to study these combined effects under widely varying protein-protein interaction conditions using hen egg-white lysozyme (HEWL) adsorbed on silica glass, poly(methyl methacrylate), and polyethylene as our model systems. In order to vary protein-protein interaction effects over a wide range, HEWL was first adsorbed to each surface type under widely varying protein solution concentrations for 2 h to saturate the surface, followed by immersion in pure buffer solution for 15 h to equilibrate the adsorbed protein layers in the absence of additionally adsorbing protein. Periodic measurements were made at selected time points of the areal density of the adsorbed protein layer as an indicator of the level of protein-protein interaction effects within the layer, and these values were then correlated with measurements of the adsorbed protein’s secondary structure and bioactivity. The results from these studies indicate that protein-protein interaction effects help stabilize the structure of HEWL adsorbed on silica glass, have little influence on the structural behavior of HEWL on HDPE, and actually serve to destabilize HEWL’s structure on PMMA. The bioactivity of HEWL on silica glass and HDPE was found to decrease in direct proportion to the degree of adsorption-induce protein unfolding. A direct correlation between bioactivity and the conformational state of adsorbed HEWL was less apparent on PMMA, thus suggesting that other factors influenced HEWL’s bioactivity on this surface, such as the accessibility of HEWL’s bioactive site being blocked by neighboring proteins or the surface

  17. Adsorbed Methane Film Properties in Nanoporous Carbon Monoliths

    NASA Astrophysics Data System (ADS)

    Soo, Yuchoong; Chada, Nagaraju; Beckner, Matthew; Romanos, Jimmy; Burress, Jacob; Pfeifer, Peter

    2013-03-01

    Carbon briquetting can increase methane storage capacity by reducing the useless void volume resulting in a better packing density. It is a robust and efficient space-filling form for an adsorbed natural gas vehicle storage tank. To optimize methane storage capacity, we studied three fabrication process parameters: carbon-to-binder ratio, compaction temperature, and pyrolysis temperature. We found that carbon-to-binder ratio and pyrolysis temperature both have large influences on monolith uptakes. We have been able to optimize these parameters for high methane storage. All monolith uptakes (up to 260 bar) were measured by a custom-built, volumetric, reservoir-type instrument. The saturated film density and the film thickness was determined using linear extrapolation on the high pressure excess adsorption isotherms. The saturated film density was also determined using the monolayer Ono-Kondo model. Film densities ranged from ca. 0.32 g/cm3 - 0.37 g/cm3.The Ono-Kondo model also determines the binding energy of methane. Binding energies were also determined from isosteric heats calculated from the Clausius-Clapeyron equation and compared with the Ono-Kondo model method. Binding energies from Ono-Kondo were ca. 7.8 kJ/mol - 10 kJ/mol. Work funded by California Energy Commission Contract #500-08-022.

  18. Experimental characterization of adsorbed protein orientation, conformation, and bioactivity

    PubMed Central

    Thyparambil, Aby A.; Wei, Yang; Latour, Robert A.

    2015-01-01

    Protein adsorption on material surfaces is a common phenomenon that is of critical importance in many biotechnological applications. The structure and function of adsorbed proteins are tightly interrelated and play a key role in the communication and interaction of the adsorbed proteins with the surrounding environment. Because the bioactive state of a protein on a surface is a function of the orientation, conformation, and accessibility of its bioactive site(s), the isolated determination of just one or two of these factors will typically not be sufficient to understand the structure–function relationships of the adsorbed layer. Rather a combination of methods is needed to address each of these factors in a synergistic manner to provide a complementary dataset to characterize and understand the bioactive state of adsorbed protein. Over the past several years, the authors have focused on the development of such a set of complementary methods to address this need. These methods include adsorbed-state circular dichroism spectropolarimetry to determine adsorption-induced changes in protein secondary structure, amino-acid labeling/mass spectrometry to assess adsorbed protein orientation and tertiary structure by monitoring adsorption-induced changes in residue solvent accessibility, and bioactivity assays to assess adsorption-induced changes in protein bioactivity. In this paper, the authors describe the methods that they have developed and/or adapted for each of these assays. The authors then provide an example of their application to characterize how adsorption-induced changes in protein structure influence the enzymatic activity of hen egg-white lysozyme on fused silica glass, high density polyethylene, and poly(methyl-methacrylate) as a set of model systems. PMID:25708632

  19. Pattern Recognition of Adsorbing HP Lattice Proteins

    NASA Astrophysics Data System (ADS)

    Wilson, Matthew S.; Shi, Guangjie; Wüst, Thomas; Landau, David P.; Schmid, Friederike

    2015-03-01

    Protein adsorption is relevant in fields ranging from medicine to industry, and the qualitative behavior exhibited by course-grained models could shed insight for further research in such fields. Our study on the selective adsorption of lattice proteins utilizes the Wang-Landau algorithm to simulate the Hydrophobic-Polar (H-P) model with an efficient set of Monte Carlo moves. Each substrate is modeled as a square pattern of 9 lattice sites which attract either H or P monomers, and are located on an otherwise neutral surface. The fully enumerated set of 102 unique surfaces is simulated with each protein sequence. A collection of 27-monomer sequences is used- each of which is non-degenerate and protein-like. Thermodynamic quantities such as the specific heat and free energy are calculated from the density of states, and are used to investigate the adsorption of lattice proteins on patterned substrates. Research supported by NSF.

  20. Fast and efficient protein purification using membrane adsorber systems.

    PubMed

    Suck, Kirstin; Walter, Johanna; Menzel, Frauke; Tappe, Alexander; Kasper, Cornelia; Naumann, Claudia; Zeidler, Robert; Scheper, Thomas

    2006-02-10

    The purification of proteins from complex cell culture samples is an essential step in proteomic research. Traditional chromatographic methods often require several steps resulting in time consuming and costly procedures. In contrast, protein purification via membrane adsorbers offers the advantage of fast and gentle but still effective isolation. In this work, we present a new method for purification of proteins from crude cell extracts via membrane adsorber based devices. This isolation procedure utilises the membranes favourable pore structure allowing high flow rates without causing high back pressure. Therefore, shear stress to fragile structures is avoided. In addition, mass transfer takes place through convection rather than diffusion, thus allowing very rapid separation processes. Based on this membrane adsorber technology the separation of two model proteins, human serum albumin (HSA) and immungluboline G (IgG) is shown. The isolation of human growth hormone (hGH) from chinese hamster ovary (CHO) cell culture supernatant was performed using a cation exchange membrane. The isolation of the enzyme penicillin acylase from the crude Escherichia coli supernatant was achieved using an anion exchange spin column within one step at a considerable purity. In summary, the membrane adsorber devices have proven to be suitable tools for the purification of proteins from different complex cell culture samples.

  1. The molecular mechanism of mediation of adsorbed serum proteins to endothelial cells adhesion and growth on biomaterials.

    PubMed

    Yang, Dayun; Lü, Xiaoying; Hong, Ying; Xi, Tingfei; Zhang, Deyuan

    2013-07-01

    To explore molecular mechanism of mediation of adsorbed proteins to cell adhesion and growth on biomaterials, this study examined endothelial cell adhesion, morphology and viability on bare and titanium nitride (TiN) coated nickel titanium (NiTi) alloys and chitosan film firstly, and then identified the type and amount of serum proteins adsorbed on the three surfaces by proteomic technology. Subsequently, the mediation role of the identified proteins to cell adhesion and growth was investigated with bioinformatics analyses, and further confirmed by a series of cellular and molecular biological experiments. Results showed that the type and amount of adsorbed serum proteins associated with cell adhesion and growth was obviously higher on the alloys than on the chitosan film, and these proteins mediated endothelial cell adhesion and growth on the alloys via four ways. First, proteins such as adiponectin in the adsorbed protein layer bound with cell surface receptors to generate signal transduction, which activated cell surface integrins through increasing intracellular calcium level. Another way, thrombospondin 1 in the adsorbed protein layer promoted TGF-β signaling pathway activation and enhanced integrins expression. The third, RGD sequence containing proteins such as fibronectin 1, vitronectin and thrombospondin 1 in the adsorbed protein layer bound with activated integrins to activate focal adhesion pathway, increased focal adhesion formation and actin cytoskeleton organization and mediated cell adhesion and spreading. In addition, the activated focal adhesion pathway promoted the expression of cell growth related genes and resulted in cell proliferation. The fourth route, coagulation factor II (F2) and fibronectin 1 in the adsorbed protein layer bound with cell surface F2 receptor and integrin, activated regulation of actin cytoskeleton pathway and regulated actin cytoskeleton organization.

  2. The Effect of monoglycerides on structural and topographical characteristics of adsorbed beta-casein films at the air-water interface.

    PubMed

    Fernández, Marta Cejudo; Sánchez, Cecilio Carrera; Rodríguez Niño, M Rosario; Rodríguez Patino, Juan M

    2006-02-01

    The effect of monoglycerides (monopalmitin and monoolein) on the structural and topographical characteristics of beta-casein adsorbed film at the air-water interface has been analyzed by means of surface pressure (pi)-area (A) isotherms and Brewster angle microscopy (BAM). At surface pressures lower than that for the beta-casein collapse (pi(c)(beta-casein)), attractive interactions between beta-casein and monoglycerides were observed. At higher surface pressures, the collapsed beta-casein is partially displaced from the interface by monoglycerides. However, beta-casein displacement by monoglycerides is not quantitative at the monoglyceride concentrations studied in this work. From the results derived from these experiments, we have concluded that interactions, miscibility, and displacement of proteins by monoglycerides in adsorbed mixed monolayers at the air-water interface depend on the particular protein-monoglyceride system, the interactions between film-forming components being higher for adsorbed than for spread films. The adsorbed films are more segregated than spread films, and both collapsed protein domains and monoglyceride domains in adsorbed films are smaller than for spread films.

  3. Ultra-Thin Optically Transparent Carbon Electrodes Produced from Layers of Adsorbed Proteins

    PubMed Central

    Alharthi, Sarah A.; Benavidez, Tomas E.; Garcia, Carlos D.

    2013-01-01

    This work describes a simple, versatile, and inexpensive procedure to prepare optically transparent carbon electrodes, using proteins as precursors. Upon adsorption, the protein-coated substrates were pyrolyzed under reductive conditions (5% H2) to form ultra-thin, conductive electrodes. Because proteins spontaneously adsorb to interfaces forming uniform layers, the proposed method does not require a precise control of the preparation conditions, specialized instrumentation, or expensive precursors. The resulting electrodes were characterized by a combination of electrochemical, optical, and spectroscopic means. As a proof-of-concept, the optically-transparent electrodes were also used as substrate for the development of an electrochemical glucose biosensor. The proposed films represent a convenient alternative to more sophisticated, and less available, carbon-based nanomaterials. Furthermore, these films could be formed on a variety of substrates, without classical limitations of size or shape. PMID:23421732

  4. Protein thin film machines.

    PubMed

    Federici, Stefania; Oliviero, Giulio; Hamad-Schifferli, Kimberly; Bergese, Paolo

    2010-12-01

    We report the first example of microcantilever beams that are reversibly driven by protein thin film machines fueled by cycling the salt concentration of the surrounding solution. We also show that upon the same salinity stimulus the drive can be completely reversed in its direction by introducing a surface coating ligand. Experimental results are throughout discussed within a general yet simple thermodynamic model.

  5. Influence of ionic strength changes on the structure of pre-adsorbed salivary films. A response of a natural multi-component layer.

    PubMed

    Macakova, Lubica; Yakubov, Gleb E; Plunkett, Mark A; Stokes, Jason R

    2010-05-01

    Salivary films coating oral surfaces are critically important for oral health. This study focuses on determining the underlying nature of this adsorbed film and how it responds to departures from physiological conditions due to changes in ionic strength. Under physiological conditions, it is found that pre-adsorbed in vitro salivary film on hydrophobic surfaces is present as a highly hydrated viscoelastic layer. We follow the evolution of this film in terms of its effective thickness, hydration and viscoelastic properties, as well as adsorbed mass of proteins, using complementary surface characterisation methods: a Surface Plasmon Resonance (SPR) and a Quartz Crystal Microbalance with Dissipation Monitoring (QCM-D). Our results support a heterogeneous model for the structure of the salivary film with an inner dense anchoring layer and an outer highly extended hydrated layer. Further swelling of the film was observed upon decreasing the salt concentration down to 1mM NaCl. However, upon exposure to deionised water, a collapse of the film occurs that was associated with the loss of water contained within the adsorbed layer. We suggest that the collapse in deionised water is driven by an onset of electrostatic attraction between different parts of the multi-component salivary film. It is anticipated that such changes could also occur when the oral cavity is exposed to food, beverage, oral care and pharmaceutical formulations where drastic changes to the structural integrity of the film is likely to have implications on oral health, sensory perception and product performance.

  6. Interaction of magnetic nanoparticles with phospholipid films adsorbed at a liquid/liquid interface.

    PubMed

    Cámara, C I; Monzón, L M A; Coey, J M D; Yudi, L M

    2015-01-07

    The interaction of Co hexagonal magnetic nanoparticles (MNPs) with distearoyl phosphatidyl glycerol (DSPG) and distearoyl phosphatidic acid (DSPA) films adsorbed at a water/1,2-dichloroethane interface is studied employing cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), capacity curves and interfacial pressure-area isotherms. DSPA and DSPG adsorb at the interface forming homogenous films and producing a blocking effect on the transfer process of tetraethyl ammonium (TEA(+)), used as a probe cation. In the presence of Co NPs this effect is reversed and the reversible transfer process for TEA(+) is reestablished, to a greater or lesser extent depending on the structuration of the film. Co-DSPA hybrid films have a homogeneous structure while Co-DSPG films present different domains. Moreover, the presence of Co on DSPA film modifies the partition coefficient of the organic electrolyte into the hydrocarbon layer.

  7. Adsorption and separation of proteins by collagen fiber adsorbent.

    PubMed

    Li, Juan; Liao, Xue-pin; Zhang, Qi-xian; Shi, Bi

    2013-06-01

    The separation of proteins is a key step in biomedical and pharmaceutical industries. In the present investigation, the collagen fiber adsorbent (CFA) was exploited as column packing material to separate proteins. Bovine serum albumin (BSA), bovine hemoglobin (Hb) and lysozyme (LYS) that have different isoelectric points (pIs) were selected as model proteins to investigate the separation ability of CFA to proteins. In batch adsorption, the adsorption behaviors of these proteins on CFA under different pHs and ionic strengths indicated that the electrostatic interaction plays a predominant role in the adsorption of proteins on CFA. CFA exhibited high adsorption capacity to Hb and LYS. In column separation, the proteins were completely separated by adjusting pH and ionic strength of the eluent. The increase of flow rate could reduce the separation time with no influence on the recovery of protein in the experimental range. The protein recovery was higher than 90% even when the CFA column was re-used for 4 times in separation of BSA and LYS, and the retention time of BSA or LYS was almost constant during the repeated applications. In addition, as a practical application, LYS was successfully separated from chicken egg white powder by CFA column.

  8. Structures of multidomain proteins adsorbed on hydrophobic interaction chromatography surfaces.

    PubMed

    Gospodarek, Adrian M; Sun, Weitong; O'Connell, John P; Fernandez, Erik J

    2014-12-05

    In hydrophobic interaction chromatography (HIC), interactions between buried hydrophobic residues and HIC surfaces can cause conformational changes that interfere with separations and cause yield losses. This paper extends our previous investigations of protein unfolding in HIC chromatography by identifying protein structures on HIC surfaces under denaturing conditions and relating them to solution behavior. The thermal unfolding of three model multidomain proteins on three HIC surfaces of differing hydrophobicities was investigated with hydrogen exchange mass spectrometry (HXMS). The data were analyzed to obtain unfolding rates and Gibbs free energies for unfolding of adsorbed proteins. The melting temperatures of the proteins were lowered, but by different amounts, on the different surfaces. In addition, the structures of the proteins on the chromatographic surfaces were similar to the partially unfolded structures produced in the absence of a surface by temperature as well as by chemical denaturants. Finally, it was found that patterns of residue exposure to solvent on different surfaces at different temperatures can be largely superimposed. These findings suggest that protein unfolding on various HIC surfaces might be quantitatively related to protein unfolding in solution and that details of surface unfolding behavior might be generalized.

  9. Heat capacity measurements of atoms and molecules adsorbed on evaporated metal films

    SciTech Connect

    Kenny, T.W.

    1989-05-01

    Investigations of the properties of absorbed monolayers have received great experimental and theoretical attention recently, both because of the importance of surface processes in practical applications such as catalysis, and the importance of such systems to the understanding of the fundamentals of thermodynamics in two dimensions. We have adapted the composite bolometer technology to the construction of microcalorimeters. For these calorimeters, the adsorption substrate is an evaporated film deposited on one surface of an optically polished sapphire wafer. This approach has allowed us to make the first measurements of the heat capacity of submonolayer films of /sup 4/He adsorbed on metallic films. In contrast to measurements of /sup 4/He adsorbed on all other insulating substrates, we have shown that /sup 4/He on silver films occupies a two-dimensional gas phase over a broad range of coverages and temperatures. Our apparatus has been used to study the heat capacity of Indium flakes. CO multilayers, /sup 4/He adsorbed on sapphire and on Ag films and H/sub 2/ adsorbed on Ag films. The results are compared with appropriate theories. 68 refs., 19 figs.

  10. Unusual Morphologies of Poly(vinyl alcohol) Thin Films Adsorbed on Poly(dimethylsiloxane) Substrates.

    PubMed

    Karki, Akchheta; Nguyen, Lien; Sharma, Bhanushee; Yan, Yan; Chen, Wei

    2016-04-05

    Adsorption of poly(vinyl alcohol) (PVOH), 99% and 88% hydrolyzed poly(vinyl acetate), to poly(dimethylsiloxane) (PDMS) substrates was studied. The substrates were prepared by covalently attaching linear PDMS polymers of 2, 9, 17, 49, and 116 kDa onto silicon wafers. As the PDMS molecular weight/thickness increases, the adsorbed PVOH thin films progressively transition from continuous to discontinuous morphologies, including honeycomb and fractal/droplet. The structures are the result of thin film dewetting that occurs upon exposure to air. The PVOH film thickness does not vary significantly on these PDMS substrates, implicating the PDMS thickness as the cause for the morphology differences. The adsorbed PVOH thin films are less stable and have a stronger tendency to dewet on thicker, more liquid-like PDMS layers. When PVOH(99%) and PVOH(88%) thin films are compared, fractal and droplet morphologies are observed on high molecular weight PDMS substrates, respectively. The formation of the unique fractal features in the PVOH(99%) thin films as well as other crystalline and semicrystalline thin films is most likely driven by crystallization during the dehydration process in a diffusion-limited aggregation fashion. The only significant enhancement in hydrophilicity via PVOH adsorption was obtained on PDMS(2k), which is completely covered with a PVOH thin film. To mimic the lower receding contact angle and less liquid-like character of the PDMS(2k) substrate, light plasma treatment of the higher molecular weight PDMS substrates was carried out. On the treated PDMS substrates, the adsorbed PVOH thin films are in the more continuous honeycomb morphology, giving rise to significantly enhanced wettability. Furthermore, hydrophobic recovery of the hydrophilized PDMS substrates was not observed during a 1 week period. Thus, light plasma oxidation and subsequent PVOH adsorption can be utilized as a means to effectively hydrophilize conventional PDMS substrates. This study

  11. Long-term storage of surface-adsorbed protein machines.

    PubMed

    Albet-Torres, Nuria; Månsson, Alf

    2011-06-07

    The effective and simple long-term storage of complex functional proteins is critical in achieving commercially viable biosensors. This issue is particularly challenging in recently proposed types of nanobiosensors, where molecular-motor-driven transportation substitutes microfluidics and forms the basis for novel detection schemes. Importantly, therefore, we here describe that delicate heavy meromyosin (HMM)-based nanodevices (HMM motor fragments adsorbed to silanized surfaces and actin bound to HMM) fully maintain their function when stored at -20 °C for more than a month. The mechanisms for the excellent preservation of acto-HMM motor function upon repeated freeze-thaw cycles are discussed. The results are important to the future commercial implementation of motor-based nanodevices and are of more general value to the long-term storage of any protein-based bionanodevice.

  12. Spectrophotochemical and electrochemical characterization of perylene derivatives adsorbed on nanoporous metaloxide films

    NASA Astrophysics Data System (ADS)

    Kus, M.; Demic, S.; Zafer, C.; Saygili, G.; Bilgili, H.; Icli, S.

    2007-03-01

    Electrochemistry of perylene imide and anhydride derivatives adsorbed on semiconductor TiO{2} (NT) and insulator Al{2}O{3} (NA) metal oxide films were presented. Adsorption rates on metal oxide surface are observed to be strongly depending on molecular structure. Monoanhydride-monoimide derivatives show two reversible reductions in solution and one reversible reduction in films. Color change from red to blue and violet is observed indicating the formation of monoanion and dianion radicals. Spectroelectrochemical measurements support this interpretation. The color reversal is quite stable in NA films in comparison with NT films. This paper has been presented at “ECHOS06”, Paris, 28 30 juin 2006.

  13. In situ ATR and DRIFTS studies of the nature of adsorbed CO₂ on tetraethylenepentamine films.

    PubMed

    Wilfong, Walter Christopher; Srikanth, Chakravartula S; Chuang, Steven S C

    2014-08-27

    CO2 adsorption/desorption onto/from tetraethylenepentamine (TEPA) films of 4, 10, and 20 μm thicknesses were studied by in situ attenuated total reflectance (ATR) and diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) techniques under transient conditions. Molar absorption coefficients for adsorbed CO2 were used to determine the CO2 capture capacities and amine efficiencies (CO2/N) of the films in the DRIFTS system. Adsorption of CO2 onto surface and bulk NH2 groups of the 4 μm film produced weakly adsorbed CO2, which can be desorbed at 50 °C by reducing the CO2 partial pressure. These weakly adsorbed CO2 exhibit low ammonium ion intensities and could be in the form of ammonium-carbamate ion pairs and zwitterions. Increasing the film thickness enhanced the surface amine-amine interactions, resulting in strongly adsorbed ion pairs and zwitterions associated with NH and NH2 groups of neighboring amines. These adsorbed species may form an interconnected surface network, which slowed CO2 gas diffusion into and diminished access of the bulk amine groups (or amine efficiency) of the 20 μm film by a minimum of 65%. Desorption of strongly adsorbed CO2 comprising the surface network could occur via dissociation of NH3(+)/NH2(+)···NH2/NH ionic hydrogen bonds beginning from 60 to 80 °C, followed by decomposition of NHCOO(-)/NCOO(-) at 100 °C. These results suggest that faster CO2 diffusion and adsorption/desorption kinetics could be achieved by thinner layers of liquid or immobilized amines.

  14. Stability and interface properties of thin cellulose ester films adsorbed from acetone and ethyl acetate solutions.

    PubMed

    Amim, Jorge; Kosaka, Priscila M; Petri, Denise F S; Maia, Francisco C B; Miranda, Paulo B

    2009-04-15

    Stability and interface properties of cellulose acetate propionate (CAP) and cellulose acetate butyrate (CAB) films adsorbed from acetone or ethyl acetate onto Si wafers have been investigated by means of contact angle measurements and atomic force microscopy (AFM). Surface energy (gamma(S)(total)) values determined for CAP adsorbed from acetone are larger than those from ethyl acetate. In the case of CAB films adsorbed from ethyl acetate and acetone were similar. Dewetting was observed by AFM only for CAP films prepared from ethyl acetate. Positive values of effective Hamaker constant (A(eff)) were found only for CAP prepared from ethyl acetate, corroborating with dewetting phenomena observed by AFM. On the contrary, negative values of A(eff) were determined for CAP and CAB prepared from acetone and for CAB prepared from ethyl acetate, corroborating with experimental observations. Sum frequency generation (SFG) vibrational spectra indicated that CAP and CAB films prepared from ethyl acetate present more alkyl groups oriented perpendicularly to the polymer-air interface than those films prepared from acetone. Such preferential orientation corroborates with macroscopic contact angle measurements. Moreover, SFG spectra showed that acetone binds strongly to Si wafers, creating a new surface for CAP and CAB films.

  15. Miscibility of sodium chloride and sodium dodecyl sulfate in the adsorbed film and aggregate.

    PubMed

    Iyota, Hidemi; Krastev, Rumen

    2009-04-01

    The adsorption, micelle formation, and salting out of sodium dodecyl sulfate in the presence of sodium chloride were studied from the viewpoint of their mixed adsorption and aggregate formation. The surface tension of aqueous solutions of a sodium chloride-sodium dodecyl sulfate mixture was measured as a function of the total molality and composition of the mixture. Phase diagrams of adsorption and aggregate formation were obtained by applying thermodynamic equations to the surface tension. Judging from the phase diagrams, sodium chloride and sodium dodecyl sulfate are miscible in the adsorbed film at very large composition of sodium chloride and in the salted-out crystalline particle, while they are immiscible in the micelle. The miscibilities in the adsorbed film, micelle, and crystalline particle increase in the following order: particle > adsorbed film > micelle. The difference in miscibility among the oriented states was ascribed to the difference in geometry between the adsorbed film and micelle and to the interaction between bilayer surfaces in the particle.

  16. Adsorbate-induced demagnetization and restructuring of ultrathin magnetic films: CO chemisorbed on γ-Fe/Cu(100)

    NASA Astrophysics Data System (ADS)

    Spišák, D.; Hafner, J.

    2001-09-01

    First-principles local-spin-density (LSD) investigations of the structural, magnetic, and electronic properties of clean and CO-adsorbed ultrathin γ-iron films epitaxially grown on Cu(100) surfaces demonstrate that both the geometrical and the magnetic structures of the films are profoundly modified by the adsorption of CO. The enhanced magnetic moments of the top-layer atoms are strongly quenched by the presence of the adsorbate. Due to the pronounced magnetovolume effect, this leads also to a correlated change in the interlayer relaxations. Strikingly, the adsorbate-induced demagnetization is primarily limited to those surface atoms directly bonded to the adsorbate. This leads to the formation of an in-plane magnetic pattern in a partially adsorbate-covered film. The comparison of the calculated vibrational eigenfrequencies of the CO adsorbate with experiment confirms the picture based on the LSD calculations.

  17. Smooth deuterated cellulose films for the visualisation of adsorbed bio-macromolecules

    PubMed Central

    Su, Jielong; Raghuwanshi, Vikram S.; Raverty, Warwick; Garvey, Christopher J.; Holden, Peter J.; Gillon, Marie; Holt, Stephen A.; Tabor, Rico; Batchelor, Warren; Garnier, Gil

    2016-01-01

    Novel thin and smooth deuterated cellulose films were synthesised to visualize adsorbed bio-macromolecules using contrast variation neutron reflectivity (NR) measurements. Incorporation of varying degrees of deuteration into cellulose was achieved by growing Gluconacetobacter xylinus in deuterated glycerol as carbon source dissolved in growth media containing D2O. The derivative of deuterated cellulose was prepared by trimethylsilylation(TMS) in ionic liquid(1-butyl-3-methylimidazolium chloride). The TMS derivative was dissolved in toluene for thin film preparation by spin-coating. The resulting film was regenerated into deuterated cellulose by exposure to acidic vapour. A common enzyme, horseradish peroxidase (HRP), was adsorbed from solution onto the deuterated cellulose films and visualized by NR. The scattering length density contrast of the deuterated cellulose enabled accurate visualization and quantification of the adsorbed HRP, which would have been impossible to achieve with non-deuterated cellulose. The procedure described enables preparing deuterated cellulose films that allows differentiation of cellulose and non-deuterated bio-macromolecules using NR. PMID:27796332

  18. Smooth deuterated cellulose films for the visualisation of adsorbed bio-macromolecules.

    PubMed

    Su, Jielong; Raghuwanshi, Vikram S; Raverty, Warwick; Garvey, Christopher J; Holden, Peter J; Gillon, Marie; Holt, Stephen A; Tabor, Rico; Batchelor, Warren; Garnier, Gil

    2016-10-31

    Novel thin and smooth deuterated cellulose films were synthesised to visualize adsorbed bio-macromolecules using contrast variation neutron reflectivity (NR) measurements. Incorporation of varying degrees of deuteration into cellulose was achieved by growing Gluconacetobacter xylinus in deuterated glycerol as carbon source dissolved in growth media containing D2O. The derivative of deuterated cellulose was prepared by trimethylsilylation(TMS) in ionic liquid(1-butyl-3-methylimidazolium chloride). The TMS derivative was dissolved in toluene for thin film preparation by spin-coating. The resulting film was regenerated into deuterated cellulose by exposure to acidic vapour. A common enzyme, horseradish peroxidase (HRP), was adsorbed from solution onto the deuterated cellulose films and visualized by NR. The scattering length density contrast of the deuterated cellulose enabled accurate visualization and quantification of the adsorbed HRP, which would have been impossible to achieve with non-deuterated cellulose. The procedure described enables preparing deuterated cellulose films that allows differentiation of cellulose and non-deuterated bio-macromolecules using NR.

  19. Smooth deuterated cellulose films for the visualisation of adsorbed bio-macromolecules

    NASA Astrophysics Data System (ADS)

    Su, Jielong; Raghuwanshi, Vikram S.; Raverty, Warwick; Garvey, Christopher J.; Holden, Peter J.; Gillon, Marie; Holt, Stephen A.; Tabor, Rico; Batchelor, Warren; Garnier, Gil

    2016-10-01

    Novel thin and smooth deuterated cellulose films were synthesised to visualize adsorbed bio-macromolecules using contrast variation neutron reflectivity (NR) measurements. Incorporation of varying degrees of deuteration into cellulose was achieved by growing Gluconacetobacter xylinus in deuterated glycerol as carbon source dissolved in growth media containing D2O. The derivative of deuterated cellulose was prepared by trimethylsilylation(TMS) in ionic liquid(1-butyl-3-methylimidazolium chloride). The TMS derivative was dissolved in toluene for thin film preparation by spin-coating. The resulting film was regenerated into deuterated cellulose by exposure to acidic vapour. A common enzyme, horseradish peroxidase (HRP), was adsorbed from solution onto the deuterated cellulose films and visualized by NR. The scattering length density contrast of the deuterated cellulose enabled accurate visualization and quantification of the adsorbed HRP, which would have been impossible to achieve with non-deuterated cellulose. The procedure described enables preparing deuterated cellulose films that allows differentiation of cellulose and non-deuterated bio-macromolecules using NR.

  20. Zeta potential, contact angles, and AFM imaging of protein conformation adsorbed on hybrid nanocomposite surfaces.

    PubMed

    Pinho, Ana C; Piedade, Ana P

    2013-08-28

    The sputtering deposition of gold (Au) and poly(tetrafluoroethylene) (PTFE) was used to prepare a nanocomposite hybrid thin film suitable for protein adsorption while maintaining the native conformation of the biological material. The monolithic PTFE and the nanocomposite PTFE/Au thin films, with Au content up to 1 at %, were co-deposited by r.f. magnetron sputtering using argon as a discharge gas and deposited onto 316L stainless steel substrates, the most commonly used steel in biomaterials. The deposited thin films, before and after bovine serum albumin (BSA) adsorption, were thoroughly characterized with special emphasis on the surface properties/characteristics by atomic force microscopy (AFM), zeta potential, and static and dynamic contact angle measurements, in order to assess the relationship between structure and conformational changes. The influence of a pre-adsorbed peptide (RGD) was also evaluated. The nanotopographic and chemical changes induced by the presence of gold in the nanocomposite thin films enable RGD bonding, which is critical for the maintenance of the BSA native conformation after adsorption.

  1. Photoelectron decay kinetics of cubic silver chloride microcrystal film adsorbing plentiful dye excited by laser

    NASA Astrophysics Data System (ADS)

    Zhang, Rongxiang; Zhang, Jixian; Lai, Weidong; Hu, Yanxia; Dai, Xiuhong; Han, Li; Li, Xiaowei

    2007-12-01

    There will be large numbers of carriers coming into being in the interior of silver chloride microcrystals when illumination acts on it. Microwave absorption and dielectric spectrum detection technology with high temporal resolution (1ns) can detect instantaneous decay process of photoelectrons. In this work, the photoelectron decay action of spectral sensitized silver chloride emulsion is measured by microwave absorption and dielectric spectrum detection technology. By analyzing the measured results, it is found that when plentiful dye adsorb on silver chloride microcrystals film, the photoelectron decay of silver chloride emulsion becomes faster than that of pure emulsion. However it is not that the more the dye is adsorbed, the faster the photoelectron decay will be. When the adsorbed dye reaches a certain level, the photoelectron decay becomes slower than the fastest instance. Combining with photoelectron decay kinetics theory it is found that the above results are induced by two kinds of effect from dye adsorption.

  2. Characterization of cross-linked cellulosic ion-exchange adsorbents: 2. Protein sorption and transport.

    PubMed

    Angelo, James M; Cvetkovic, Aleksandar; Gantier, Rene; Lenhoff, Abraham M

    2016-03-18

    Adsorption behavior in the HyperCel family of cellulosic ion-exchange materials (Pall Corporation) was characterized using methods to assess, quantitatively and qualitatively, the dynamics of protein uptake as well as static adsorption as a function of ionic strength and protein concentration using several model proteins. The three exchangers studied all presented relatively high adsorptive capacities under low ionic strength conditions, comparable to commercially available resins containing polymer functionalization aimed at increasing that particular characteristic. The strong cation- and anion-exchange moieties showed higher sensitivity to increasing salt concentrations, but protein affinity on the salt-tolerant STAR AX HyperCel exchanger remained strong at ionic strengths normally used in downstream processing to elute material fully during ion-exchange chromatography. Very high uptake rates were observed in both batch kinetics experiments and time-series confocal laser scanning microscopy, suggesting low intraparticle transport resistances relative to external film resistance, even at higher bulk protein concentrations where the opposite is typically observed. Electron microscopy imaging of protein adsorbed phases provided additional insight into particle structure that could not be resolved in previous work on the bare resins.

  3. Film morphology and orientation of amino silicone adsorbed onto cellulose substrate

    NASA Astrophysics Data System (ADS)

    Xu, Yingjun; Yin, Hong; Yuan, Shenfeng; Chen, Zhirong

    2009-07-01

    A series of amino silicones with different amino values were synthesized and adsorbed onto surfaces of cotton fibers and cellulose substrates. The film morphology, hydrophobic properties and surface composition of the silicones are investigated and characterized by field emission scanning electron microscope (FESEM), atomic force microscope (AFM), contact angle measurement, X-ray photoelectron spectroscopy (XPS) and attenuated total reflectance infrared (ATR-IR). The results of the experiments indicate that the amino silicone can form a hydrophobic film on both cotton fibers and cellulose substrates and reduce the surface roughness significantly. Furthermore, the roughness becomes smaller with an increase in the amino value. All these results suggest that the orientation of amino silicone molecule is with the amino functional groups of amino silicone molecule adsorbed onto the cellulose interface while the main polymer chains and the hydrophobic Si-CH 3 groups extend toward the air.

  4. Aggregate formation of eosin-Y adsorbed on nanocrystalline TiO2 films

    NASA Astrophysics Data System (ADS)

    Yaguchi, Kaori; Furube, Akihiro; Katoh, Ryuzi

    2012-11-01

    We have studied the adsorption of eosin-Y on nanocrystalline TiO2 films with two different solvents namely acetonitrile (ACN) and ethanol (EtOH). A Langmuir-type adsorption isotherm was observed with ACN. In contrast, a Freundlich-type adsorption isotherm was observed with EtOH, suggesting that EtOH molecules co-adsorbed on TiO2 surface. Absorption spectra of the dye adsorbed films clearly show aggregate formation at high concentrations of dye in the solutions. From the analysis of the spectra, we conclude that head-to-tail type aggregates are observed with ACN, whereas various types of aggregates, including H-type and head-to-tail type aggregates, are observed with EtOH.

  5. Determination of anisotropic optical constants and surface coverage of molecular films using polarized visible ATR spectroscopy. Application to adsorbed cytochrome c films.

    PubMed

    Runge, Anne F; Rasmussen, Nicole C; Saavedra, S Scott; Mendes, Sergio B

    2005-01-13

    This article describes a method to determine the anisotropic optical constants and surface coverage of molecular films using polarized attenuated total reflectance (ATR) absorbance measurements. We have extended the transfer-matrix formalism to describe birefringent and dichroic films in ATR geometries and have combined it with an iterative numerical procedure to determine the anisotropic values of both the real (n) and imaginary (k) parts of the complex refractive index of the film under investigation. Anisotropic values of the imaginary part of the refractive index (k) allow for the determination of the surface coverage and one order parameter of the film. To illustrate this approach, we have used cytochrome c (cyt c) protein films adsorbed to glass and indium tin oxide (ITO) surfaces. Experimental results show that cyt c films on these surfaces, which were formed under identical conditions, have significant differences in their surface coverages (11.2 +/- 0.4 pmol/cm(2) on glass and 21.7 +/- 0.9 pmol/cm(2) on ITO); however, their order parameters are similar (0.30 +/- 0.02 on glass and 0.36 +/- 0.04 on ITO).

  6. Miscibility and interaction between 1-alkanol and short-chain phosphocholine in the adsorbed film and micelles.

    PubMed

    Takajo, Yuichi; Matsuki, Hitoshi; Kaneshina, Shoji; Aratono, Makoto; Yamanaka, Michio

    2007-09-01

    The miscibility and interaction of 1-hexanol (C6OH) and 1-heptanol (C7OH) with 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC) in the adsorbed films and micelles were investigated by measuring the surface tension of aqueous C6OH-DHPC and aqueous C7OH-DHPC solutions. The surface density, the mean molecular area, the composition of the adsorbed film, and the excess Gibbs energy of adsorption g(H,E), were estimated. Further, the critical micelle concentration of the mixtures was determined from the surface tension versus molality curves; the micellar composition was calculated. The miscibility of the 1-alkanols and DHPC molecules in the adsorbed film and micelles was examined using the phase diagram of adsorption (PDA) and that of micellization (PDM). The PDA and the composition dependence of g(H,E) indicated the non-ideal mixing of the 1-alkanols and DHPC molecules due to the attractive interaction between the molecules in the adsorbed film, while the PDM indicated that the 1-alkanol molecules were not incorporated in the micelles within DHPC rich region. The dependence of the mean molecular area of the mixtures on the surface composition suggested that the packing property of the adsorbed film depends on the chain length of 1-alkanol: C6OH expands the DHPC adsorbed film more than C7OH.

  7. Multi-technique Characterization of Adsorbed Peptide and Protein Orientation: LK310 and Protein G B1

    SciTech Connect

    Baio, J.; Weidner, T; Samuel, N; McCrea, K; Baugh, L; Stayton, P; Castner, D

    2010-01-01

    The ability to orient biologically active proteins on surfaces is a major challenge in the design, construction, and successful deployment of many medical technologies. As methods to orient biomolecules are developed, it is also essential to develop techniques that can accurately determine the orientation and structure of these materials. In this study, two model protein and peptide systems are presented to highlight the strengths of three surface analysis techniques for characterizing protein films: time-of-flight secondary-ion mass spectrometry (ToF-SIMS), sum-frequency generation (SFG) vibrational spectroscopy, and near-edge x-ray absorption fine structure (NEXAFS) spectroscopy. First, the orientation of Protein G B1, a rigid 6 kDa domain covalently attached to a maleimide-functionalized self-assembled monolayer, was examined using ToF-SIMS. Although the thickness of the Protein G layer was similar to the ToF-SIMS sampling depth, orientation of Protein G was successfully determined by analyzing the C{sub 2}H{sub 5}S{sup +} intensity, a secondary-ion derived from a methionine residue located at one end of the protein. Next, the secondary structure of a 13-mer leucine-lysine peptide (LK{sub 310}) adsorbed onto hydrophilic quartz and hydrophobic fluorocarbon surfaces was examined. SFG spectra indicated that the peptide's lysine side chains were ordered on the quartz surface, while the peptide's leucine side chains were ordered on the fluorocarbon surface. NEXAFS results provided complementary information about the structure of the LK{sub 310} film and the orientations of amide bonds within the LK{sub 310} peptide.

  8. Miscibility of Hydrocarbon and Fluorocarbon Surfactants in Adsorbed Film and Micelle.

    PubMed

    Villeneuve, Masumi; Nomura, Teruko; Matsuki, Hitoshi; Kaneshina, Shoji; Aratono, Makoto

    2001-02-01

    We investigated the miscibility of nonionic hydrocarbon and fluorocarbon surfactants in the adsorbed film and the micelle by surface tension measurements of the aqueous solution. The combination of tetraethyleneglycol monodecyl ether (C10E4) and tetraethyleneglycol mono-1,1,7-trihydrododecafluoroheptyl ether (FC7E4) was chosen because they have the same hydrophilic groups and about the same surface activity. The extent of nonideal mixing was estimated quantitatively in terms of the excess Gibbs energy in the adsorbed film g(H,E) and that in the micelle g(M,E). The excess area per adsorbed molecule, A(H,E), was also evaluated and discussed. The ionic hydrocarbon and fluorocarbon mixed surfactant systems, lithium dodecyl sulfate (LiDS)/lithium perfluorooctane sulfonate (LiFOS) and lithium tetradecyl sulfate (LiTS)/LiFOS systems are also investigated from the standpoint of excess Gibbs energy and excess area. It is also clearly shown that the regular solution approach does not fit in the systems that contain ionic species. Copyright 2001 Academic Press.

  9. AC microcalorimetry of adsorbates on evaporated metal films: Orientational ordering of H{sub 2} multilayers

    SciTech Connect

    Phelps, R.B.

    1991-11-01

    We have improved and extended a novel ac calorimetric technique for measuring the heat capacity of adsorbates on evaporated metal films. Metallic substrates are of particular interest in current studies of the thermodynamics of adsorbed molecules. The method described in the present work is only calorimetric technique which allows measurements of molecules on simple metallic surfaces. Among other improvements, we have achieved significant progress in the preparation and characterization of the evaporated metal film. We have applied this novel technique to a study of hydrogen multilayers on gold and sapphire substrates. We have shown that samples of normal-hydrogen with a nominal coverage n of approximately 25 monolayers (ML) undergo a bulk-like orientational ordering transition. The transition is suppressed as the coverage is decreased, and no sign of the transition remains above 1.6 K for n {approx} 1 ML. For n {approx_lt} 8 ML, the peak in the heat capacity exhibits signs of finite-size effects. At higher coverages, finite-size effects are not observed, and the shape of the peak depends strongly on the substrate. We conclude that the peak is inhomogeneously broadened for n {approx_lt} 8 ML. This work represents the first measurements of the heat capacity due to orientational ordering in adsorbed hydrogen. The results of an earlier experiment involving vibrational spectroscopy of adsorbed molecules are included in the Appendix. In this work, we have used infrared emission spectroscopy to study the spectral region in the vicinity of the C=O stretch vibration of bridge-bonded CO on Pt(111).

  10. AC microcalorimetry of adsorbates on evaporated metal films: Orientational ordering of H sub 2 multilayers

    SciTech Connect

    Phelps, R.B.

    1991-11-01

    We have improved and extended a novel ac calorimetric technique for measuring the heat capacity of adsorbates on evaporated metal films. Metallic substrates are of particular interest in current studies of the thermodynamics of adsorbed molecules. The method described in the present work is only calorimetric technique which allows measurements of molecules on simple metallic surfaces. Among other improvements, we have achieved significant progress in the preparation and characterization of the evaporated metal film. We have applied this novel technique to a study of hydrogen multilayers on gold and sapphire substrates. We have shown that samples of normal-hydrogen with a nominal coverage n of approximately 25 monolayers (ML) undergo a bulk-like orientational ordering transition. The transition is suppressed as the coverage is decreased, and no sign of the transition remains above 1.6 K for n {approx} 1 ML. For n {approx lt} 8 ML, the peak in the heat capacity exhibits signs of finite-size effects. At higher coverages, finite-size effects are not observed, and the shape of the peak depends strongly on the substrate. We conclude that the peak is inhomogeneously broadened for n {approx lt} 8 ML. This work represents the first measurements of the heat capacity due to orientational ordering in adsorbed hydrogen. The results of an earlier experiment involving vibrational spectroscopy of adsorbed molecules are included in the Appendix. In this work, we have used infrared emission spectroscopy to study the spectral region in the vicinity of the C=O stretch vibration of bridge-bonded CO on Pt(111).

  11. Atomic force microscopy of AgBr crystals and adsorbed gelatin films

    SciTech Connect

    Haugstad, G.; Gladfelter, W.L.; Keyes, M.P.; Weberg, E.B.

    1993-06-01

    Atomic force microscopy of the (111) surface of macroscopic AgBr crystals revealed steps ranging in height from two atomic layers up to 10 nm, lying predominantly along the (110) and (112) families of crystal directions. Rods of elemental Ag, formed via photoreduction, were observed along the (110) family of directions. Images of adsorbed gelatin films revealed circular pores with diameters of order 10-100 nm, extending to the AgBr surface. The length of deposition time, the pH and concentration of the gelatin solution, and the presence of steps on the AgBr surface were observed to affect the size, number, and location of pores in the gelatin films. 12 refs., 7 figs.

  12. Liquid 4He Adsorbed Films on Very Attractive Substrates

    NASA Astrophysics Data System (ADS)

    Urrutia, Ignacio; Szybisz, Leszek

    Adsorbed films of liquid 4He are analized, in the framework of Density Functional Theories (DF). In these systems, when the substrate becomes increasingly attractive, the thin films of 4He approaches the quasi-bidimensional limit. We study this strongly attractive substrate regime with two DF, the Orsay-Trento (OT) and a recent Hybrid proposal (Hyb), focusing in the energy behavior. It is showed that OT does not reproduce the correct limiting energy curve, and it implies that this functional could not provide reliable results for very strongly attractive substrates like Graphite (Gr). In other hand, with the Hyb DF, the correct energy behavior is found for the adsorption energy of 4He on Gr. These results show that OT should not be applied to quasi 2D (confinement) situations, and that Hyb DF provides a much more realistic description.

  13. Assembly of multilayer films incorporating a viral protein cage architecture.

    PubMed

    Suci, Peter A; Klem, Michael T; Arce, Fernando T; Douglas, Trevor; Young, Mark

    2006-10-10

    Protein cage architectures such as viral capsids, heat shock proteins, ferritins, and DNA-binding proteins are nanoscale modular subunits that can be used to expand the structural and functional range of composite materials. Here, layer-by-layer (LbL) assembly was used to incorporate cowpea chlorotic mottle virus (CCMV) into multilayer films. Three types of multilayer films were prepared. In the first type, ionic interactions were employed to assemble CCMV into triple layers. In the second type, complementary biological interactions (streptavidin/biotin) were used for this purpose. In a third variation of LbL assembly, complementary biological interactions were employed to produce nanotextured films that exhibit in-plane order over a micron scale without the need to adsorb onto a prepatterned template.

  14. Effect of peptide secondary structure on adsorption and adsorbed film properties on end-grafted polyethylene oxide layers.

    PubMed

    Binazadeh, M; Zeng, H; Unsworth, L D

    2014-01-01

    Poly-l-lysine (PLL), in α-helix or β-sheet configuration, was used as a model peptide for investigating the effect of secondary structures on adsorption events to poly(ethylene oxide) (PEO) modified surfaces formed using θ solvents. Circular dichroism results showed that the secondary structure of PLL persisted upon adsorption to Au and PEO modified Au surfaces. Quartz crystal microbalance with dissipation (QCM-D) was used to characterize the chemisorbed PEO layer in different solvents (θ and good solvents), as well as the sequential adsorption of PLL in different secondary structures (α-helix or β-sheet). QCM-D results suggest that chemisorption of PEO 750 and 2000 from θ solutions led to brushes 3.8 ± 0.1 and 4.5 ± 0.1 nm thick with layer viscosities of 9.2 ± 0.8 and 4.8 ± 0.5 cP, respectively. The average number of H2O per ethylene oxides, while in θ solvent, was determined as ~0.9 and ~1.2 for the PEO 750 and 2000 layers, respectively. Upon immersion in good solvent (as used for PLL adsorption experiments), the number of H2O per ethylene oxides increased to ~1.5 and ~2.0 for PEO 750 and 2000 films, respectively. PLL adsorbed masses for α-helix and β-sheet on Au sensors was 231 ± 5 and 1087 ± 14 ng cm(-2), with layer viscosities of 2.3 ± 0.1 and 1.2 ± 0.1 cP, respectively; suggesting that the α-helix layer was more rigid, despite a smaller adsorbed mass, than that of β-sheet layers. The PEO 750 layer reduced PLL adsorbed amounts to ~10 and 12% of that on Au for α-helices and β-sheets respectively. The PLL adsorbed mass to PEO 2000 layers dropped to ~12% and 4% of that on Au, for α-helix and β-sheet respectively. No significant differences existed for the viscosities of adsorbed α-helix and β-sheet PLL on PEO surfaces. These results provide new insights into the fundamental understanding of the effects of secondary structures of peptides and proteins on their surface adsorption.

  15. Preparation of mesoporous silica thin films by photocalcination method and their adsorption abilities for various proteins.

    PubMed

    Kato, Katsuya; Nakamura, Hitomi; Yamauchi, Yoshihiro; Nakanishi, Kazuma; Tomita, Masahiro

    2014-07-01

    Mesoporous silica (MPS) thin film biosensor platforms were established. MPS thin films were prepared from tetraethoxysilane (TEOS) via using sol-gel and spin-coating methods using a poly-(ethylene oxide)-block-poly-(propylene oxide)-block-poly-(ethylene oxide) triblock polymer, such as P123 ((EO)20(PO)70(EO)20) or F127 ((EO)106(PO)70(EO)106), as the structure-directing agent. The MPS thin film prepared using P123 as the mesoporous template and treated via vacuum ultraviolet (VUV) irradiation to remove the triblock copolymer had a more uniform pore array than that of the corresponding film prepared via thermal treatment. Protein adsorption and enzyme-linked immunosorbent assay (ELISA) on the synthesized MPS thin films were also investigated. VUV-irradiated MPS thin films adsorbed a smaller quantity of protein A than the thermally treated films; however, the human immunoglobulin G (IgG) binding efficiency was higher on the former. In addition, protein A-IgG specific binding on MPS thin films was achieved without using a blocking reagent; i.e., nonspecific adsorption was inhibited by the uniform pore arrays of the films. Furthermore, VUV-irradiated MPS thin films exhibited high sensitivity for ELISA testing, and cytochrome c adsorbed on the MPS thin films exhibited high catalytic activity and recyclability. These results suggest that MPS thin films are attractive platforms for the development of novel biosensors.

  16. Wang-Landau Simulations of Adsorbed and Confined Lattice Proteins

    NASA Astrophysics Data System (ADS)

    Pattanasiri, Busara; Li, Ying Wai; Landau, David P.; Wüst, Thomas

    2012-08-01

    The hydrophobic-polar (HP) model has emerged as one of the standard approaches for simulating protein folding. In this work, we used this model together with Wang-Landau (WL) sampling and appropriate Monte Carlo trial moves to determine the density of states and thermodynamics for two cases: Protein adsorption and protein confinement, in the vicinity of attractive surfaces. The influence on the adsorption behavior of surface attractive strength in the adsorption case and volumetric spaces in the confinement case will be discussed.

  17. The formation of standing cylinders in block copolymer films by irreversibly adsorbed polymer layers on substrates

    NASA Astrophysics Data System (ADS)

    Shang, Jun; Jiang, Naisheng; Endoh, Maya; Koga, Tadanori

    2013-03-01

    Block copolymers offer a simple and effective route to produce standing cylindrical nanostructures with regularity on the order of 10-100 nm, the length scale that is desirable for many advanced applications. However, these formations have been especially troublesome due to the fact that preferential interactions between one of the blocks and the surfaces will induce parallel alignment of the cylinders in order to minimize interfacial and surface energy. Here we introduce an alternative simple method utilizing an irreversibly adsorbed polymer layer (a ``Guiselin'' brush) as a neutral ``substrate'' formed on solid substrates for the arrangement of standing cylindrical nanostructures. The effect of polymer adsorbed layer on the long range ordering of asymmetric cylinder forming poly(styrene-block-ethylene/butylene-block-styrene) (SEBS) triblock copolymer thin films were investigated by using a combination of grazing incidence small angle x-ray scattering and atomic force microscopy techniques. We found that the SEBS, which forms cylinders lying parallel to the surface when prepared on silicon substrates, show standing cylindrical structures on selected Guiselin brush layers after prolong thermal annealing. The details will be discussed in the presentation. We acknowledges the financial support from NSF Grant No. CMMI-084626

  18. Friction and transfer of copper, silver, and gold to iron in the presence of various adsorbed surface films

    NASA Technical Reports Server (NTRS)

    Buckley, D. H.

    1979-01-01

    Sliding friction experiments were conducted with the noble metals copper, silver, and gold and two binary alloys of these metals contacting iron in the presence of various adsorbates including, oxygen, methyl mercaptan, and methyl chloride. A pin on disk specimen configuration was used with a load of 100 grams, sliding velocity of 60 mm/min; at 25 C with the surfaces saturated with the adsorbates. Auger emission spectroscopy was used to monitor surface films. Results of the experiments indicate that friction and transfer characteristics are highly specific with respect to both the noble metal and surface film present. With all three metals and films transfer of the noble metal to iron occurred very rapidly. With all metals and films transfer of the noble metal to iron continuously increased with repeated passes except for silver and copper sliding on iron sulfide.

  19. Mesoscopic Hamiltonian for the fluctuations of adsorbed Lennard-Jones liquid films.

    PubMed

    Fernández, Eva M; Chacón, Enrique; MacDowell, Luis G; Tarazona, Pedro

    2015-06-01

    We use Monte Carlo simulations of a Lennard-Jones fluid adsorbed on a short-range planar wall substrate to study the fluctuations in the thickness of the wetting layer, and we get a quantitative and consistent characterization of their mesoscopic Hamiltonian, H[ξ]. We have observed important finite-size effects, which were hampering the analysis of previous results obtained with smaller systems. The results presented here support an appealing simple functional form for H[ξ], close but not exactly equal to the theoretical nonlocal proposal made on the basis a generic density-functional analysis by Parry and coworkers. We have analyzed systems under different wetting conditions, as a proof of principle for a method that provides a quantitative bridge between the molecular interactions and the phenomenology of wetting films at mesoscopic scales.

  20. Sputtering and secondary ion emission properties of alkali metal films and adsorbed monolayers

    SciTech Connect

    Krauss, A R; Gruen, D M

    1980-01-01

    The secondary ion emission of alkali metal adsorbed monlayer and multilayer films has been studied. Profiling with sub-monolayer resolution has been performed by Auger, x-ray photoemission and secondary ion mass spectroscopy. Characteristic differences in the sputtering yields, and ion fraction have been observed which are associated with both the surface bonding properties and the mechanism leading to the formation of secondary ions. By sputtering with a negative bias applied to the sample, positive secondary ions are returned to the surface, resulting in a reduced sputter-induced erosion rate. Comparison with the results obtained with K and Li overlayers sputtered without sample bias provides an experimental value of both the total and secondary ion sputtering yields. The first and second monolayers can be readily identified and the first monolayer exhibits a lower sputtering yield and higher secondary ion fraction. This result is related to adsorption theory and measured values are compared with those obtained by thermal desorption measurements.

  1. Influence of gold nanoparticle size on the orientation and activity of adsorbed proteins

    NASA Astrophysics Data System (ADS)

    Kaur, Kanwarjeet; Forrest, James

    2010-03-01

    We used UV-visible extinction spectroscopy to study the orientation and activity of rabbit immunoglobulin G and Protein A from Staphylococcus aureus adsorbed onto gold nanoparticles of various sizes (10-60nm). There is a shift in the localised surface plasmon resonance peak due to the interaction of proteins with the nanoparticles. The proteins adopt different orientations on smaller spheres as compared to larger spheres. IgG adopts end-on orientation on bigger spheres with the Fc domain directed towards the spheres. It displays no activity towards Protein A. This study shows that the curvature of nanoparticles strongly influences the orientation of adsorbed proteins. This could be useful in the designing of colloidal drug carriers.

  2. Adsorbed Proteins Influence the Biological Activity and Molecular Targeting of Nanomaterials

    SciTech Connect

    Dutta, Debamitra; Sundaram, S. K.; Teeguarden, Justin G.; Riley, Brian J.; Fifield, Leonard S.; Jacobs, Jon M.; Addleman, Raymond S.; Kaysen, George A.; Moudgil, Brij M.; Weber, Thomas J.

    2007-11-01

    The possible combination of unique physicochemical properties operating at unique sites of action within cells and tissues has led to considerable uncertainty surrounding nanomaterial toxic potential. Here we have investigated the relative importance of proteins adsorbed onto nanomaterial surfaces in guiding uptake and toxicity to determine whether a priori identification of adsorbed proteins will contribute to nanomaterial toxicity assessment. Albumin was identified as the major protein adsorbed onto single walled carbon nanotubes (SWCNTs) following incubation with fetal bovine or human serum/plasma, but not when plasma from the Nagase Analbuminemic Rat (NAR) was used, and precoating SWCNTs with a non-ionic surfactant (Pluronic F127) inhibited albumin adsorption. Damaged or structurally altered albumin is rapidly cleared by scavenger receptors. In the RAW 264.7 macrophage-like model, we observed that SWCNTs inhibited the induction of cyclooxygenase-2 (Cox-2) by lipopolysaccharide (LPS; 1 ng/ml, 6 hr) and this anti-inflammatory response was inhibited by fucoidan (scavenger receptor antagonist) and by precoating SWCNTs with Pluronic F127. Fucoidan also reduced the uptake of fluorescent SWCNTs (Alexa647) in RAW 264.7 cells. Albumin-coated SWCNTs reduced LPS-mediated Cox-2 induction. SWCNTs did not appear to reduce binding of a fluorescent LPS (Alexa488) to RAW 264.7 cells. The profile of proteins adsorbed onto amorphous silica (50 – 1000 nm) was qualitatively different, relative to SWCNTs, and coating amorphous silica with Pluronic F127 dramatically reduced protein binding and toxicity. Collectively, these observations are consistent with an important role for adsorbed proteins in guiding nanomaterial disposition and toxicity.

  3. Structural study and wetting behavior of ethane and tetrafluoromethane thick films adsorbed on graphite (0001)

    NASA Astrophysics Data System (ADS)

    Gay, Jean-Marc; Suzanne, Jean; Pepe, Gérard; Meichel, Thierry

    1988-10-01

    We present a quantitative study of the diffraction patterns (LEED, RHEED and neutron) of ethane and tetrafluoromethane thick films adsorbed on graphite (0001). We propose to interpret the streak-like RHEED patterns of C 2H 6 and CF 4 with tabular crystallites epitaxially grown on the thin underlying film. The growth of flat ethane crystallites is explained by a partial agreement of the lattice parameters, the symmetry and the molecule orientations between the bilayer structure deduced from static energy calculations and the structure within the 3D (011) plane which appears as the interfacial plane. The change in the CF 4 RHEED pattern observed at T = 37 K and previously considered as the signature of a wetting transition might be due to a change of interfacial plane. It could be the 3D (100) or (001) plane in which a hexagonal or quasi-hexagonal symmetry in the molecule packing appears for T > 37 K. At lower temperature, T < 37 K, this symmetry could be lost with the (101¯) interfacial plane which presents a quasi-square molecule packing. We would like to emphasize the caution necessary for interpreting RHEED results. The determination of the growth mode requires the combination of different methods of measurements in order to draw conclusions without ambiguities. These two molecular systems show rather well the difficulties for interpreting experimental results on the wetting phenomenon.

  4. Effects of excluded surface area and adsorbate clustering on surface adsorption of proteins. II. Kinetic models.

    PubMed Central

    Minton, A P

    2001-01-01

    Models for equilibrium surface adsorption of proteins have been recently proposed (Minton, A. P., 2000. Biophys. Chem. 86:239-247) in which negative cooperativity due to area exclusion by adsorbate molecules is compensated to a variable extent by the formation of a heterogeneous population of monolayer surface clusters of adsorbed protein molecules. In the present work this concept is extended to treat the kinetics of protein adsorption. It is postulated that clusters may grow via two distinct kinetic pathways. The first pathway is the diffusion of adsorbed monomer to the edge of a preexisting cluster and subsequent accretion. The second pathway consists of direct deposition of a monomer in solution onto the upper (solution-facing) surface of a preexisting cluster ("piggyback" deposition) and subsequent incorporation into the cluster. Results of calculations of the time course of adsorption, carried out for two different limiting models of cluster structure and energetics, show that in the absence of piggyback deposition, enhancement of the tendency of adsorbate to cluster can reduce, but not eliminate, the negative kinetic cooperativity due to surface area exclusion by adsorbate. Apparently noncooperative (Langmuir-like) and positively cooperative adsorption progress curves, qualitatively similar to those reported in several published experimental studies, require a significant fraction of total adsorption flux through the piggyback deposition pathway. According to the model developed here and in the above-mentioned reference, the formation of surface clusters should be a common concomitant of non-site-specific surface adsorption of proteins, and may provide an important mechanism for assembly of organized "protein machines" in vivo. PMID:11259279

  5. Preparation of dye-adsorbing ZnO thin films by electroless deposition and their photoelectrochemical properties.

    PubMed

    Nagaya, Satoshi; Nishikiori, Hiromasa

    2013-09-25

    Dye-adsorbing ZnO thin films were prepared on ITO films by electroless deposition. The films were formed in an aqueous solution containing zinc nitrate, dimethylamine-borane, and eosin Y at 328 K. The film thickness was 1.2-2.0 μm. Thinner and larger-plane hexagonal columns were produced from the solution containing a higher concentration of eosin Y. A photocurrent was observed in the electrodes containing such ZnO films during light irradiation. The photoelectrochemical performance of the film was improved by increasing the concentration of eosin Y because of increases in the amount of absorbed photons and the electronic conductivity of ZnO.

  6. Controllable Nonspecific Protein Adsorption by Charged Hyperbranched Polyglycerol Thin Films.

    PubMed

    Yu, Yaming; Frey, Holger

    2015-12-08

    Antifouling thin films derived from charged hyperbranched polyglycerol (hbPG) layers were fabricated and evaluated. The anionic hbPG (a-hbPG) monolayers and cationic hbPG/anionic hbPG (c/a-hbPG) bilayers were adsorbed on the underlying self-assembled monolayers (SAMs) of cysteamine and 3-mercaptopropionic acid (3-MPA) by electrostatic interaction, respectively, and their procession was monitored by surface plasmon resonance spectroscopy (SPR). The adsorption of bovine serum albumin (BSA) and fibrinogen on the premade a-hbPG and c/a-hbPG thin films was measured and the capability of these thin films to resist nonspecific protein adsorption was evaluated by SPR as well. It is observed that the c/a-hbPG bilayer films possessed good antifouling properties. With c/a-hbPG bilayers consisting of higher molecular weight a-hbPG, the adsorption of BSA and fibrinogen were as low as 0.015 ng/mm(-2) and 0.0076 ng/mm(-2), respectively, comparable to the traditionally ultralow antifouling surfaces (<0.05 ng/mm(-2) of nonspecific protein adsorption). This work proved that the charged hbPG thin films can strongly reduce the nonspecific protein adsorption and have the promise for the antifouling coatings with improved performance.

  7. Study on Wear Reduction Mechanisms of Artificial Cartilage by Synergistic Protein Boundary Film Formation

    NASA Astrophysics Data System (ADS)

    Nakashima, Kazuhiro; Sawae, Yoshinori; Murakami, Teruo

    Poly(vinyl alcohol) (PVA) hydrogel is one of the anticipated materials for artificial cartilage. PVA hydrogel has high water content and a low elastic modulus similar to natural cartilage, but its major disadvantage is its lower strength. PVA hydrogel experienced rapid wear under severe conditions such as mixed or boundary lubrication. Therefore, the existence of a protective surface film with low friction becomes important to prevent surface failure. In this study, the reciprocating frictional tests for a sliding pair of PVA hydrogel and glass plate were carried out, and fluorescent observations were performed to identify the roles of adsorbed protein film. Albumin and γ-globulin, which are contained in natural synovial fluid, were used by mixing into the lubricant. It appears that groups of albumin molecules adsorb on the smooth γ-globulin adsorbed layer at content of 2.1wt% of proteins with an appropriate ratio. But in the case of a lubricant which has excessive protein at 2.8wt%, albumin and γ-globulin adsorbed separately. Considering the wear reduction at 2.1wt% content of protein, albumin and γ-globulin constituted synergistic adsorbed film for wear reduction. It is indicated that albumin constructs a low shear layer and γ-globulin forms a layer protecting PVA hydrogel from wear. It is considered that wear and friction of PVA hydrogel were reduced due to slip of the boundary of adsorbed albumin and γ-globulin layer. Content of protein and ratio of albumin to γ-globulin (AG ratio) are important to constitute the appropriate protein film.

  8. Tunable Adsorption and Film Formation of Mussel Adhesive Protein by Potential Control.

    PubMed

    Zhang, Fan; Xie, Guoxin; Pan, Jinshan

    2017-01-23

    Mussel adhesive proteins are of great interest in many applications because of their outstanding adhesive property and film-forming ability. Understanding and controlling the film formation and its performance is crucial for the effective use of such proteins. In this study, we focus on the potential controlled film formation and compaction of one mussel adhesive protein, Mefp-1. The adsorption and film-forming behavior of Mefp-1 on a platinum (Pt) substrate under applied potentials were investigated by cyclic voltammetry, potential-controlled electrochemical impedance spectroscopy (EIS), and quartz crystal microbalance with dissipation monitoring (QCM-D). Moreover, microfriction measurements were performed to evaluate the mechanical properties of the Mefp-1 films formed at selected potentials. The results led to the conclusion that Mefp-1 adsorbs on the Pt substrate through both electrostatic and nonelectrostatic interactions and shows an effective blocking effect for the electroactive sites on the substrate. The properties of the adsorbed Mefp-1 film vary with the applied potential, and the compactness of the adsorbed Mefp-1 film can be reversibly tuned by the applied potential.

  9. Cationized hydroxyethylcellulose as a novel, adsorbed coating for basic protein separation by capillary electrophoresis.

    PubMed

    Yang, Runmiao; Shi, Ronghua; Peng, Shuhua; Zhou, Dan; Liu, Hang; Wang, Yanmei

    2008-04-01

    We present cationized hydroxyethylcellulose (cat-HEC) synthesized in our laboratory as a novel physically adsorbed coating for CE. This capillary coating is simple and easy to obtain as it only requires flushing the capillary with polymer aqueous solution. A comparative study with and without polymers was performed. The adsorbed cat-HEC coating exhibited minimal interactions with basic proteins, providing efficient basic protein separations with excellent reproducibility. Under broad pHs, the amine groups are the main charged groups bringing about a global positive charge on the capillary wall. As a consequence, the cat-HEC coating produced an anodal EOF performance. A comparative study on the use of hydroxyethylcellulose (HEC) and cat-HEC as physically adsorbed coatings for CE are also presented. The separation efficiency and analysis reproducibility proved that the cat-HEC polymer was efficient in suppressing the adsorption of basic proteins onto the silica capillary wall. The long-term stability of the cat-HEC coating in consecutive protein separation runs has demonstrated the suitability of the coating for high-throughput electrophoretic protein separations.

  10. The effect of gold nanoparticle structure on the conformation and function of adsorbed proteins

    NASA Astrophysics Data System (ADS)

    Gagner, Jennifer E.

    Many applications of nanobiomaterials rely on or are enhanced by specific, protein-mediated interactions with biological systems. These interactions can be engineered by chemically modifying the surface of the material to affect protein adsorption, or by altering the topography of the nanoscale surface. The attachment or adsorption of proteins onto materials can greatly affect the structure and subsequent function of those proteins, giving rise to unpredictable and potentially undesirable effects. Thus, it is essential to develop a detailed understanding of how nanostructured surface characteristics, such as atomic-scale topography, surface energy, and chemical structure may affect protein adsorption, structure, function, and stability. The presented work on gold nanoparticles (AuNP) in the forms of spheres (AuNS), rods (AuNR), cubes (AuNC) and octahedra (AuNO) will elucidate the effect of nanoparticle morphology on adsorbed model proteins lysozyme (Lyz) and α-chymotrypsin (ChT). It has been found that nanoparticle morphology does affect the structure of adsorbed proteins as well as the extent of the surface coverage; however, the final form of the nano-bio conjugate is protein specific. Lyz conjugates underwent loss of structure and rapid aggregation regardless of AuNP morphology; however, ChT conjugates exhibited no structure loss when immobilized on AuNS, and a significant, loading specific structure loss when adsorbed on AuNR. Further work will be presented on efforts to determine the role of crystal structure, surface energy, and ligand chemistry on adsorbed proteins. Wet chemical methods are used to synthesize AuNC with f100g facets and AuNO with f111g facets. Nanoparticles are characterized through electron microscopy, X-ray and electron diffraction, X-ray photoelectron spectroscopy and inductively coupled plasma mass spectroscopy. Protein conjugation and changes in protein structure are monitored through a variety of physical and spectroscopic techniques

  11. Single and binary adsorption of proteins on ion-exchange adsorbent: The effectiveness of isothermal models.

    PubMed

    Liang, Juan; Fieg, Georg; Shi, Qing-Hong; Sun, Yan

    2012-09-01

    Simultaneous and sequential adsorption equilibria of single and binary adsorption of bovine serum albumin and bovine hemoglobin on Q Sepharose FF were investigated in different buffer constituents and initial conditions. The results in simultaneous adsorption showed that both proteins underwent competitive adsorption onto the adsorbent following greatly by protein-surface interaction. Preferentially adsorbed albumin complied with the universal rule of ion-exchange adsorption whereas buffer had no marked influence on hemoglobin adsorption. Moreover, an increase in initial ratios of proteins was benefit to a growth of adsorption density. In sequential adsorption, hemoglobin had the same adsorption densities as single-component adsorption. It was attributed to the displacement of preadsorbed albumin and multiple layer adsorption of hemoglobin. Three isothermal models (i.e. extended Langmuir, steric mass-action, and statistical thermodynamic (ST) models) were introduced to describe the ion-exchange adsorption of albumin and hemoglobin mixtures. The results suggested that extended Langmuir model gave the lowest deviation in describing preferential adsorption of albumin at a given salt concentration while steric mass-action model could very well describe the salt effect in albumin adsorption. For weaker adsorbed hemoglobin, ST model was the preferred choice. In concert with breakthrough data, the research further revealed the complexity in ion-exchange adsorption of proteins.

  12. Melt crystallization/dewetting of ultrathin PEO films via carbon dioxide annealing: the effects of polymer adsorbed layers.

    PubMed

    Asada, Mitsunori; Jiang, Naisheng; Sendogdular, Levent; Sokolov, Jonathan; Endoh, Maya K; Koga, Tadanori; Fukuto, Masafumi; Yang, Lin; Akgun, Bulent; Dimitriou, Michael; Satija, Sushil

    2014-09-14

    The effects of CO2 annealing on the melting and subsequent melt crystallization processes of spin-cast poly(ethylene oxide) (PEO) ultrathin films (20-100 nm in thickness) prepared on Si substrates were investigated. By using in situ neutron reflectivity, we found that all the PEO thin films show melting at a pressure as low as P = 2.9 MPa and at T = 48 °C which is below the bulk melting temperature (Tm). The films were then subjected to quick depressurization to atmospheric pressure, resulting in the non-equilibrium swollen state, and the melt crystallization (and/or dewetting) process was carried out in air via subsequent annealing at given temperatures below Tm. Detailed structural characterization using grazing incidence X-ray diffraction, atomic force microscopy, and polarized optical microscopy revealed two unique aspects of the CO2-treated PEO films: (i) a flat-on lamellar orientation, where the molecular chains stand normal to the film surface, is formed within the entire film regardless of the original film thickness and the annealing temperature; and (ii) the dewetting kinetics for the 20 nm thick film is much slower than that for the thicker films. The key to these phenomena is the formation of irreversibly adsorbed layers on the substrates during the CO2 annealing: the limited plasticization effect of CO2 at the polymer-substrate interface promotes polymer adsorption rather than melting. Here we explain the mechanisms of the melt crystallization and dewetting processes where the adsorbed layers play vital roles.

  13. [Interactions between proteins and cation exchange adsorbents analyzed by NMR and hydrogen/deuterium exchange technique].

    PubMed

    Wang, Kang; Hao, Dongxia; Qi, Shuting; Ma, Guanghui

    2014-09-01

    In silico acquirement of the accurate residue details of protein on chromatographic media is a bottleneck in protein chromatography separation and purification. Here we developed a novel approach by coupling with H/D exchange and nuclear magnetic resonance to observe hen egg white lysozyme (HEWL) unfolding behavior adsorbed on cation exchange media (SP Sepharose FF). Analysis of 1D 1H-NMR shows that protein unfolding accelerated H/D exchange rate, leading to more loss of signal of amide hydrogen owing to exposure of residues and the more unfolding of protein. Analysis of two-dimensional hydrogen-hydrogen total correlation spectroscopy shows that lysozyme lost more signals and experienced great unfolding during its adsorption on media surface. However, for several distinct fragments, the protection degrees varied, the adsorbed lysozyme lost more signal intensity and was less protected at disorder structures (coil, bend, and turn), but was comparatively more protected against exchange at secondary structure domains (α-helix, β-sheet). Finally, the binding site was determined by electrostatic calculations using computer simulation methods in conjunction with hydrogen deuterium labeled protein and NMR. This study would help deeply understand the microscopic mechanism of protein chromatography and guide the purposely design of chromatographic process and media. Moreover, it also provide an effective tool to study the protein and biomaterials interaction in other applications.

  14. A simplified density functional theory method for investigating charged adsorbates on an ultrathin, insulating film supported by a metal substrate.

    PubMed

    Scivetti, Ivan; Persson, Mats

    2014-04-02

    A simplified density functional theory (DFT) method for investigating charged adsorbates on an ultrathin, insulating film supported by a metal substrate is developed and presented. This new method is based on a previous DFT development that uses a perfect conductor (PC) model to approximate the electrostatic response of the metal substrate, while the film and the adsorbate are both treated fully within DFT (Scivetti and Persson 2013 J. Phys.: Condens. Matter 25 355006). The missing interactions between the metal substrate and the insulating film in the PC approximation are modelled by a simple force field (FF). The parameters of the PC model and the force field are obtained from DFT calculations of the film and the substrate, here shown explicitly for a NaCl bilayer supported by a Cu(100) surface. In order to obtain some of these parameters and the polarizability of the force field, we have to include an external, uniformly charged plane in the DFT calculations, which has required the development of a periodic DFT formalism to include such a charged plane in the presence of a metal substrate. This extension and implementation should be of more general interest and applicable to other challenging problems, for instance, in electrochemistry. As illustrated for the gold atom on the NaCl bilayer supported by a Cu(100) surface, our new DFT-PC-FF method allows us to handle different charge states of adsorbates in a controlled and accurate manner with a considerable reduction of the computational time. In addition, it is now possible to calculate vertical transition and reorganization energies for the charging and discharging of adsorbates that cannot be obtained by current DFT methodologies that include the metal substrate. We find that the computed vertical transition energy for charging of the gold adatom is in good agreement with experiments.

  15. Aging of the nanosized photochromic WO3 films and the role of adsorbed water in the photochromism

    NASA Astrophysics Data System (ADS)

    Gavrilyuk, A. I.

    2016-02-01

    Here it has been reported on aging of the nanosized WO3 film, which is revealed is continuous reduction of the photochromic sensitivity over time. Water molecules physically adsorbed on the film surface from ambient air form donor-acceptor and hydrogen bonds, changing gradually the adsorption state to chemisorption which prevents an access of organic molecules that serve as hydrogen donors by the photochromism. The mechanism of the process has been investigated and discussed. The role of water in the photochromism has been highlighted. The difference in the efficiency for being of a hydrogen donor in the photochromic process between water and organic molecules is discussed.

  16. Modeling colloid deposition on a protein layer adsorbed to iron-oxide-coated sand

    NASA Astrophysics Data System (ADS)

    Yang, X.; Flynn, R.; von der Kammer, F.; Hofmann, T.

    2012-11-01

    Our recent study reported that conformation change of granule-associated Bovine Serum Albumin (BSA) may influence the role of the protein controlling colloid deposition in porous media (Flynn et al., 2012). The present study conceptualized the observed phenomena with an ellipsoid morphology model, describing BSA as an ellipsoid taking a side-on or end-on conformation on granular surface, and identified the following processes: (1) at low adsorbed concentrations, BSA exhibited a side-on conformation blocking colloid deposition; (2) at high adsorbed concentrations, BSA adapted to an end-on conformation promoted colloid deposition; and (3) colloid deposition on the BSA layer may progressively generate end-on molecules (sites) by conformation change of side-on BSA, resulting in sustained increasing deposition rates. Generally, the protein layer lowered colloid attenuation by the porous medium, suggesting the overall effect of BSA was inhibitory at the experimental time scale. A mathematical model was developed to interpret the ripening curves. Modeling analysis identified the site generation efficiency of colloid as a control on the ripening rate (declining rate in colloid concentrations), and this efficiency was higher for BSA adsorbed from a more dilute BSA solution.

  17. Differential analysis of "protein corona" profile adsorbed onto different nonviral gene delivery systems.

    PubMed

    Capriotti, Anna Laura; Caracciolo, Giulio; Caruso, Giuseppe; Foglia, Patrizia; Pozzi, Daniela; Samperi, Roberto; Laganà, Aldo

    2011-12-15

    A shotgun proteomics approach was used to characterize and compare the proteins that lead to the formation of a rich "protein corona" adsorbed onto the surfaces of cationic liposomes (CLs), lipoplexes, and lipid/polycation/DNA (LPD) complexes, when they come into contact with plasma. After separation of the nanoparticle-protein complex from plasma, the protein mixture was digested, and peptides were analyzed by nanoliquid chromatography-Orbitrap LTQ-XL mass spectrometry. The number of proteins bound to lipoplexes was double that of those identified in the corona of CLs (208 vs 105), while 77 proteins were common to both coronas. The number of proteins bound to the surface of the LPD complexes (158, 133 of which are common to lipoplexes) is intermediate between those found in the protein corona of both CLs and lipoplexes. About half of them were found in the protein corona of CLs. By overlapping the three formulations, it can be seen that only 12 proteins are peculiar to LPD complexes. These results may help in designing gene delivery systems capable of binding the minimum possible quantity of proteins that influence transfection negatively, binding selectively proteins capable of helping in steering in vivo the vector toward the target, and obtaining more efficient and effective gene therapy.

  18. Determinants of Protein Elution Rates from Preparative Ion-Exchange Adsorbents

    PubMed Central

    Angelo, James M.; Lenhoff, Abraham M.

    2016-01-01

    The rate processes involved in elution in preparative chromatography can affect both peak resolution and hence selectivity as well as practical factors such as facility fit. These processes depend on the physical structure of the adsorbent particles, the amount of bound solute, the solution conditions for operation or some combination of these factors. Ion-exchange adsorbents modified with covalently attached or grafted polymer layers have become widely used in preparative chromatography. Their often easily accessible microstructures offer substantial binding capacities for biomolecules, but elution has sometimes been observed to be undesirably slow. In order to determine which physicochemical phenomena control elution behavior, commercially available cellulosic, dextran-grafted and unmodified agarose materials were characterized here by their uptake and elution profiles at various conditions, including different degrees of loading. Elution data were analyzed under the assumption of purely diffusion-limited control, including the role of pore structure properties such as porosity and tortuosity. In general, effective elution rates decreased with the reduction of accessible pore volume, but differences among different proteins indicated the roles of additional factors. Additional measurements and analysis, including the use of confocal laser scanning microscopy to observe elution within single chromatographic particles, indicated the importance of protein association within the particle during elution. The use of protein stabilizing agents was explored in systems presenting atypical elution behavior, and L-arginine and disaccharide excipients were shown to alleviate the effects for one protein, lysozyme, in the presence of sodium chloride. Incorporation of these excipients into eluent buffer gave rise to faster elution and significantly lower pool volumes in elution from polymer-modified adsorbents. PMID:26948763

  19. Determinants of protein elution rates from preparative ion-exchange adsorbents.

    PubMed

    Angelo, James M; Lenhoff, Abraham M

    2016-04-01

    The rate processes involved in elution in preparative chromatography can affect both peak resolution and hence selectivity as well as practical factors such as facility fit. These processes depend on the physical structure of the adsorbent particles, the amount of bound solute, the solution conditions for operation or some combination of these factors. Ion-exchange adsorbents modified with covalently attached or grafted polymer layers have become widely used in preparative chromatography. Their often easily accessible microstructures offer substantial binding capacities for biomolecules, but elution has sometimes been observed to be undesirably slow. In order to determine which physicochemical phenomena control elution behavior, commercially available cellulosic, dextran-grafted and unmodified agarose materials were characterized here by their elution profiles at various conditions, including different degrees of loading. Elution data were analyzed under the assumption of purely diffusion-limited control, including the role of pore structure properties such as porosity and tortuosity. In general, effective elution rates decreased with the reduction of accessible pore volume, but differences among different proteins indicated the roles of additional factors. Additional measurements and analysis, including the use of confocal laser scanning microscopy to observe elution within single chromatographic particles, indicated the importance of protein association within the particle during elution. The use of protein stabilizing agents was explored in systems presenting atypical elution behavior, and l-arginine and disaccharide excipients were shown to alleviate the effects for one protein, lysozyme, in the presence of sodium chloride. Incorporation of these excipients into eluent buffer gave rise to faster elution and significantly lower pool volumes in elution from polymer-modified adsorbents.

  20. Physical and chemical characterization of adsorbed protein onto gold electrode functionalized with Tunisian coral and nacre.

    PubMed

    Hamza, Samir; Bouchemi, Meryem; Slimane, Noureddine; Azari, Zitouni

    2013-01-01

    Bone substitutes are more and more used in bone surgery because of their biologic safety, clinic efficiency and facility to synthesize. Bone substitutes with active osteogenic properties, associating biomaterials with organic macromolecule components of the extracellular matrix (protein, GAG) are recommended. Nevertheless, we should have a simple technique to control interactions between proteins and the material. Natural coral and nacre have been found to be impressive bone graft substitutes. In this work, we characterize nacre and coral powder using energy dispersive X-ray analysis (EDX). We used electrochemical impedance spectroscopy (EIS) and attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy to evaluate bovine serum albumin (BSA) as model protein, adsorbed to these biomaterial surfaces. In order to understand the nacre/coral-protein interfacial compatibility, it is necessary to investigate the wettability.

  1. Isolation of calcium-binding proteins on selective adsorbents. Application to purification of bovine calmodulin.

    PubMed

    Chaga, G S; Ersson, B; Porath, J O

    1996-05-03

    We report the fractionation of calcium-binding proteins using immobilized metal ion affinity chromatography (IMAC) with hard metal ions. Various hard metal ions (Mn2+, La3+, Nd3+, Eu(3 were immobilized on cross-linked agarose substituted with Tris(carboxymethyl)ethylenediamine (TED) and used as an adsorbent. After systematic studies, europium was selected for further work on the fractionation of calcium-binding proteins. It was found that the presence of Ca2+ in the sample and the solvent strongly promoted the adsorption and selectivity. Selective elution was accomplished in stepwise mode by the addition of calcium chelators such as malonate, citrate and phosphate. Calmodulin of high purity was isolated from a crude extract. Similar behavior of other calcium-binding proteins indicates that the reported chromatographic procedure can be generally applied to such proteins.

  2. The leucine rich amelogenin protein (LRAP) adsorbs as monomers or dimers onto surfaces

    SciTech Connect

    Tarasevich, Barbara J.; Lea, Alan S.; Shaw, Wendy J.

    2010-03-15

    Amelogenin and amelogenin splice variants are believed to be involved in controlling the formation of the highly anisotropic and ordered hydroxyapatite crystallites that form enamel. The adsorption behavior of amelogenin proteins onto substrates is very important because protein-surface interactions are critical to it’s function. We have studied the adsorption of LRAP, a splice variant of amelogenin which may also contribute to enamel function, onto model self-assembled monolayers on gold containing of COOH, CH3, and NH2 end groups. Dynamic light scattering (DLS) experiments indicated that LRAP in phosphate buffered saline (PBS) and solutions at saturation with calcium phosphate contained aggregates of nanospheres. Null ellipsometry and atomic force microscopy (AFM) were used to study protein adsorption amounts and structures. Relatively high amounts of adsorption occurred onto the CH3 and NH2 surfaces from both calcium phosphate and PBS solutions. Adsorption was also promoted onto COOH surfaces when calcium was present in the solutions suggesting an interaction that involves calcium bridging with the negatively charged C-terminus. The ellipsometry and AFM studies suggested that the protein adsorbed onto all surfaces as LRAP monomers. We propose that the monomers adsorb onto the surfaces by disassembling or “shedding” from the nanospheres that are present in solution. This work reveals the importance of small subnanosphere-sized structures of LRAP at interfaces, structures that may be important in the biomineralization of tooth enamel.

  3. Protein-Adsorbed Magnetic-Nanoparticle-Mediated Assay for Rapid Detection of Bacterial Antibiotic Resistance.

    PubMed

    Cowger, Taku A; Yang, Yaping; Rink, David E; Todd, Trever; Chen, Hongmin; Shen, Ye; Yan, Yajun; Xie, Jin

    2017-02-17

    Antibiotic susceptibility tests have been used for years as a crucial diagnostic tool against antibiotic-resistant bacteria. However, due to a lack of biomarkers specific to resistant types, these approaches are often time-consuming, inaccurate, and inflexible in drug selections. Here, we present a novel susceptibility test method named protein-adsorbed nanoparticle-mediated matrix-assisted laser desorption-ionization mass spectrometry, or PANMS. Briefly, we adsorb five different proteins (β-casein, α-lactalbumin, human serum albumin, fibrinogen, and avidin) onto the surface of Fe3O4. Upon interaction with bacteria surface, proteins were displaced from the nanoparticle surface, the amounts of which were quantified by matrix-assisted laser desorption ionization mass spectrometry. We find that the protein displacement profile was different distinctive among different bacteria strains and, in particular, between wild-type and drug-resistant strains. More excitingly, we observe bacteria resistant to drugs of the same mechanisms share similar displacement profiles on a linear discriminant analysis (LDA) map. This suggests the possibility of using PANMS to identify the type of mechanism behind antibiotic resistance, which was confirmed in a blind test. Given that PANMS is free of drug incubation and the whole procedure takes less than 50 min, it holds great potential as a high-throughput, low-cost, and accurate drug susceptibility test in the clinic.

  4. Development of a rapid sanitization solution for silica-based protein A affinity adsorbents.

    PubMed

    Rogers, Marc; Hiraoka-Sutow, Martha; Mak, Polly; Mann, Fred; Lebreton, Bénédicte

    2009-05-22

    Protein A chromatography media require sanitization between batches as well as prior to long-term storage. While sodium hydroxide (NaOH) is probably one of the most widely used sanitants within the bioprocess industry, it cannot be used with silica- or controlled pore glass (CPG)-based adsorbents due to the instability of the base matrix at high pH. Benzyl alcohol is commonly used for sanitizing such adsorbents, though extended contact times may be required to meet desired microbial log reduction values, especially for fungal and bacterial spore formers. With the rising market need for monoclonal antibody therapeutics, higher manufacturing throughput may be required. In such cases, a shorter sanitization cycle would be extremely beneficial to maximize manufacturing throughput and productivity. This paper describes the development of a new synergistic sanitant solution, designated PAB (120 mM phosphoric acid, 167 mM acetic acid, 2.2% benzyl alcohol) that delivers improved microbial kill kinetics, enabling sanitization times of 2-3h at room temperature, while maintaining acceptable adsorbent stability. Both the approaches taken to establish the effectiveness of the improved solution as well as confirmation of its process compatibility are covered here.

  5. Application of immobilized metal ion chelate complexes as pseudocation exchange adsorbents for protein separation.

    PubMed

    Zachariou, M; Hearn, M T

    1996-01-09

    The interactions of horse muscle myoglobin (MYO), tuna heart cytochrome c (CYT), and hen egg white lysozyme (LYS) with three different immobilized metal ion affinity (IMAC) adsorbents involving the chelated complexes of the hard Lewis metal ions Al3+, Ca2+, Fe3+, and Yb3+ and the borderline Lewis metal ion Cu2+ have been investigated in the presence of low- and high-ionic strength buffers and at two different pH values. In contrast to the selectivity behavior noted with buffers of high ionic strength, with low-ionic strength buffers, these three proteins interact with the hard metal ion IMAC adsorbents in a manner more characteristic of cation exchange behavior, although in contrast to the cation exchange chromatography of these proteins, as the pH value of the elution buffer was increased, the retention also increased. The selectivity differences observed under these conditions appear to be due to the formation of hydrolytic complexes of these immobilized metal ion chelate systems involving a change in the coordination geometry of the im-M(n+)-chelate at higher pH values. The experimental observations have been evaluated in terms of the effective charge on the immobilized metal ion chelate complex and the charge characteristics of the specific proteins.

  6. Shotgun proteomic analytical approach for studying proteins adsorbed onto liposome surface.

    PubMed

    Capriotti, Anna Laura; Caracciolo, Giulio; Cavaliere, Chiara; Crescenzi, Carlo; Pozzi, Daniela; Laganà, Aldo

    2011-09-01

    The knowledge about the interaction between plasma proteins and nanocarriers employed for in vivo delivery is fundamental to understand their biodistribution. Protein adsorption onto nanoparticle surface (protein corona) is strongly affected by vector surface characteristics. In general, the primary interaction is thought to be electrostatic, thus surface charge of carrier is supposed to play a central role in protein adsorption. Because protein corona composition can be critical in modifying the interactive surface that is recognized by cells, characterizing its formation onto lipid particles may serve as a fundamental predictive model for the in vivo efficiency of a lipidic vector. In the present work, protein coronas adsorbed onto three differently charged cationic liposome formulations were compared by a shotgun proteomic approach based on nano-liquid chromatography-high-resolution mass spectrometry. About 130 proteins were identified in each corona, with only small differences between the different cationic liposome formulations. However, this study could be useful for the future controlled design of colloidal drug carriers and possibly in the controlled creation of biocompatible surfaces of other devices that come into contact with proteins into body fluids.

  7. Conformational transition free energy profiles of an adsorbed, lattice model protein by multicanonical Monte Carlo simulation

    NASA Astrophysics Data System (ADS)

    Castells, Victoria; Van Tassel, Paul R.

    2005-02-01

    Proteins often undergo changes in internal conformation upon interacting with a surface. We investigate the thermodynamics of surface induced conformational change in a lattice model protein using a multicanonical Monte Carlo method. The protein is a linear heteropolymer of 27 segments (of types A and B) confined to a cubic lattice. The segmental order and nearest neighbor contact energies are chosen to yield, in the absence of an adsorbing surface, a unique 3×3×3 folded structure. The surface is a plane of sites interacting either equally with A and B segments (equal affinity surface) or more strongly with the A segments (A affinity surface). We use a multicanonical Monte Carlo algorithm, with configuration bias and jump walking moves, featuring an iteratively updated sampling function that converges to the reciprocal of the density of states 1/Ω(E), E being the potential energy. We find inflection points in the configurational entropy, S(E)=klnΩ(E), for all but a strongly adsorbing equal affinity surface, indicating the presence of free energy barriers to transition. When protein-surface interactions are weak, the free energy profiles F(E)=E-TS(E) qualitatively resemble those of a protein in the absence of a surface: a free energy barrier separates a folded, lowest energy state from globular, higher energy states. The surface acts in this case to stabilize the globular states relative to the folded state. When the protein surface interactions are stronger, the situation differs markedly: the folded state no longer occurs at the lowest energy and free energy barriers may be absent altogether.

  8. Effects of surface curvature and surface chemistry on the structure and activity of proteins adsorbed in nanopores.

    PubMed

    Sang, Lung-Ching; Coppens, Marc-Olivier

    2011-04-14

    The interactions of proteins with the surface of cylindrical nanopores are systematically investigated to elucidate how surface curvature and surface chemistry affect the conformation and activity of confined proteins in an aqueous, buffered environment. Two globular proteins, lysozyme and myoglobin, with different catalytic functions, were used as model proteins to analyze structural changes in proteins after adsorption on ordered mesoporous silica SBA-15 and propyl-functionalized SBA-15 (C(3)SBA-15) with carefully controlled pore size. Liquid phase ATR-FTIR spectroscopy was used to study the amide I and II bands of the adsorbed proteins. The amide I bands showed that the secondary structures of free and adsorbed protein molecules differ, and that the secondary structure of the adsorbed protein is influenced by the local geometry as well as by the surface chemistry of the nanopores. The conformation of the adsorbed proteins inside the nanopores of SBA-15 and C(3)SBA-15 is strongly correlated with the local geometry and the surface properties of the nanoporous materials, which results in different catalytic activities. Adsorption by electrostatic interaction of proteins in nanopores of an optimal size provides a favorably confining and protecting environment, which may lead to considerably enhanced structural stability and catalytic activity.

  9. ToF-SIMS Analysis of Adsorbed Proteins: Principal Component Analysis of the Primary Ion Species Effect on the Protein Fragmentation Patterns.

    PubMed

    Muramoto, Shin; Graham, Daniel J; Wagner, Matthew S; Lee, Tae Geol; Moon, Dae Won; Castner, David G

    2011-12-15

    In time-of-flight secondary ion mass spectrometry (ToF-SIMS), the choice of primary ion used for analysis can influence the resulting mass spectrum. This is because different primary ion types can produce different fragmentation pathways. In this study, analysis of single-component protein monolayers were performed using monatomic, tri-atomic, and polyatomic primary ion sources. Eight primary ions (Cs(+), Au(+), Au(3) (+), Bi(+), Bi(3) (+), Bi(3) (++), C(60) (+)) were used to examine to the low mass (m/z < 200) fragmentation patterns from five different proteins (bovine serum albumin, bovine serum fibrinogen, bovine immunoglobulin G and chicken egg white lysozyme) adsorbed onto mica surfaces. Principal component analysis (PCA) processing of the ToF-SIMS data showed that variation in peak intensity caused by the primary ions was greater than differences in protein composition. The spectra generated by Cs(+), Au(+) and Bi(+) primary ions were similar, but the spectra generated by monatomic, tri-atomic and polyatomic primary ion ions varied significantly. C(60) primary ions increased fragmentation of the adsorbed proteins in the m/z < 200 region, resulting in more intense low m/z peaks. Thus, comparison of data obtained by one primary ion species with that obtained by another primary ion species should be done with caution. However, for the spectra generated using a given primary ion beam, discrimination between the spectra of different proteins followed similar trends. Therefore, a PCA model of proteins created with a given ion source should only be applied to datasets obtained using the same ion source. The type of information obtained from PCA depended on the peak set used. When only amino acid peaks were used, PCA was able to identify the relationship between proteins by their amino acid composition. When all peaks from m/z 12-200 were used, PCA separated proteins based on a ratio of C(4)H(8)N(+) to K(+) peak intensities. This ratio correlated with the thickness

  10. Measurement of interactions between protein layers adsorbed on silica by atomic force microscopy

    NASA Astrophysics Data System (ADS)

    Valle-Delgado, J. J.; Molina-Bolívar, J. A.; Galisteo-González, F.; Gálvez-Ruiz, M. J.; Feiler, A.; Rutland, M. W.

    2004-07-01

    The present work, using an atomic force microscope and the colloid probe technique, investigates the interaction forces between bovine serum albumin (BSA) layers and between apoferritin layers adsorbed on silica surfaces. The measurements have been carried out in an aqueous medium at different pH values and NaCl concentrations. Similar behaviours have been found with both proteins. Electrostatic and steric forces dominate the interactions between the protein layers at low NaCl concentrations. However, a very strange behaviour is found as a function of pH at high NaCl concentrations. The results obtained under these conditions could be explained if the presence of hydration forces in these systems is assumed.

  11. The profile of adsorbed plasma and serum proteins on methacrylic acid copolymer beads: Effect on complement activation.

    PubMed

    Wells, Laura A; Guo, Hongbo; Emili, Andrew; Sefton, Michael V

    2017-02-01

    Polymer beads made of 45% methacrylic acid co methyl methacrylate (MAA beads) promote vascular regenerative responses in contrast to control materials without methacrylic acid (here polymethyl methacrylate beads, PMMA). In vitro and in vivo studies suggest that MAA copolymers induce differences in macrophage phenotype and polarization and inflammatory responses, presumably due to protein adsorption differences between the beads. To explore differences in protein adsorption in an unbiased manner, we used high resolution shotgun mass spectrometry to identify and compare proteins that adsorb from human plasma or serum onto MAA and PMMA beads. From plasma, MAA beads adsorbed many complement proteins, such as C1q, C4-related proteins and the complement inhibitor factor H, while PMMA adsorbed proteins, such as albumin, C3 and apolipoproteins. Because of the differences in complement protein adsorption, follow-up studies focused on using ELISA to assess complement activation. When incubated in serum, MAA beads generated significantly lower levels of soluble C5b9 and C3a/C3adesarg in comparison to PMMA beads, indicating a decrease in complement activation with MAA beads. The differences in adsorbed protein on the two materials likely alter subsequent cell-material interactions that ultimately result in different host responses and local vascularization.

  12. Targeted Mutagenesis and Combinatorial Library Screening Enables Control of Protein Orientation on Surfaces and Increased Activity of Adsorbed Proteins.

    PubMed

    Cruz-Teran, Carlos A; Carlin, Kevin B; Efimenko, Kirill; Genzer, Jan; Rao, Balaji M

    2016-08-30

    While nonspecific adsorption is widely used for immobilizing proteins on solid surfaces, the random nature of protein adsorption may reduce the activity of immobilized proteins due to occlusion of the active site. We hypothesized that the orientation a protein assumes on a given surface can be controlled by systematically introducing mutations into a region distant from its active site, thereby retaining activity of the immobilized protein. To test this hypothesis, we generated a combinatorial protein library by randomizing six targeted residues in a binding protein derived from highly stable, nonimmunoglobulin Sso7d scaffold; mutations were targeted in a region that is distant from the binding site. This library was screened to isolate binders that retain binding to its cognate target (chicken immunoglobulin Y, cIgY) as well as exhibit adsorption on unmodified silica at pH 7.4 and high ionic strength conditions. A single mutant, Sso7d-2B5, was selected for further characterization. Sso7d-2B5 retained binding to cIgY with an apparent dissociation constant similar to that of the parent protein; both mutant and parent proteins saturated the surface of silica with similar densities. Strikingly, however, silica beads coated with Sso7d-2B5 could achieve up to 7-fold higher capture of cIgY than beads coated with the parent protein. These results strongly suggest that mutations introduced in Sso7d-2B5 alter its orientation relative to the parent protein, when adsorbed on silica surfaces. Our approach also provides a generalizable strategy for introducing mutations in proteins so as to improve their activity upon immobilization, and has direct relevance to development of protein-based biosensors and biocatalysts.

  13. Optical and electrical characterizations of nanocomposite film of titania adsorbed onto oxidized multiwalled carbon nanotubes

    NASA Astrophysics Data System (ADS)

    Feng, Wei; Feng, Yiyu; Wu, Zigang; Fujii, Akihiko; Ozaki, Masanori; Yoshino, Katsumi

    2005-07-01

    Composite film containing titania electrostatically linked to oxidized multiwalled carbon nanotubes (TiO2-s-MWNTs) was prepared from a suspension of TiO2 nanoparticles in soluble carbon nanotubes. The structure of the film was analysed principally by Fourier transform infrared spectroscopy, scanning electron micrography and x-ray diffraction. The optical and electrical characterizations of the film were investigated by UV-vis spectrum, photoluminescence and photoconductivity. The enhancement of photocurrent in the TiO2-s-MWNT film is discussed by taking the photoinduced charge transfer between the MWNT and TiO2 into consideration.

  14. Dependence of cellular activity at protein adsorbed biointerfaces with nano- to microscale dimensionality.

    PubMed

    Nune, C; Misra, R D K; Somani, M C; Karjalainen, L P

    2014-06-01

    Protein adsorption is one of the first-few events that occur when a biomedical device comes in contact with the physiological system. The adsorption process is subsequently followed by communication with cells and formation of tissue. Given the strong interest in nanostructured surfaces, we describe here the impact of grain structure from nanograined (NG) regime to coarse-grained (CG) regime on the self-assembly of proteins (bovine serum albumin) and consequent functional response of osteoblasts. The objective is accomplished using the innovative concept of "phase reversion" that enables a wide range of grain size (from NG to CG regime) to be obtained using an identical set of parameters, besides additional attributes of high strength/weight ratio and wear resistance. Depending on the grain structure a consistent and significant change in the adsorption characteristics of protein was observed at biointerface, such that the cell density was statistically different. The high surface coverage and leaf-like conformation of adsorbed protein on NG surface as compared to bare branch-like structure with low surface coverage on the CG surface, was beneficial in favorably modulating cellular activity (osteoblast functions: cell attachment, proliferation, actin, vinculin, and fibronectin expression). This is the first report that elucidates the impact of grain structure from NG to CG regime on cellular activity.

  15. Exploring the interfacial structure of protein adsorbates and the kinetics of protein adsorption: an in situ high-energy X-ray reflectivity study.

    PubMed

    Evers, Florian; Shokuie, Kaveh; Paulus, Michael; Sternemann, Christian; Czeslik, Claus; Tolan, Metin

    2008-09-16

    The high energy X-ray reflectivity technique has been applied to study the interfacial structure of protein adsorbates and protein adsorption kinetics in situ. For this purpose, the adsorption of lysozyme at the hydrophilic silica-water interface has been chosen as a model system. The structure of adsorbed lysozyme layers was probed for various aqueous solution conditions. The effect of solution pH and lysozyme concentration on the interfacial structure was measured. Monolayer formation was observed for all cases except for the highest concentration. The adsorbed protein layers consist of adsorbed lysozyme molecules with side-on or end-on orientation. By means of time-dependent X-ray reflectivity scans, the time-evolution of adsorbed proteins was monitored as well. The results of this study demonstrate the capabilities of in situ X-ray reflectivity experiments on protein adsorbates. The great advantages of this method are the broad wave vector range available and the high time resolution.

  16. Controlled surface chemistry of diamond/β-SiC composite films for preferential protein adsorption.

    PubMed

    Wang, Tao; Handschuh-Wang, Stephan; Yang, Yang; Zhuang, Hao; Schlemper, Christoph; Wesner, Daniel; Schönherr, Holger; Zhang, Wenjun; Jiang, Xin

    2014-02-04

    Diamond and SiC both process extraordinary biocompatible, electronic, and chemical properties. A combination of diamond and SiC may lead to highly stable materials, e.g., for implants or biosensors with excellent sensing properties. Here we report on the controllable surface chemistry of diamond/β-SiC composite films and its effect on protein adsorption. For systematic and high-throughput investigations, novel diamond/β-SiC composite films with gradient composition have been synthesized using the hot filament chemical vapor deposition (HFCVD) technique. As revealed by scanning electron microscopy (SEM), the diamond/β-SiC ratio of the composite films shows a continuous change from pure diamond to β-SiC over a length of ∼ 10 mm on the surface. X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS) was employed to unveil the surface termination of chemically oxidized and hydrogen treated surfaces. The surface chemistry of the composite films was found to depend on diamond/β-SiC ratio and the surface treatment. As observed by confocal fluorescence microscopy, albumin and fibrinogen were preferentially adsorbed from buffer: after surface oxidation, the proteins preferred to adsorb on diamond rather than on β-SiC, resulting in an increasing amount of proteins adsorbed to the gradient surfaces with increasing diamond/β-SiC ratio. By contrast, for hydrogen-treated surfaces, the proteins preferentially adsorbed on β-SiC, leading to a decreasing amount of albumin adsorbed on the gradient surfaces with increasing diamond/β-SiC ratio. The mechanism of preferential protein adsorption is discussed by considering the hydrogen bonding of the water self-association network to OH-terminated surfaces and the change of the polar surface energy component, which was determined according to the van Oss method. These results suggest that the diamond/β-SiC gradient film can be a promising material for biomedical applications which

  17. Characterization of crosslinked artificial protein films

    NASA Astrophysics Data System (ADS)

    Nowatzki, Paul

    Genetically engineered artificial proteins are promising candidates for new biomaterials because their amino acid sequences can be precisely controlled. This work describes the characterization of crosslinked films of biomimetic artificial extracellular matrix (aECM) proteins with hybrid functions designed to meet materials needs in applications such as small diameter vascular grafts and corneal tissue implants. Elastin-derived polypeptides give the proteins flexibility, while RGD and CS5 peptide domains from fibronectin serve to adhere cells. Techniques were sought to crosslink aECM proteins in ways that resulted in tunable mechanical properties. Hexamethylene diisocyanate was used to crosslink aECM proteins into uniform, transparent, highly-extensible hydrogel films with low water contents characteristic of native elastin. Their elastic moduli, 0.1--1.1 MPa, depended on crosslinker concentration and aECM protein length, and spanned the observed range of elastin fibers. The suitability of biomaterials implants depends strongly on their susceptibility to proteolytic degradation in vivo. It was shown that small sequence changes in the elastin-like portion of aECM proteins were sufficient to decrease their rate of degradation by elastase sevenfold, illustrating a simple method to tune the protease sensitivity of designed proteins. The effects were seen in both soluble proteins and crosslinked films analyzed by measuring their decrease in elastic modulus during degradation. An aECM protein was examined for its effectiveness as a corneal onlay, i.e., a permanent contact lens. The protein was crosslinked into transparent, elastic, water-rich lenses and was implanted into rabbit corneas. The onlays were stable and well-tolerated, and full re-epithelialization occurred within 4-7 days. Histological examination revealed normal regenerating epithelial cell morphology on the anterior surface, good interfaces between the onlay and surrounding tissue, and only minimal

  18. Cross-linked chitosan thin film coated onto glass plate as an effective adsorbent for adsorption of reactive orange 16.

    PubMed

    Jawad, Ali H; Azharul Islam, Md; Hameed, B H

    2017-02-01

    Fabrication of an immobilized cross-linked chitosan-epichlorohydrine thin film (CLCETF) onto glass plate for adsorption of reactive orange 16 (RO16) dye was successfully studied using the direct casting technique. Adsorption experiments were performed as a function of contact time, initial dye concentration (25mg/L to 350mg/L), and pH (3-11). The adsorption isotherm followed the Langmuir model. The adsorption capacity of CLECTF for RO16 was 356.50mg/g at 27±2°C. The kinetics closely followed the pseudo-second-order model. Results supported the potential use of an immobilized CLECTF as effective adsorbent for the treatment of reactive dye without using filtration process.

  19. Polymeric adsorbent for removing toxic proteins from blood of patients with kidney failure.

    PubMed

    Davankov, V; Pavlova, L; Tsyurupa, M; Brady, J; Balsamo, M; Yousha, E

    2000-02-28

    A hypercrosslinked styrenic polymer with an enhanced proportion of mesopores in the range 2-20 nm has been developed. The principle of the synthesis consists of the suspension polymerization of divinylbenzene (or copolymerization of styrene with divinylbenzene) in the presence of a porogen that is a theta-solvent for polystyrene. On the scale of thermodynamic affinity, theta-solvents occupy a border position between good solvents and precipitating media for the growing polymer chains. In this case, microphase separation takes place during the final stages of the polymerization process. The polymer was shown to adsorb 93-98% of beta2-microglobulin from the blood or plasma of patients with chronic kidney failure. At the same time, large essential proteins, like albumin, are not removed to a significant extent, obviously, due to the size-exclusion effect and the difference in the hydrophobicity of the proteins. By replacing surface exposed pendant vinyl groups of the polymer with hydrophilic functional groups, the material was made hemocompatible, according to the standard battery of biocompatibility tests required by ISO 10993 guidelines. No adverse effects such as fever or hypotension were noted in dogs in direct hemoperfusion experiments with the polymer.

  20. Isolation and characterization of chimeric human Fc-expressing proteins using protein a membrane adsorbers and a streamlined workflow.

    PubMed

    Burdick, Monica M; Reynolds, Nathan M; Martin, Eric W; Hawes, Jacquelyn V; Carlson, Grady E; Cuckler, Chaz M; Bates, Michael C; Barthel, Steven R; Dimitroff, Charles J

    2014-01-08

    Laboratory scale to industrial scale purification of biomolecules from cell culture supernatants and lysed cell solutions can be accomplished using affinity chromatography. While affinity chromatography using porous protein A agarose beads packed in columns is arguably the most common method of laboratory scale isolation of antibodies and recombinant proteins expressing Fc fragments of IgG, it can be a time consuming and expensive process. Time and financial constraints are especially daunting in small basic science labs that must recover hundreds of micrograms to milligram quantities of protein from dilute solutions, yet lack access to high pressure liquid delivery systems and/or personnel with expertise in bioseparations. Moreover, product quantification and characterization may also excessively lengthen processing time over several workdays and inflate expenses (consumables, wages, etc.). Therefore, a fast, inexpensive, yet effective protocol is needed for laboratory scale isolation and characterization of antibodies and other proteins possessing an Fc fragment. To this end, we have devised a protocol that can be completed by limited-experience technical staff in less than 9 hr (roughly one workday) and as quickly as 4 hr, as opposed to traditional methods that demand 20+ work hours. Most required equipment is readily available in standard biomedical science, biochemistry, and (bio)chemical engineering labs, and all reagents are commercially available. To demonstrate this protocol, representative results are presented in which chimeric murine galectin-1 fused to human Fc (Gal-1hFc) from cell culture supernatant was isolated using a protein A membrane adsorber. Purified Gal-1hFc was quantified using an expedited Western blotting analysis procedure and characterized using flow cytometry. The streamlined workflow can be modified for other Fc-expressing proteins, such as antibodies, and/or altered to incorporate alternative quantification and characterization

  1. Protein adsorption resistance of PVP-modified polyurethane film prepared by surface-initiated atom transfer radical polymerization

    NASA Astrophysics Data System (ADS)

    Yuan, Huihui; Qian, Bin; Zhang, Wei; Lan, Minbo

    2016-02-01

    An anti-fouling surface of polyurethane (PU) film grafted with Poly(N-vinylpyrrolidone) (PVP) was prepared through surface-initiated atom transfer radical polymerization (SI-ATRP). And the polymerization time was investigated to obtain PU films with PVP brushes of different lengths. The surface properties and protein adsorption of modified PU films were evaluated. The results showed that the hydrophilicity of PU-PVP films were improved with the increase of polymerization time, which was not positive correlation with the surface roughness due to the brush structure. Additionally, the protein resistance performance was promoted when prolonging the polymerization time. The best antifouling PU-PVP (6.0 h) film reduced the adsoption level of bovine serum albumin (BSA), lysozyme (LYS), and brovin serum fibrinogen (BFG) by 93.4%, 68.3%, 85.6%, respectively, compared to the unmodified PU film. The competitive adsorption of three proteins indicated that LYS preferentially adsorbed on the modified PU film, while BFG had the lowest adsorption selectivity. And the amount of BFG on PU-PVP (6.0 h) film reduced greatly to 0.08 μg/cm2, which was almost one-tenth of its adsorption from the single-protein system. Presented results suggested that both hydrophilicity and surface roughness might be the important factors in all cases of protein adsorption, and the competitive or selective adsorption might be related to the size of the proteins, especially on the non-charged films.

  2. Improved direct electrochemistry for proteins adsorbed on a UV/ozone-treated carbon nanofiber electrode.

    PubMed

    Xue, Qiang; Kato, Dai; Kamata, Tomoyuki; Guo, Qiaohui; You, Tianyan; Niwa, Osamu

    2013-01-01

    We studied the direct electron transfer (DET) of proteins on a carbon nanofiber (CNF) modified carbon film electrode by employing the one-step UV/ozone treatment of CNF. This treatment changed the CNF surface from hydrophobic to hydrophilic because a sufficient quantity of oxygen functional groups was introduced onto the CNF surface. Furthermore, this simple approach increased both the effective surface area and the number of edge-plane defect sites. As a result, the reversibility of redox species, such as ferrocyanide and dopamine, was greatly improved on the treated electrode surface. We obtained on efficient DET of bilirubin oxidase (BOD) and cytochrome c (cyt c) at the treated CNF electrode, which exhibited 38 (for BOD) and 6 (for cyt c) times higher than that at untreated CNF modified electrode. These results indicate that the combination of nanostructured carbon and this UV/ozone treatment process can efficiently create a functionalized surface for the electron transfer of proteins.

  3. Entropy-Driven Conformational Control of α,ω-Difunctional Bidentate-Dithiol Azo-Based Adsorbates Enables the Fabrication of Thermally Stable Surface-Grafted Polymer Films.

    PubMed

    Lee, Han Ju; Jamison, Andrew C; Lee, T Randall

    2016-06-22

    Thermally stable radical initiator monolayers were prepared from uniquely designed α,ω-difunctional adsorbates with bidentate headgroups for the growth of nanoscale polymer films on metal surfaces. The length of the spacer separating the bidentate headgroups was varied to afford 4,4'-(diazene-1,2-diyl)bis(N-(16-(3,5-bis(mercaptomethyl)phenoxy)hexadecyl)-4-cyanopentanamide) (B16), 4,4'-(diazene-1,2-diyl)bis(N-(16-(3,5-bis(mercapto-methyl)phenoxy)decyl)-4-cyanopentanamide) (B10), and 4,4'-(diazene-1,2-diyl)bis(N-(4-(3,5-bis(mercaptomethyl)phenoxy)butyl)-4-cyanopentanamide) (B4). The structural features of the self-assembled monolayers (SAMs) derived from B16, B10, and B4 were characterized by X-ray photoelectron spectroscopy (XPS), ellipsometry, and polarization modulation infrared reflection-absorption spectroscopy (PM-IRRAS) and compared to those derived from an analogous α,ω-difunctional adsorbate with monodentate headgroups, 4,4'-(diazene-1,2-diyl)bis(4-cyano-N-(16-mercaptohexadecyl)pentanamide (M). These studies demonstrate that the conformation (i.e., hairpin vs standing up) of the bidentate initiator adsorbates on gold surfaces was easily controlled by adjusting the concentration of the adsorbates in solution. The results of solution-phase thermal desorption tests revealed that the radical initiator monolayers generated from B16, B10, and B4 exhibit an enhanced thermal stability when compared to those generated from M. Furthermore, a study of the growth of polymer films was performed to evaluate the utility of these new bidentate adsorbate SAMs as film-development platforms for new functional materials and devices. Specifically, surface-grafted polystyrene films were successfully generated from SAMs derived from B16. In contrast, attempts to grow polystyrene films from SAMs derived from M under a variety of analogous conditions were unsuccessful.

  4. A ToF-SIMS and XPS study of protein adsorption and cell attachment across PEG-like plasma polymer films with lateral compositional gradients

    NASA Astrophysics Data System (ADS)

    Menzies, Donna J.; Jasieniak, Marek; Griesser, Hans J.; Forsythe, John S.; Johnson, Graham; McFarland, Gail A.; Muir, Benjamin W.

    2012-12-01

    In this work we report a detailed X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS) study of poly(ethylene glycol) PEG-like chemical gradients deposited via plasma enhanced chemical vapour deposition (PECVD) at two different load powers using diethylene glycol dimethyl ether (DG) as a monomer. Principal component analysis (PCA) was applied to the ToF-SIMS data both before and after protein adsorption on the plasma polymer thin films. Results of the PCA loadings indicated a higher content of hydrocarbon fragments across the higher load power gradient, which adsorbed higher amounts of proteins. Gradients deposited at a lower load power retained a higher degree of monomer like functionality as did the central region directly underneath the knife edge electrode. Analysis of the adsorption of serum proteins (human serum albumin and fetal bovine serum) was monitored across the gradient films and increased with decreasing ether (PEG-like) film chemistries. The effect of protein incubation time on the levels adsorbed fetal bovine serum on the plasma polymer films was critical, with significantly more protein adsorbing after 24 hour incubation times on both gradient films. The attachment of HeLa cells on the gradients appeared to be dictated not only by the surface chemistry, but also by the adsorption of serum proteins. XPS analysis revealed that at surface ether concentrations of less than 70% in the gradient films, significant increases in protein and cell attachment were observed.

  5. Conductivity of copper phthalocyanine-polystyrene composite films in the presence of adsorbed oxygen

    NASA Astrophysics Data System (ADS)

    Pochtennyi, A. E.; Misevich, A. V.; Dolgii, V. K.

    2014-09-01

    The electrical conductivity and adsorption-resistive response to nitrogen dioxide of composite films containing copper phthalocyanine nanoparticles dispersed into the polystyrene matrix are investigated experimentally. The results are analyzed using the two-level model of hopping conductivity. The contributions to the conductivity from intrinsic and impurity localization centers are singled out, and the concentrations of the localization centers in copper phthalocyanines free of impurities as well as the electron localization radii in impurity and intrinsic states are determined.

  6. Kosterlitz-Thouless transition for 4He films adsorbed to rough surfaces.

    PubMed

    Luhman, D R; Hallock, R B

    2004-08-20

    We report the study of adsorption isotherms of 4He on several well characterized rough CaF2 surfaces using a quartz crystal microbalance technique at 1.672 K. The signature of decoupled mass observed on crossing the Kosterlitz-Thouless transition as a function of 4He film thickness decreases and becomes increasingly difficult to identify as the surface roughness is increased. A peak in the dissipation, indicative of the onset of superfluidity, changes little with roughness.

  7. Regulation of protein multipoint adsorption on ion-exchange adsorbent and its application to the purification of macromolecules.

    PubMed

    Huang, Yongdong; Bi, Jingxiu; Zhao, Lan; Ma, Guanghui; Su, Zhiguo

    2010-12-01

    Ion-exchange chromatography (IEC) using commercial ionic absorbents is a widely used technique for protein purification. Protein adsorption onto ion-exchange adsorbents often involves a multipoint adsorption. In IEC of multimeric proteins or "soft" proteins, the intense multipoint binding would make the further desorption difficult, even lead to the destruction of protein structure and the loss of its biological activity. In this paper, DEAE Sepharose FF adsorbents with controllable ligand densities from 0.020 to 0.183 mmol/ml were synthesized, and then the effect of ligand density on the static ion-exchange adsorption of bovine serum albumin (BSA) onto DEAE Sepharose FF was studied by batch adsorption technique. Steric mass-action (SMA) model was employed to analyze the static adsorption behavior. The results showed that the SMA model parameters, equilibrium constant (K(a)), characteristic number of binding sites (υ) and steric factor (σ), increased gradually with ligand density. Thus, it was feasible to regulate BSA multipoint adsorption by modulating the ligand density of ion-exchange adsorbent. Furthermore, IEC of hepatitis B surface antigen (HBsAg) using DEAE Sepharose FF adsorbents with different ligand densities was carried out, and the activity recovery of HBsAg was improved from 42% to 67% when the ligand density was decreased from 0.183 to 0.020 mmol/ml. Taking the activity recovery of HBsAg, the purification factor and the binding capacity into account, DEAE Sepharose FF with a ligand density of 0.041 mmol/ml was most effective for the purification of HBsAg. Such a strategy may also be beneficial for the purification of macromolecules and multimeric proteins.

  8. Effect of adsorbed films on friction of Al2O3-metal systems

    NASA Technical Reports Server (NTRS)

    Pepper, S. V.

    1976-01-01

    The kinetic friction of polycrystalline Al2O3 sliding on Cu, Ni, and Fe in ultrahigh vacuum was studied as a function of the surface chemistry of the metal. Clean metal surfaces were exposed to O2, Cl2, C2H4, and C2H3Cl, and the change in friction due to the adsorbed species was observed. Auger electron spectroscopy assessed the elemental composition of the metal surface. It was found that the systems exposed to Cl2 exhibited low friction, interpreted as the van der Waals force between the Al2O3 and metal chloride. The generation of metal oxide by oxygen exposures resulted in an increase in friction, interpreted as due to strong interfacial bonds established by reaction of metal oxide with Al2O3 to form the complex oxide (spinel). The only effect of C2H4 was to increase the friction of the Fe system, but C2H3Cl exposures decreases friction in both Ni and Fe systems, indicating the dominance of the chlorine over the ethylene complex on the surface

  9. Thick-Film Carbon Dioxide Sensor via Anodic Adsorbate Stripping Technique and Its Structural Dependence.

    PubMed

    Photinon, Kanokorn; Wang, Shih-Han; Liu, Chung-Chiun

    2009-01-01

    A three-electrode based CO(2) sensor was fabricated using thick-film technology. The performance of this sensor was further enhanced by incorporating platinum nanoparticles onto the working electrode surface. An eight-fold increase in the signal output was obtained from the electrode with the platinum nanoparticles. The sensing output was linearly related to the CO(2) presented. Stability measurements demonstrated that the decline of the active surface area and the sensitivity of the sensor were 8% and 13%, respectively, over a two week period of time. The sensor response appeared to be a structural dependence of the crystallographic orientation of platinum electrode.

  10. Bio-photovoltaic conversion device using chlorine-e6 derived from chlorophyll from Spirulina adsorbed on a nanocrystalline TiO2 film electrode.

    PubMed

    Amao, Yutaka; Komori, Tasuku

    2004-03-15

    A bio-photovoltaic conversion device based on dye-sensitised solar cell (DSSC) using the visible light sensitisation of chlorine-e6 (Chl-e6) derived from chlorophyll from Spirulina adsorbed on a nanocrystalline TiO2 film was developed. Form fluorescence spectrum of Chl-e6 adsorbed on a nanocrystalline TiO2 film, the emission of Chl-e6 was effectively quenched by TiO2 nanocrystalline indicating that the effective electron injection from the excited singlet state of Chl-e6 into the conduction band of TiO2 particles occurred. The short-circuit photocurrent density (Isc). the open-circuit photovoltage (Voc). and the fill factor (FF) of solar cell using Chl-e6 adsorbed on a nanocrystalline TiO2 film electrode were estimated to be 0.305 +/- 0.012 mA cm(-2), 426 +/- 10 mV, and 45.0%, respectively. IPCE values were reached a maximum around the wavelength of absorption maximum (7.40% at 400 nm; 1.44% at 514 nm and 2.91% at 670 nm), indicating that the DSSC using visible light sensitisation of nanocrystalline TiO2 film by Chl-e6 was developed.

  11. Infrared spectroscopy of water clusters co-adsorbed with hydrogen molecules on a sodium chloride film

    NASA Astrophysics Data System (ADS)

    Yamakawa, Koichiro; Fukutani, Katsuyuki

    2016-06-01

    Hydrogen gas containing a trace of water vapor was dosed on a vacuum-evaporated sodium chloride film at 13 K, and water clusters formed on the substrate were investigated by infrared absorption spectroscopy. Absorption bands due to (H2O)n clusters with n = 3-6 and an induced absorption band due to hydrogen were clearly observed. With increasing gas dosage, the intensities of the cluster bands increased linearly while the intensity of the hydrogen band was constant. This suggests that the water clusters were formed in two-dimensional matrices of hydrogen. We found that the water clusters did exist on the surface upon heating even after the hydrogen molecules had desorbed. A further rise of the substrate temperature up to 27 K yielded the formation of larger clusters, (H2O)n with n > 6 . We also discuss the origins of the two bands of the trimer in terms of pseudorotation and a metastable isomer.

  12. David Adler Lectureship Award Talk: Friction and energy dissipation mechanisms in adsorbed molecules and molecularly thin films

    NASA Astrophysics Data System (ADS)

    Krim, Jacqueline

    2015-03-01

    Studies of the fundamental origins of friction have undergone rapid progress in recent years, with the development of new experimental and computational techniques for measuring and simulating friction at atomic length and time scales. The increased interest has sparked a variety of discussions and debates concerning the nature of the atomic-scale and quantum mechanisms that dominate the dissipative process by which mechanical energy is transformed into heat. Measurements of the sliding friction of physisorbed monolayers and bilayers can provide information on the relative contributions of these various dissipative mechanisms. Adsorbed films, whether intentionally applied or present as trace levels of physisorbed contaminants, moreover are ubiquitous at virtually all surfaces. As such, they impact a wide range of applications whose progress depends on precise control and/or knowledge of surface diffusion processes. Examples include nanoscale assembly, directed transport of Brownian particles, material flow through restricted geometries such as graphene membranes and molecular sieves, passivation and edge effects in carbon-based lubricants, and the stability of granular materials associated with frictional and frictionless contacts. Work supported by NSFDMR1310456.

  13. Effect of divalent ions on the optical emission behavior of protein thin films

    NASA Astrophysics Data System (ADS)

    Bhowal, Ashim Chandra; Kundu, Sarathi

    2016-05-01

    Photoluminescence behaviors of proteinthin film, bovine serum albumin (BSA) have been studied in the presence of three divalent ions (Mg2+, Ca2+ and Ba2+) at different temperatures using fluorescence spectroscopy. Film thickness and morphology have been studied using atomic force microscopy. Variation of different physicochemical parameters like temperature, solvent polarity, pH, ionic strength, substrate binding etc. can make conformational changes in the protein structure and hence influences the emission behavior.In thin film conformation of BSA, dynamic quenching behavior has beenidentified in the presence of all the three divalent ions at pH≈ 5.5. Depending upon the charge density of the divalent ions interaction with protein molecules modifies and as a result quenching efficiency varies. Also after heat treatment, conformation of the protein molecules changes and as a result the quenching efficiency enhances than that of the unheated films. Studies on such protein-ion interactions and conformational variation may explore various functions of protein when it will adsorb on soft surfaces like membranes, vesicles, etc.

  14. Tribochemical synthesis of nano-lubricant films from adsorbed molecules at sliding solid interface: Tribo-polymers from α-pinene, pinane, and n-decane

    NASA Astrophysics Data System (ADS)

    He, Xin; Barthel, Anthony J.; Kim, Seong H.

    2016-06-01

    The mechanochemical reactions of adsorbed molecules at sliding interfaces were studied for α-pinene (C10H16), pinane (C10H18), and n-decane (C10H22) on a stainless steel substrate surface. During vapor phase lubrication, molecules adsorbed at the sliding interface could be activated by mechanical shear. Under the equilibrium adsorption condition of these molecules, the friction coefficient of sliding steel surfaces was about 0.2 and a polymeric film was tribochemically produced. The synthesis yield of α-pinene tribo-polymers was about twice as much as pinane tribo-polymers. In contrast to these strained bicyclic hydrocarbons, n-decane showed much weaker activity for tribo-polymerization at the same mechanical shear condition. These results suggested that the mechanical shear at tribological interfaces could induce the opening of the strained ring structure of α-pinene and pinane, which leads to polymerization of adsorbed molecules at the sliding track. On a stainless steel surface, such polymerization reactions of adsorbed molecules do not occur under typical surface reaction conditions. The mechanical properties and boundary lubrication efficiency of the produced tribo-polymer films are discussed.

  15. Inhibition of wave-driven two-dimensional turbulence by viscoelastic films of proteins

    NASA Astrophysics Data System (ADS)

    Francois, N.; Xia, H.; Punzmann, H.; Combriat, T.; Shats, M.

    2015-08-01

    To model waves, surface flows, and particle dispersion at the air-water interface one needs to know the essential mechanisms affecting the fluid motion at the surface. We show that a thin film (less than 10-nm thick) of adsorbed protein dramatically affects two-dimensional turbulence generated by Faraday waves at the fluid surface. Extremely low concentrations (≈1 ppm) of soluble proteins form a strong viscoelastic layer which suppresses turbulent diffusion at the surface, changes wave patterns, and shows strong resilience to the wave-induced droplet generation. Surface shear properties of the film play a key role in this phenomenon by inhibiting the creation of vorticity at the surface. The addition of surfactants, on the other hand, destroys the nanolayer and restores the fluid mobility.

  16. The role of polymer nanolayer architecture on the separation performance of anion-exchange membrane adsorbers: I. Protein separations.

    PubMed

    Bhut, Bharat V; Weaver, Justin; Carter, Andrew R; Wickramasinghe, S Ranil; Husson, Scott M

    2011-11-01

    This contribution describes the preparation of strong anion-exchange membranes with higher protein binding capacities than the best commercial resins. Quaternary amine (Q-type) anion-exchange membranes were prepared by grafting polyelectrolyte nanolayers from the surfaces of macroporous membrane supports. A focus of this study was to better understand the role of polymer nanolayer architecture on protein binding. Membranes were prepared with different polymer chain graft densities using a newly developed surface-initiated polymerization protocol designed to provide uniform and variable chain spacing. Bovine serum albumin and immunoglobulin G were used to measure binding capacities of proteins with different size. Dynamic binding capacities of IgG were measured to evaluate the impact of polymer chain density on the accessibility of large size protein to binding sites within the polyelectrolyte nanolayer under flow conditions. The dynamic binding capacity of IgG increased nearly linearly with increasing polymer chain density, which suggests that the spacing between polymer chains is sufficient for IgG to access binding sites all along the grafted polymer chains. Furthermore, the high dynamic binding capacity of IgG (>130 mg/mL) was independent of linear flow velocity, which suggests that the mass transfer of IgG molecules to the binding sites occurs primarily via convection. Overall, this research provides clear evidence that the dynamic binding capacities of large biologics can be higher for well-designed macroporous membrane adsorbers than commercial membrane or resin ion-exchange products. Specifically, using controlled polymerization leads to anion-exchange membrane adsorbers with high binding capacities that are independent of flow rate, enabling high throughput. Results of this work should help to accelerate the broader implementation of membrane adsorbers in bioprocess purification steps.

  17. Physical properties of whey protein--hydroxypropylmethylcellulose blend edible films.

    PubMed

    Brindle, L P; Krochta, J M

    2008-11-01

    The formations of glycerol (Gly)-plasticized whey protein isolate (WPI)-hydroxypropylmethylcellulose (HPMC) films, blended using different combinations and at different conditions, were investigated. The resulting WPI: Gly-HPMC films were analyzed for mechanical properties, oxygen permeability (OP), and water solubility. Differences due to HPMC quantity and blend method were determined via SAS software. While WPI: Gly and HPMC films were transparent, blend films were translucent, indicating some degree of immiscibility and/or WPI-HPMC aggregated domains in the blend films. WPI: Gly-HPMC films were stronger than WPI: Gly films and more flexible and stretchable than HPMC films, with films becoming stiffer, stronger, and less stretchable as the concentration of HPMC increased. However, WPI: Gly-HPMC blended films maintained the same low OP of WPI: Gly films, significantly lower than the OP of HPMC films. Comparison of mechanical properties and OP of films made by heat-denaturing WPI before and after blending with HPMC did not indicate any difference in degree of cross-linking between the methods, while solubility data indicated otherwise. Overall, while adding HPMC to WPI: Gly films had a large effect on the flexibility, strength, stretchability, and water solubility of the film polymeric network, results indicated that HPMC had no effect on OP through the polymer network. WPI-HPMC blend films had a desirable combination of mechanical and oxygen barrier properties, reflecting the combination of hydrogen-bonding, hydrophobic interactions, and disulfide bond cross-linking in the blended polymer network.

  18. Thermoplastic processing of proteins for film formation--a review.

    PubMed

    Hernandez-Izquierdo, V M; Krochta, J M

    2008-03-01

    Increasing interest in high-quality food products with increased shelf life and reduced environmental impact has encouraged the study and development of edible and/or biodegradable polymer films and coatings. Edible films provide the opportunity to effectively control mass transfer among different components in a food or between the food and its surrounding environment. The diversity of proteins that results from an almost limitless number of side-chain amino-acid sequential arrangements allows for a wide range of interactions and chemical reactions to take place as proteins denature and cross-link during heat processing. Proteins such as wheat gluten, corn zein, soy protein, myofibrillar proteins, and whey proteins have been successfully formed into films using thermoplastic processes such as compression molding and extrusion. Thermoplastic processing can result in a highly efficient manufacturing method with commercial potential for large-scale production of edible films due to the low moisture levels, high temperatures, and short times used. Addition of water, glycerol, sorbitol, sucrose, and other plasticizers allows the proteins to undergo the glass transition and facilitates deformation and processability without thermal degradation. Target film variables, important in predicting biopackage performance under various conditions, include mechanical, thermal, barrier, and microstructural properties. Comparisons of film properties should be made with care since results depend on parameters such as film-forming materials, film formulation, fabrication method, operating conditions, testing equipment, and testing conditions. Film applications include their use as wraps, pouches, bags, casings, and sachets to protect foods, reduce waste, and improve package recyclability.

  19. Adsorption of globular proteins on locally planar surfaces. II. Models for the effect of multiple adsorbate conformations on adsorption equilibria and kinetics.

    PubMed Central

    Minton, A P

    1999-01-01

    Equilibrium and kinetic models for nonspecific adsorption of proteins to planar surfaces are presented. These models allow for the possibility of multiple interconvertible surface conformations of adsorbed protein. Steric repulsion resulting in area exclusion by adsorbed molecules is taken into account by treating the adsorbate as a thermodynamically nonideal two-dimensional fluid. In the equilibrium model, the possibility of attractive interactions between adsorbed molecules is taken into account in a limited fashion by permitting one of the adsorbed species to self-associate. Calculated equilibrium adsorption isotherms exhibit apparent high-affinity and low-affinity binding regions, corresponding respectively to adsorption of ligand at low fractional area occupancy in an energetically favorable side-on conformation and conversion at higher fractional area occupancy of the side-on conformation to an entropically favored end-on conformation. Adsorbate self-association may lead to considerable steepening of the adsorption isotherm, compensating to a variable extent for the broadening effect of steric repulsion. Kinetic calculations suggest that in the absence of attractive interactions between adsorbate molecules, the process of adsorption may be highly "stretched" along the time axis, rendering the attainment of adsorption equilibrium in the context of conventional experiments problematic. PMID:9876132

  20. Preparation of Modified Films with Protein from Grouper Fish

    PubMed Central

    Tecante, A.; Granados-Navarrete, S.; Martínez-García, C.

    2016-01-01

    A protein concentrate (PC) was obtained from Grouper fish skin and it was used to prepare films with different amounts of sorbitol and glycerol as plasticizers. The best performing films regarding resistance were then modified with various concentrations of CaCl2, CaSO4 (calcium salts), and glucono-δ-lactone (GDL) with the purpose of improving their mechanical and barrier properties. These films were characterized by determining their mechanical properties and permeability to water vapor and oxygen. Formulations with 5% (w/v) protein and 75% sorbitol and 4% (w/v) protein with a mixture of 15% glycerol and 15% sorbitol produced adequate films. Calcium salts and GDL increased the tensile fracture stress but reduced the fracture strain and decreased water vapor permeability compared with control films. The films prepared represent an attractive alternative for being used as food packaging materials. PMID:27597950

  1. Molecular interactions between proteins and synthetic membrane polymer films

    SciTech Connect

    Pincet, F.; Perez, E.; Belfort, G.

    1995-04-01

    To help understand the effects of protein adsorption on membrane filtration performance, we have measured the molecular interactions between cellulose acetate films and two proteins with different properties (ribonuclease A and human serum albumin) with a surface force apparatus. Comparison of forces between two protein layers with those between a protein layer and a cellulose acetate (CA) film shows that, at high pH, both proteins retained their native conformation on interacting with the CA film while at the isoelectric point (pI) or below the tertiary structure of proteins was disturbed. These measurements provide the first molecular evidence that disruption of protein tertiary structure could be responsible for the reduced permeation flows observed during membrane filtration of protein solutions and suggest that operating at high pH values away from the pI of proteins will reduce such fouling. 60 refs., 9 figs., 5 tabs.

  2. In situ STM imaging of bis-3-sodiumsulfopropyl-disulfide molecules adsorbed on copper film electrodeposited on Pt(111) single crystal electrode.

    PubMed

    Tu, HsinLing; Yen, PoYu; Chen, Sihzih; Yau, ShuehLin; Dow, Wei-Ping; Lee, Yuh-Lang

    2011-06-07

    The adsorption of bis-3-sodiumsulfopropyldi-sulfide (SPS) on metal electrodes in chloride-containing media has been intensively studied to unveil its accelerating effect on Cu electrodeposition. Molecular resolution scanning tunneling microscopy (STM) imaging technique was used in this study to explore the adsorption and decomposition of SPS molecules concurring with the electrodeposition of copper on an ordered Pt(111) electrode in 0.1 M HClO(4) + 1 mM Cu(ClO(4))(2) + 1 mM KCl. Depending on the potential of Pt(111), SPS molecules could react, adsorb, and decompose at chloride-capped Cu films. A submonolayer of Cu adatoms classified as the underpotential deposition (UPD) layer at 0.4 V (vs Ag/AgCl) was completely displaced by SPS molecules, possibly occurring via RSSR (SPS) + Cl-Cu-Pt → RS(-)-Pt(+) + RS(-) (MPS) + Cu(2+) + Cl(-), where MPS is 3-mercaptopropanesulfonate. By contrast, at 0.2 V, where a full monolayer of Cu was presumed to be deposited, SPS molecules were adsorbed in local (4 × 4) structures at the lower ends of step ledges. Bulk Cu deposition driven by a small overpotential (η < 50 mV) proceeded slowly to yield an atomically smooth Cu deposit at the very beginning (<5 layers). On a bilayer Cu deposit, the chloride adlayer was still adsorbed to afford SPS admolecules arranged in a unique 1D striped phase. SPS molecules could decompose into MPS upon further Cu deposition, as a (2 × 2)-MPS structure was observed with prolonged in situ STM imaging. It was possible to visualize either SPS admolecules in the upper plane or chloride adlayer sitting underneath upon switching the imaging conditions. Overall, this study established a MPS molecular film adsorbed to the chloride adlayer sitting atop the Cu deposit.

  3. Development of a novel polystyrene/metal-organic framework-199 electrospun nanofiber adsorbent for thin film microextraction of aldehydes in human urine.

    PubMed

    Liu, Feilong; Xu, Hui

    2017-01-01

    In this work, electrospun polystyrene/metal-organic frameworks-199 (PS/MOF-199) nanofiber film was synthesized and investigated as a novel adsorbent for thin film microextraction (TFME) of aldehydes in human urine. Some properties of the prepared PS/MOF-199 nanofiber film, including morphology, structure, wettability, solvent stability and extraction performance were studied systematically. Porous fibrous structure, large surface area, good stability, strong hydrophobicity and excellent extraction efficiency were obtained for the film. Based on the PS/MOF-199 film, a thin film microextraction-high performance liquid chromatography (TFME-HPLC) method was developed, and the experimental parameters that affected the extraction and desorption were optimized. Under the optimal conditions, the limits of detection (LODs) were in the range of 4.2-17.3nmolL(-1) for the analysis of six aldehydes. Good linearity was achieved with correlation coefficients (R(2)) being lager than 0.9943. Satisfactory recovery (82-112%) and acceptable reproducibility (relative standard deviation: 2.1-13.3%) were also obtained for the method. The developed TFME-HPLC method has been successfully applied to the analysis of aldehyde metabolites in the urine samples of lung cancer patients and healthy people. The method possesses the advantages of simplicity, rapidity, cost-effective, sensitivity and non-invasion, it provides an alternative tool for the determination of aldehydes in complex sample matrices.

  4. Protein immobilization and fluorescence quenching on polydopamine thin films.

    PubMed

    Chen, Daqun; Zhao, Lei; Hu, Weihua

    2016-09-01

    Mussel inspired polydopamine (PDA) film has attracted great interest as a versatile functional coating for biomolecule immobilization in various bio-related devices. However, the details regarding the interaction between a protein and PDA film remain unclear. Particularly, there is very limited knowledge regarding the protein immobilization on PDA film, even though it is of essential importance in various fields. The situation is even more complicated if considering the fact that quite a number of approaches (e.g., different oxidizing reagent, buffer pH, grown time, grown media, etc.) have been developed to grow PDA films. In this work, protein attachment on PDA film was systematically investigated by using the real-time and label-free surface plasmon resonance (SPR) technique. The kinetics of protein-PDA interaction was explored and the influence of buffer pH and deposition media on the protein attachment was studied. Fluorescent protein microarray was further printed on PDA-coated glass slides for quantitative investigations and together with SPR data, the interesting fluorescence quenching phenomenon of PDA film was revealed. This work may deepen our understanding on the PDA-protein interaction and offer a valuable guide for efficient protein attachment on PDA film in various bio-related applications.

  5. Affinity binding of antibodies to supermacroporous cryogel adsorbents with immobilized protein A for removal of anthrax toxin protective antigen.

    PubMed

    Ingavle, Ganesh C; Baillie, Les W J; Zheng, Yishan; Lis, Elzbieta K; Savina, Irina N; Howell, Carol A; Mikhalovsky, Sergey V; Sandeman, Susan R

    2015-05-01

    Polymeric cryogels are efficient carriers for the immobilization of biomolecules because of their unique macroporous structure, permeability, mechanical stability and different surface chemical functionalities. The aim of the study was to demonstrate the potential use of macroporous monolithic cryogels for biotoxin removal using anthrax toxin protective antigen (PA), the central cell-binding component of the anthrax exotoxins, and covalent immobilization of monoclonal antibodies. The affinity ligand (protein A) was chemically coupled to the reactive hydroxyl and epoxy-derivatized monolithic cryogels and the binding efficiencies of protein A, monoclonal antibodies to the cryogel column were determined. Our results show differences in the binding capacity of protein A as well as monoclonal antibodies to the cryogel adsorbents caused by ligand concentrations, physical properties and morphology of surface matrices. The cytotoxicity potential of the cryogels was determined by an in vitro viability assay using V79 lung fibroblast as a model cell and the results reveal that the cryogels are non-cytotoxic. Finally, the adsorptive capacities of PA from phosphate buffered saline (PBS) were evaluated towards a non-glycosylated, plant-derived human monoclonal antibody (PANG) and a glycosylated human monoclonal antibody (Valortim(®)), both of which were covalently attached via protein A immobilization. Optimal binding capacities of 108 and 117 mg/g of antibody to the adsorbent were observed for PANG attached poly(acrylamide-allyl glycidyl ether) [poly(AAm-AGE)] and Valortim(®) attached poly(AAm-AGE) cryogels, respectively, This indicated that glycosylation status of Valortim(®) antibody could significantly increase (8%) its binding capacity relative to the PANG antibody on poly(AAm-AGE)-protien-A column (p < 0.05). The amounts of PA which remained in the solution after passing PA spiked PBS through PANG or Valortim bound poly(AAm-AGE) cryogel were significantly (p < 0

  6. Electrostatically-controlled protein adsorption onto lipid bilayer: modeling adsorbate aggregation behavior.

    PubMed

    Trusova, Valeriya M; Gorbenko, Galyna P

    2008-03-01

    Using adsorption models based on scaled particle (SPT) and double layer theories the electrostatically-controlled protein adsorption onto membrane surface has been simulated for non-associating and self-associating ligands. The binding isotherms of monomeric and oligomeric protein species have been calculated over a range of variable parameters including lipid and protein concentrations, protein and membrane charges, pH and ionic strength. Adsorption behavior of monomers appeared to be the most sensitive to the changes in the protein aggregation state. The hallmarks of the protein oligomerization are identified. The practical guides for optimal design of binding experiments focused on obtaining proofs of protein self-association are suggested.

  7. Synovial fluid lubrication of artificial joints: protein film formation and composition.

    PubMed

    Fan, Jingyun; Myant, Connor; Underwood, Richard; Cann, Philippa

    2012-01-01

    Despite design improvements, wear of artificial implants remains a serious health issue particularly for Metal-on-Metal (MoM) hips where the formation of metallic wear debris has been linked to adverse tissue response. Clearly it is important to understand the fundamental lubrication mechanisms which control the wear process. It is usually assumed that MoM hips operate in the ElastoHydrodynamic Lubrication (EHL) regime where film formation is governed by the bulk fluid viscosity; however there is little experimental evidence of this. The current paper critically examines synovial fluid lubrication mechanisms and the effect of synovial fluid chemistry. Two composition parameters were chosen; protein content and pH, both of which are known to change in diseased or post-operative synovial fluid. Film thickness and wear tests were carried out for a series of model synovial fluid solutions. Two distinct film formation mechanisms were identified; an adsorbed surface film and a high-viscosity gel. The entrainment of this gel controls film formation particularly at low speeds. However wear of the femoral head still occurs and this is thought to be due primarily to a tribo-corrosion mechanisms. The implications of this new lubrication mechanism and the effect of different synovial fluid chemistries are examined. One important conclusion is that patient synovial fluid chemistry plays an important role in determining implant wear and the likelihood of failure.

  8. IgG adsorption on a new protein A adsorbent based on macroporous hydrophilic polymers. I. Adsorption equilibrium and kinetics.

    PubMed

    Perez-Almodovar, Ernie X; Carta, Giorgio

    2009-11-20

    Experimental determination and modeling of IgG binding on a new protein A adsorbent based on a macroporous resin were performed. The new adsorbent consists of polymeric beads based on hydrophilic acrylamido and vinyl monomers with a pore structure optimized to allow favorable interactions of IgG with recombinant protein A coupled to the resin. The particles have average diameter of 57 microm and a narrow particle size distribution. The IgG adsorption equilibrium capacity is 46 mg/cm(3) and the effective pore diffusivity determined from pulse response experiments for non-binding conditions is 8.0 x 10(-8) cm(2)/s. The IgG adsorption kinetics can be described with the same effective diffusivity by taking into account a heterogeneous binding mechanism with fast binding sites, for which adsorption is completely diffusion controlled, and slow binding sites for which adsorption is controlled by the binding kinetics. As a result of this mechanism, the breakthrough curve exhibits a tailing behavior, which appears to be associated with the slow binding sites. A detailed rate model taking into account intraparticle diffusion and binding kinetics is developed and is found capable of predicting both batch adsorption and breakthrough behavior over an ample range of experimental conditions. The corresponding effective diffusivity is independent of protein concentration in solution over the range 0.2-2 mg/cm(3) and of protein binding as a result of the large pore size of the support matrix. Overall, the small particle size and low diffusional hindrance allow capture of IgG with short residence times while attaining substantial dynamic binding capacities.

  9. Adsorption of proteins at physiological concentrations on pegylated surfaces and the compatibilizing role of adsorbed albumin with respect to other proteins according to optical waveguide lightmode spectroscopy (OWLS).

    PubMed

    Leclercq, Laurent; Modena, Enrico; Vert, Michel

    2013-01-01

    In literature, contacts between pegylated compounds and blood proteins are generally discussed in terms of excluded volume-related repulsions although adsorption and compatibility have been reported for some of these proteins occasionally. The major problem to investigate the behavior of blood in contact with pegylated surfaces is the complexity of the medium and especially the presence of albumin in large excess. In a model approach, optical waveguide lightmode spectroscopy (OWLS) was used to monitor the fate of albumin, fibrinogen, and γ-globulins at physiological concentrations in pH = 7.4 isotonic HEPES buffer after contact with SiTiO2 chips coated with diblock poly(DL-lactic acid)-block-poly(ethylene oxide)s and triblock poly(DL-lactic acid)-block-poly(ethylene oxide)-block-poly(DL-lactic acid) copolymers. Corresponding homopolymers were used as controls. The three protein systems were investigated separately, as a mixture and when added successively according to different orders of addition. OWLS gave access to the mass and the thickness of adhering protein layers that resist washing with HEPES buffer. Protein depositions were detected regardless of the presence of poly(ethylene glycol) segments on surfaces. Adsorption depended on the protein, on the surface and also on the presence of the other proteins. Unexpectedly any surface coated with a layer of adsorbed albumin prevented deposition of other proteins, including albumin itself. This outstanding finding suggests that it was the presence of albumin adsorbed on a surface, pegylated or not, that made that surface compatible with other proteins. As a consequence, dipping a device to be in contact with the blood of a patient in a solution of albumin could be a very simple means to avoid further protein deposition and maybe platelets adhesion after in vivo implantation.

  10. Theory and applications of refractive index-based optical microscopy to measure protein mass transfer in spherical adsorbent particles.

    PubMed

    Bankston, Theresa E; Stone, Melani C; Carta, Giorgio

    2008-04-25

    This work provides the theoretical foundation and a range of practical application examples of a recently developed method to measure protein mass transfer in adsorbent particles using refractive index-based optical microscopy. A ray-theoretic approach is first used to predict the behavior of light traveling through a particle during transient protein adsorption. When the protein concentration gradient in the particle is sharp, resulting in a steep refractive index gradient, the rays bend and intersect, thereby concentrating light in a sharp ring that marks the position of the adsorption front. This behavior is observed when mass transfer is dominated by pore diffusion and the adsorption isotherm is highly favorable. Applications to protein cation-exchange, hydrophobic interaction, and affinity adsorption are then considered using, as examples, the three commercial, agarose-based stationary phases SP-Sepharose-FF, Butyl Sepharose 4FF, and MabSelect. In all three cases, the method provides results that are consistent with measurements based on batch adsorption and previously published data confirming its utility for the determination of protein mass transfer kinetics under a broad range of practically relevant conditions.

  11. Heavy metal-binding proteins from metal-stimulated bacteria as a novel adsorbent for metal removal technology.

    PubMed

    Sano, D; Myojo, K; Omura, T

    2006-01-01

    Water pollution with toxic heavy metals is of growing concern because heavy metals could bring about serious problems for not only ecosystems in the water environment but also human health. Some metal removal technologies have been in practical use, but much energy and troublesome treatments for chemical wastes are required to operate these conventional technologies. In this study, heavy metal-binding proteins (HMBPs) were obtained from metal-stimulated activated sludge culture with affinity chromatography using copper ion as a ligand. Two-dimensional electrophoresis revealed that a number of proteins in activated sludge culture were recovered as HMBPs for copper ion. N-termini of five HMBPs were determined, and two of them were found to be newly discovered proteins for which no amino acid sequences in protein databases were retrieved at more than 80% identities. Metal-coordinating amino acids occupied 38% of residues in one of the N-terminal sequences of the newly discovered HMBPs. Since these HMBPs were expected to be stable under conditions of water and wastewater treatments, it would be possible to utilize HMBPs as novel adsorbents for heavy metal removal if mass volume of HMBPs can be obtained with protein cloning techniques.

  12. Sum Frequency Generation Vibrational Spectroscopy of Adsorbed Amino Acids, Peptides and Proteins of Hydrophilic and Hydrophobic Solid-Water Interfaces

    SciTech Connect

    Holinga IV, George Joseph

    2010-09-01

    Sum frequency generation (SFG) vibrational spectroscopy was used to investigate the interfacial properties of several amino acids, peptides, and proteins adsorbed at the hydrophilic polystyrene solid-liquid and the hydrophobic silica solid-liquid interfaces. The influence of experimental geometry on the sensitivity and resolution of the SFG vibrational spectroscopy technique was investigated both theoretically and experimentally. SFG was implemented to investigate the adsorption and organization of eight individual amino acids at model hydrophilic and hydrophobic surfaces under physiological conditions. Biointerface studies were conducted using a combination of SFG and quartz crystal microbalance (QCM) comparing the interfacial structure and concentration of two amino acids and their corresponding homopeptides at two model liquid-solid interfaces as a function of their concentration in aqueous solutions. The influence of temperature, concentration, equilibration time, and electrical bias on the extent of adsorption and interfacial structure of biomolecules were explored at the liquid-solid interface via QCM and SFG. QCM was utilized to quantify the biological activity of heparin functionalized surfaces. A novel optical parametric amplifier was developed and utilized in SFG experiments to investigate the secondary structure of an adsorbed model peptide at the solid-liquid interface.

  13. Effect of surface microstructure and wettability on plasma protein adsorption to ZnO thin films prepared at different RF powers.

    PubMed

    Huang, Zhan-Yun; Chen, Min; Pan, Shi-Rong; Chen, Di-Hu

    2010-10-01

    In this paper, the adsorption behavior of plasma proteins on the surface of ZnO thin films prepared by radio frequency (RF) sputtering under different sputtering powers was studied. The microstructures and surface properties of the ZnO thin films were investigated by x-ray diffraction (XRD), scanning electron microscopy (SEM), UV-visible optical absorption spectroscopy and contact angle techniques. The results show that the ZnO thin films have better orientation of the (0 0 2) peak with increasing RF power, especially at around 160 W, and the optical band gap of the ZnO films varies from 3.2 to 3.4 eV. The contact angle test carried out by the sessile drop technique denoted a hydrophobic surface of the ZnO films, and the surface energy and adhesive work of the ZnO thin films decreased with increasing sputtering power. The amounts of human fibrinogen (HFG) and human serum albumin (HSA) adsorbing on the ZnO films and reference samples were determined by using enzyme-linked immunosorbent assay (ELISA). The results show that fewer plasma proteins and a smaller HFG/HSA ratio adsorb on the ZnO thin films' surface.

  14. Probing the binding affinity of plasma proteins adsorbed on Au nanoparticles.

    PubMed

    Zhang, Xiaoning; Zhang, Junting; Zhang, Fan; Yu, Shaoning

    2017-04-06

    Nanoparticle (NP) surfaces are modified immediately by the adsorption of proteins when exposed to human blood, leading to the formation of a protein corona. The adsorption of serum proteins is the key process for exploring the bioapplication and biosafety of NPs. In this study, NP-protein binding affinity (Ka) was investigated. Some serum proteins, such as human serum albumin (HSA), trypsin (TRP), hemoglobin (Hb), myoglobin (MB), immunoglobulin G (IgG), carbonic anhydrase (CA), fibrinogen (FIB), chymotrypsin and r-globulin, were used with gold nanoparticles (AuNPs) to address binding affinity according to isothermal titration calorimetry (ITC) combined with dynamic light scattering (DLS) and fluorescence quenching. The NP protein binding affinities determined by the two methods were in agreement, and depended on the protein properties and size of the NPs. The two methods are convenient, and the results are highly comparable. These methods can be extended to determine the binding affinity of NP protein interactions. The adsorption of proteins upon the AuNP surface is a complex process and depends on several factors, but the binding affinities are higher for proteins with more cysteine residues located on the surface.

  15. Assessment of fibronectin conformation adsorbed to polytetrafluoroethylene surfaces from serum protein mixtures and correlation to support of cell attachment in culture.

    PubMed

    Grainger, David W; Pavon-Djavid, Graciella; Migonney, Veronique; Josefowicz, Marcel

    2003-01-01

    Surfaces of polytetrafluoroethylene (PTFE) were exposed to buffered aqueous solutions containing radio-labeled human fibronectin ([125I]Fn), Fn/bovine serum albumin (BSA) binary mixtures of various ratios or whole human plasma dilutions for 1 h. Total adsorbed Fn and albumin adsorption following rinsing was quantified on this surface. 125I-labeled monoclonal antibodies against either the tenth type-III Fn repeat unit (containing the cell-binding RGDS integrin recognition motif) or the Fn amino-terminal domain were used to probe the accessibility of each of these respective Fn regions post-adsorption. Human umbilical vein endothelial cells (HUVECs) were cultured on PTFE surfaces pre-exposed to each of these protein adsorption conditions and compared to identical conditions on tissue culture polystyrene (TCPS). Fn adsorption to PTFE is dependent upon the concentration of albumin co-adsorbing from solution: albumin out-competes Fn for PTFE surface sites even at non-physiological Fn/HSA ratios 10-100-fold biased in Fn. Antibodies against Fn do not readily recognize Fn adsorbed on PTFE as the HSA co-adsorption concentration in either binary mixtures or in plasma increases, indicating albumin masking of adsorbed Fn. At Fn/HSA ratios rich in Fn (1:1, 1:100), albumin co-adsorption actually improves anti-Fn antibody recognition of adsorbed Fn. HUVEC attachment efficiency to PTFE after protein adsorption correlates with amounts of Fn adsorbed and levels of anti-Fn antibody recognition of Fn on PTFE, linking cell attachment to integrin recognition of both adsorbed Fn density and Fn adsorbed conformation on PTFE surfaces.

  16. The Development of an Edible Peanut Protein Film

    NASA Technical Reports Server (NTRS)

    Patrick, N.; Jones, G.; Aglan, H.; Lu, J.

    1998-01-01

    The peanut is one of the crops chosen for use in NASA's Advanced Life Support Program (ALS). The peanut is a source of both oil and protein. After oil is extracted from the peanut, a protein rich flour remains. An edible peanut protein film is one use for this flour. Two types of film are developed for this study, one set of film contains 10% fat while the other set contains no fat. For film without fat the defatting of the peanut by the Soxhlet method is the first step in the manufacturing process of the film. Secondly, the protein is precipitated at its isoelectric point (pH 4.5) and centrifuged to separate the protein from the non-protein. After freeze-drying the protein it is milled in a ceramic ball mill to decrease particle size and sifted through a series of sieves to determine particle size distribution. Those particles retained on the 100 mesh sieves are utilized for film formation. Larger particles are re-ground and sifted. Five grams of protein is mixed with 50 mL of distilled water, 70 mL of 80% ethanol, 15 mL of 6N ammonium hydroxide and a plasticizer. This mixture is heated for 30 minutes until the temperature reaches 70 C. The mixture is then poured onto a level Teflon coated glass surface. After allowing the film to form overnight under a ventilation hood, it is manually removed from the plate. The processes and methods adopted have created flexible films of uniform thickness that are free of air bubbles. Thickness of films made from defatted peanut protein and partially defatted peanut protein were 0.10 Lm and 0.13 Lm respectively. Films with natural peanut fat are approximately three times as flexible and almost four times as strong as the films made without fat. Further research will be performed to evaluate its mechanical properties. This paper will greatly contribute to food preservation and waste management. Potential applications of this film are edible/biodegradable containers, wrapping for food preservation (against water, oxygen and oil

  17. Selective binding of single-stranded DNA-binding proteins onto DNA molecules adsorbed on single-walled carbon nanotubes.

    PubMed

    Nii, Daisuke; Hayashida, Takuya; Yamaguchi, Yuuki; Ikawa, Shukuko; Shibata, Takehiko; Umemura, Kazuo

    2014-09-01

    Single-stranded DNA-binding (SSB) proteins were treated with hybrids of DNA and single-walled carbon nanotubes (SWNTs) to examine the biological function of the DNA molecules adsorbed on the SWNT surface. When single-stranded DNA (ssDNA) was used for the hybridization, significant binding of the SSB molecules to the ssDNA-SWNT hybrids was observed by using atomic force microscopy (AFM) and agarose gel electrophoresis. When double-stranded DNA (dsDNA) was used, the SSB molecules did not bind to the dsDNA-SWNT hybrids in most of the conditions that we evaluated. A specifically modified electrophoresis procedure was used to monitor the locations of the DNA, SSB, and SWNT molecules. Our results clearly showed that ssDNA/dsDNA molecules on the SWNT surfaces retained their single-stranded/double-stranded structures.

  18. Fabrication and characterization of macroporous epichlorohydrin cross-linked alginate beads as protein adsorbent.

    PubMed

    Zhang, Weican; Ji, Xiaofei; Sun, Caiyun; Lu, Xuemei

    2013-01-01

    Porous epichlorohydrin cross-linked alginate beads (ECAB) were prepared by the following method. Na-alginate solution containing Na2SO4 was introduced dropwise into CaCl2 solution to simultaneously form CaSO4 precipitate and Ca-alginate gel beads. The resultant beads were cross-linked with epichlorohydrin and then thoroughly washed with ethylenediamine tetraacetic acid (EDTA) solution to remove CaSO4. The structural features of porous ECAB were assessed with scanning electron microscopy (SEM) and experiments on water content and adsorption of bovine serum albumin (BSA). The results showed that macroporous ECAB can be obtained when the mass ratio of sodium sulfate to sodium alginate is 4:1. The adsorption behavior of the macroporous ECAB was well described by the Langmuir isotherm with maximum adsorption capacity equal to 740 mg BSA/g dry weight in 50 mM Na2HPO4-citric acid buffer (pH 4.0). BSA was more effectively adsorbed by macroporous ECAB at around pH 3 and the mechanism of the adsorption of BSA to the ECAB was ion exchange. Finally, experiments of a concentration of 1 mg/mL BSA using macroporous ECAB were performed.

  19. In situ x-ray photoelectron spectroscopic and density-functional studies of Si atoms adsorbed on a C60 film.

    PubMed

    Onoe, Jun; Nakao, Aiko; Hara, Toshiki

    2004-12-08

    The interaction between C(60) and Si atoms has been investigated for Si atoms adsorbed on a C(60) film using in situ x-ray photoelectron spectroscopy (XPS) and density-functional (DFT) calculations. Analysis of the Si 2p core peak identified three kinds of Si atoms adsorbed on the film: silicon suboxides (SiO(x)), bulk Si crystal, and silicon atoms bound to C(60). Based on the atomic percent ratio of silicon to carbon, we estimated that there was approximately one Si atom bound to each C(60) molecule. The Si 2p peak due to the Si-C(60) interaction demonstrated that a charge transfer from the Si atom to the C(60) molecule takes place at room temperature, which is much lower than the temperature of 670 K at which the charge transfer was observed for C(60) adsorbed on Si(001) and (111) clean surfaces [Sakamoto et al., Phys. Rev. B 60, 2579 (1999)]. The number of electrons transferred between the C(60) molecule and Si atom was estimated to be 0.59 based on XPS results, which is in good agreement with the DFT result of 0.63 for a C(60)Si with C(2v) symmetry used as a model cluster. Furthermore, the shift in binding energy of both the Si 2p and C 1s core peaks before and after Si-atom deposition was experimentally obtained to be +2.0 and -0.4 eV, respectively. The C(60)Si model cluster provides the shift of +2.13 eV for the Si 2p core peak and of -0.28 eV for the C 1s core peak, which are well corresponding to those experimental results. The covalency of the Si-C(60) interaction was also discussed in terms of Mulliken overlap population between them.

  20. Application of an Antimicrobial Protein Film in Beef Patties Packaging

    PubMed Central

    2015-01-01

    This study was performed to apply a protein film containing a natural antimicrobial compound to meat packaging and determine quality change of meat during storage. Proteins obtained from the by-products of food processing have been utilized as biodegradable film sources. Porcine meat and bone meal (MBM) is obtained during meat processing, and proteins from the MBM can be extracted and used as a film base material. Previously, an antimicrobial MBM film containing coriander oil (CO) was prepared and its physical properties and antimicrobial activity were characterized. In this study, the antimicrobial MBM-CO film was applied to beef patties packaging, and the microbial population and the degree of lipid oxidation were determined during storage at 4℃ for 15 d. The population of inoculated E. coli O157:H7 in the samples wrapped with the MBM-CO film was 6.78 log colony forming unit (CFU)/g after 15 d of storage, whereas the control had 8.05 Log CFU/g, thus reducing the microbial population by 1.29 Log CFU/g. In addition, retardation of lipid oxidation in the patties was observed during storage for the samples packaged by the MBM-CO film, compared with the control samples. These results suggest that the MBM-CO film can be useful for enhancing the quality of beef patties during storage. PMID:26761888

  1. Effect of protein concentrations on the properties of fish myofibrillar protein based film compared with PVC film.

    PubMed

    Kaewprachu, Pimonpan; Osako, Kazufumi; Benjakul, Soottawat; Rawdkuen, Saroat

    2016-04-01

    The effect of protein concentrations on the properties of fish myofibrillar protein film (FMP) were investigated and compared with commercial wrap film (polyvinyl chloride; PVC). FMP (2 %, w/v) showed the highest mechanical properties [tensile strength: 4.38 MPa and elongation at break: 133.05 %], and water vapor permeability [2.81 × 10(-10) g m(-1) s(-1) Pa(-1)]. FMP contained high molecular weight cross-links, resulting in complex film network, as indicated by lower film solubility (19-22 %) and protein solubility (0.6-1.3 %). FMP showed excellent barrier properties to UV light at the wavelength of 200-280 nm. FMP had the thickness [0.007-0.032 mm], color attributes and transparency similar to PVC film [thickness: 0.010 mm]. Therefore, protein concentration majority influenced the properties of develop FMP. The protein content of 1 % (w/v) had potential to be developed the biodegradable film with comparable properties to the commercial wrap film.

  2. Heterogeneous patterns on block copolymer thin film via solvent annealing: Effect on protein adsorption

    NASA Astrophysics Data System (ADS)

    Shen, Lei; Zhu, Jintao; Liang, Haojun

    2015-03-01

    Heterogeneous patterns consisting of nanometer-scaled hydrophobic/hydrophilic domains were generated by self-assembly of poly(styrene)-block-poly(2-hydroxyethyl methacrylate) (PS-b-PHEMA) block copolymer thin film. The effect of the heterogeneity of the polymer film surface on the nonspecific adsorption of the protein human plasma fibrinogen (FBN, 5.0 × 5.0 × 47.5 nm3) was investigated. The kinetics of the FBN adsorption varies from a single-component Langmuir model on homogeneous hydrophilic PHEMA to a two-stage spreading relaxation model on homogeneous hydrophobic PS surface. On a heterogeneous PS-b-PHEMA surface with majority PS part, the initial FBN adsorption rate remains the same as that on the homogeneous PS surface. However, hydrophilic PHEMA microdomains on the heterogeneous surface slow down the second spreading stage of the FBN adsorption process, leading to a surface excess of adsorbed FBN molecules less than the presumed one simply calculated as adsorption onto multiple domains. Importantly, when the PS-b-PHEMA surface is annealed to form minority domelike PS domains (diameter: ˜50-100 nm) surrounded by a majority PHEMA matrix, such surface morphology proves to be strongly protein-repulsive. These interesting findings can be attributed to the enhancement of the spread FBN molecule in a mobile state by the heterogeneity of polymer film surface before irreversible adsorption occurs.

  3. A theoretical and experimental approach toward the development of affinity adsorbents for GFP and GFP-fusion proteins purification.

    PubMed

    Fernandes, Cláudia S M; Pina, Ana Sofia; Dias, Ana M G C; Branco, Ricardo J F; Roque, Ana Cecília Afonso

    2014-09-30

    The green fluorescent protein (GFP) is widely employed to report on a variety of molecular phenomena, but its selective recovery is hampered by the lack of a low-cost and robust purification alternative. This work reports an integrated approach combining rational design and experimental validation toward the optimization of a small fully-synthetic ligand for GFP purification. A total of 56 affinity ligands based on a first-generation lead structure were rationally designed through molecular modeling protocols. The library of ligands was further synthesized by solid-phase combinatorial methods based on the Ugi reaction and screened against Escherichia coli extracts containing GFP. Ligands A4C2, A5C5 and A5C6 emerged as the new lead structures based on the high estimated theoretical affinity constants and the high GFP binding percentages and enrichment factors. The elution of GFP from these adsorbents was further characterized, where the best compromise between mild elution conditions, yield and purity was found for ligands A5C5 and A5C6. These were tested for purifying a model GFP-fusion protein, where ligand A5C5 yielded higher protein recovery and purity. The molecular interactions between the lead ligands and GFP were further assessed by molecular dynamics simulations, showing a wide range of potential hydrophobic and hydrogen-bond interactions.

  4. Capillary electrophoresis-mass spectrometry of basic proteins using a new physically adsorbed polymer coating. Some applications in food analysis.

    PubMed

    Simó, Carolina; Elvira, Carlos; González, Nieves; San Román, J; Barbas, Coral; Cifuentes, Alejandro

    2004-07-01

    A new physically adsorbed capillary coating for capillary electrophoresis-mass spectrometry (CE-MS) of basic proteins is presented, which is easily obtained by flushing the capillary with a polymer aqueous solution for two min. This coating significantly reduces the electrostatic adsorption of a group of basic proteins (i.e., cytochrome c, lysozyme, and ribonuclease A) onto the capillary wall allowing their analysis by CE-MS. The coating protocol is compatible with electrospray inonization (ESI)-MS via the reproducible separation of the standard basic proteins (%RSD values (n = 5) < 1% for analysis time reproducibility and < 5% for peak heights, measured from the total ion electropherograms (TIEs) within the same day). The LODs determined using cytochrome c with total ion current and extracted ion current defection were 24.5 and 2.9 fmol, respectively. Using this new coating lysozymes from chicken and turkey egg white could be easily distinguished by CE-MS, demonstrating the usefulness of this method to differentiate animal species. Even after sterilization at 120 degrees C for 30 min, lysozyme could be detected, as well as in wines at concentrations much lower than the limit marked by the EC Commission Regulation. Adulteration of minced meat with 5% of egg-white could also be analysed by our CE-MS protocol.

  5. Elastic properties of protein functionalized nanoporous polymer films

    SciTech Connect

    Charles T. Black; Wang, Haoyu; Akcora, Pinar

    2015-12-16

    Retaining the conformational structure and bioactivity of immobilized proteins is important for biosensor designs and drug delivery systems. Confined environments often lead to changes in conformation and functions of proteins. In this study, lysozyme is chemically tethered into nanopores of polystyrene thin films, and submicron pores in poly(methyl methacrylate) films are functionalized with streptavidin. Nanoindentation experiments show that stiffness of streptavidin increases with decreasing submicron pore sizes. Lysozymes in polystyrene nanopores are found to behave stiffer than the submicron pore sizes and still retain their specific bioactivity relative to the proteins on flat surfaces. Lastly, our results show that protein functionalized ordered nanoporous polystyrene/poly(methyl methacrylate) films present heterogeneous elasticity and can be used to study interactions between free proteins and designed surfaces.

  6. Analysis of diffusion models for protein adsorption to porous anion-exchange adsorbent.

    PubMed

    Chen, Wei-Dong; Dong, Xiao-Yan; Sun, Yan

    2002-07-12

    The ion-exchange adsorption kinetics of bovine serum albumin (BSA) and gamma-globulin to an anion exchanger, DEAE Spherodex M, has been studied by batch adsorption experiments. Various diffusion models, that is, pore diffusion, surface diffusion, homogeneous diffusion and parallel diffusion models, are analyzed for their suitabilities to depict the adsorption kinetics. Protein diffusivities are estimated by matching the models with the experimental data. The dependence of the diffusivities on initial protein concentration is observed and discussed. The adsorption isotherm of BSA is nearly rectangular, so there is little surface diffusion. As a result, the surface and homogeneous diffusion models do not fit to the kinetic data of BSA adsorption. The adsorption isotherm of gamma-globulin is less favorable, and the surface diffusion contributes greatly to the mass transport. Consequently, both the surface and homogeneous diffusion models fit to the kinetic data of gamma-globulin well. The adsorption kinetics of BSA and gamma-globulin can be very well fitted by parallel diffusion model, because the model reflects correctly the intraparticle mass transfer mechanism. In addition, for both the favorably bound proteins, the pore diffusion model fits the adsorption kinetics reasonably well. The results here indicate that the pore diffusion model can be used as a good approximate to depict protein adsorption kinetics for protein adsorption systems from rectangular to linear isotherms.

  7. Adsorbent phosphates

    NASA Technical Reports Server (NTRS)

    Watanabe, S.

    1983-01-01

    An adsorbent which uses as its primary ingredient phosphoric acid salts of zirconium or titanium is presented. Production methods are discussed and several examples are detailed. Measurements of separating characteristics of some gases using the salts are given.

  8. The development of a weak anion micro-capillary film for protein chromatography.

    PubMed

    Kouyoumdjian, A J M; Lazar, R A; Slater, N K H

    2016-10-14

    In this study, the surface of a microporous walled micro-capillary film (MMCF) was modified into a weak anion exchanger by coupling cyanuric chloride and 2-diethylaminoethylamine (DEAE) to the ethylene-vinyl alcohol (EVOH) matrix. Fourier transform infrared spectroscopy (FTIR) measurements of modified and unmodified MMCFs confirmed the addition of a triazine ring and DEAE onto the membrane. Binding of bovine serum albumin (BSA) at pH 7.2 was found to follow a Langmuir isotherm with a maximum equilibrium binding of 12.4mg BSA/mL adsorbent and 8.2mg BSA/mL adsorbent under static and flow conditions, respectively. The ion exchange capacity, determined by Mohr's titration of chlorine atoms displaced from the functionalised surface, was found to be 195±21μmol Cl(-)/mL of adsorber, comparable to commercial ion exchangers. BSA adsorption onto the ion exchanger was strongly pH-dependant, with an observed reduction in binding above pH 8.2. Frontal experiments of a BSA (5mg/mL) and lysozyme (5mg/mL) mixture demonstrated successful separation of BSA from lysozyme at more than 97% purity as verified by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Separation between similarly charged anionic molecules was also achieved using BSA (5mg/mL) and herring sperm DNA (0.25mg/mL). BSA was extracted at 100% purity, demonstrating the ability of MMCF-DEAE to remove significant DNA contamination from a protein solution. These experiments highlight the potential for MMCFs to be used for fast protein purification in preparative chromatography application.

  9. Elastic properties of protein functionalized nanoporous polymer films

    DOE PAGES

    Charles T. Black; Wang, Haoyu; Akcora, Pinar

    2015-12-16

    Retaining the conformational structure and bioactivity of immobilized proteins is important for biosensor designs and drug delivery systems. Confined environments often lead to changes in conformation and functions of proteins. In this study, lysozyme is chemically tethered into nanopores of polystyrene thin films, and submicron pores in poly(methyl methacrylate) films are functionalized with streptavidin. Nanoindentation experiments show that stiffness of streptavidin increases with decreasing submicron pore sizes. Lysozymes in polystyrene nanopores are found to behave stiffer than the submicron pore sizes and still retain their specific bioactivity relative to the proteins on flat surfaces. Lastly, our results show that proteinmore » functionalized ordered nanoporous polystyrene/poly(methyl methacrylate) films present heterogeneous elasticity and can be used to study interactions between free proteins and designed surfaces.« less

  10. Electronic and molecular properties of an adsorbed protein monolayer probed by two-color sum-frequency generation spectroscopy.

    PubMed

    Dreesen, L; Humbert, C; Sartenaer, Y; Caudano, Y; Volcke, C; Mani, A A; Peremans, A; Thiry, P A; Hanique, S; Frère, J-M

    2004-08-17

    Two-color sum-frequency generation spectroscopy (2C-SFG) is used to probe the molecular and electronic properties of an adsorbed layer of the green fluorescent protein mutant 2 (GFPmut2) on a platinum (111) substrate. First, the spectroscopic measurements, performed under different polarization combinations, and atomic force microscopy (AFM) show that the GFPmut2 proteins form a fairly ordered monolayer on the platinum surface. Next, the nonlinear spectroscopic data provide evidence of particular coupling phenomena between the GFPmut2 vibrational and electronic properties. This is revealed by the occurrence of two doubly resonant sum-frequency generation processes for molecules having both their Raman and infrared transition moments in a direction perpendicular to the sample plane. Finally, our 2C-SFG analysis reveals two electronic transitions corresponding to the absorption and fluorescence energy levels which are related to two different GFPmut2 conformations: the B (anionic) and I forms, respectively. Their observation and wavelength positions attest the keeping of the GFPmut2 electronic properties upon adsorption on the metallic surface.

  11. A synthetic Protein G adsorbent based on the multi-component Ugi reaction for the purification of mammalian immunoglobulins.

    PubMed

    Qian, Jianing; El Khoury, Graziella; Issa, Hamzah; Al-Qaoud, Khaled; Shihab, Penelope; Lowe, Christopher R

    2012-06-01

    Numerous efforts have been devoted to develop synthetic affinity ligands mimicking natural immunoglobulin-binding proteins, such as Proteins A and L, in order to overcome intrinsic drawbacks involving their high cost and acidic pH elution. However, few reports have focused on a Protein G mimic. This work describes the use of the solid phase multi-component Ugi reaction to generate a low cost, rationally designed, affinity ligand to mimic Protein G for the purification of mammalian immunoglobulins, including the heavy-chain only camelid IgGs, with effective elution at neutral pH. An aldehyde-functionalised Sepharose™ resin constituted one component (aldehyde) of the four-component Ugi reaction, whilst the other three components (a primary or secondary amine, a carboxylic acid and an isonitrile) were varied to generate a tri-substituted Ugi scaffold, with a wide range of functionality, suitable for mimicking peptides for immunoglobulin purification. Ligand A2C11I1 was designed to mimic Asn35 and Trp43 of Protein G (PDB: 1FCC) and in silico docking into the Fc domain showed a key binding interface closely resembling native Protein G. This candidate ligand demonstrated affinity towards IgGs derived from human, cow, goat, mouse, sheep, pig, rabbit and rat serum, chicken IgY and recombinant camelid Fc domain, out of which cow and sheep IgG demonstrated 100% binding under the conditions selected. Preparative chromatography of IgG from human serum under a standardised buffer regime eluted IgG of ∼65% purity, compared to ∼62% with Protein G. This adsorbent achieved highest elution of IgG at neutral pH (0.1M sodium phosphate pH 7.0, 30%, v/v, ethylene glycol), an advantage for purifying antibodies sensitive to extremes of pH. The ligand demonstrated a static binding capacity of 24.6 mg Ig G ml⁻¹ resin and a dissociation constant (K(d)) of 4.78 × 10⁻⁶ M. The solid phase Ugi scaffold provides a strategy to develop pseudo-biospecific ligands to purify

  12. New diffusive gradients in a thin film technique for measuring inorganic arsenic and selenium(IV) using a titanium dioxide based adsorbent.

    PubMed

    Bennett, William W; Teasdale, Peter R; Panther, Jared G; Welsh, David T; Jolley, Dianne F

    2010-09-01

    A new diffusive gradients in a thin film (DGT) technique, using a titanium dioxide based adsorbent (Metsorb), has been developed and evaluated for the determination of dissolved inorganic arsenic and selenium. As(III), As(V), and Se(IV) were found to be quantitatively accumulated by the adsorbent (uptake efficiencies of 96.5-100%) and eluted in 1 M NaOH (elution efficiencies of 81.2%, 75.2%, and 88.7%). Se(VI) was not quantitatively accumulated by the adsorbent (<20%). Laboratory DGT validation experiments gave linear mass uptake over time (R(2) >or= 0.998) for As(III), As(V), and Se(IV). Consistent uptake occurred over pH (3.5-8.5) and ionic strength (0.0001-0.75 mol L(-1) NaNO(3)) ranges typical of natural waters, including seawater. Field deployments of DGT probes with various diffusive layer thicknesses confirmed the use of the technique in situ, allowing calculation of the diffusive boundary layers and an accurate measurement of inorganic arsenic. Reproducibility of the technique in field deployments was good (relative standard deviation <8%). Limits of detection (4 day deployments) were 0.01 microg L(-1) for inorganic arsenic and 0.05 microg L(-1) for Se(IV). The results of this study confirmed that DGT with Metsorb was a reliable and robust method for the measurement of inorganic arsenic and the selective measurement of Se(IV) within useful limits of accuracy.

  13. Protein adsorption on dopamine-melanin films: role of electrostatic interactions inferred from zeta-potential measurements versus chemisorption.

    PubMed

    Bernsmann, Falk; Frisch, Benoît; Ringwald, Christian; Ball, Vincent

    2010-04-01

    We recently showed the possibility to build dopamine-melanin films of controlled thickness by successive immersions of a substrate in alkaline solutions of dopamine [F. Bernsmann, A. Ponche, C. Ringwald, J. Hemmerlé, J. Raya, B. Bechinger, J.-C. Voegel, P. Schaaf, V. Ball, J. Phys. Chem. C 113 (2009) 8234-8242]. In this work the structure and properties of such films are further explored. The zeta-potential of dopamine-melanin films is measured as a function of the total immersion time to build the film. It appears that the film bears a constant zeta-potential of (-39+/-3) mV after 12 immersion steps. These data are used to calculate the surface density of charged groups of the dopamine-melanin films at pH 8.5 that are mostly catechol or quinone imine chemical groups. Furthermore the zeta-potential is used to explain the adsorption of three model proteins (lysozyme, myoglobin, alpha-lactalbumin), which is monitored by quartz crystal microbalance. We come to the conclusion that protein adsorption on dopamine-melanin is not only determined by possible covalent binding between amino groups of the proteins and catechol groups of dopamine-melanin but that electrostatic interactions contribute to protein binding. Part of the adsorbed proteins can be desorbed by sodium dodecylsulfate solutions at the critical micellar concentration. The fraction of weakly bound proteins decreases with their isoelectric point. Additionally the number of available sites for covalent binding of amino groups on melanin grains is quantified.

  14. Ultrathin calcium silicate hydrate nanosheets with large specific surface areas: synthesis, crystallization, layered self-assembly and applications as excellent adsorbents for drug, protein, and metal ions.

    PubMed

    Wu, Jin; Zhu, Ying-Jie; Chen, Feng

    2013-09-09

    A simple and low-cost solution synthesis is reported for low-crystalline 1.4 nm tobermorite-like calcium silicate hydrate (CSH) ultrathin nanosheets with a thickness of ~2.8 nm and with a large specific surface area (SSA), via a reaction-rate-controlled precipitation process. The BET SSA of the CSH ultrathin nanosheets can reach as high as 505 m(2) g(-1) . The CSH ultrathin nanosheets have little cytotoxicity and can be converted to anhydrous calcium silicate (ACS) ultrathin nanosheets with a well preserved morphology via a heat treatment process. The crystallinity of CSH ultrathin nanosheets can be improved by solvothermal treatment in water/ethanol binary solvents or a single solvent of water, producing well-crystalline 1.1 nm tobermorite-like CSH nanobelts or nanosheets. CSH ultrathin nanosheets acting as building blocks can self-assemble into layered nanostructures via three different routes. The CSH ultrathin nanosheets are investigated as promising adsorbents for protein (hemoglobin, Hb), drug (ibuprofen, IBU), and metal ions (Cr(3+) , Ni(2+) , Cu(2+) , Zn(2+) , Cd(2+) , Pb(2+) ). The highest adsorbed percentages of Hb and IBU are found to be 83% and 94%, respectively. The highest adsorption capacities of Hb and IBU are found to be as high as 878 milligram Hb per gram CSH and 2.2 gram IBU per gram CSH, respectively. The ppm level metal ions can be totally adsorbed from aqueous solution in just a few minutes. Thus, the CSH ultrathin nanosheets are a promising candidate as excellent adsorbents in the biomedical field and for waste water treatment. Several empirical laws are summarized based on the adsorption profiles of Hb and IBU using CSH ultrathin nanosheets as the adsorbent. Furthermore, the ACS ultrathin nanosheets as adsorbents for Hb protein and IBU drug are investigated.

  15. Nanoscale confinement and interfacial effects on the dynamics and glass transition/crystallinity of thin adsorbed films on silica nanoparticles

    NASA Astrophysics Data System (ADS)

    Madathingal, Rajesh Raman

    The research investigated in this dissertation has focused on understanding the structure-property-function relationships of polymer nanocomposites. The properties of composite systems are dictated by the properties of their components, typically fillers in a polymer matrix. In nanocomposites, the polymer near an interface has significantly different properties compared with the bulk polymer, and the contribution of the adsorbed polymer to composite properties becomes increasingly important as the filler size decreases. Despite many reports of highly favorable properties, the behavior of polymer nanocomposites is not generally predictable, and thus requires a better understanding of the interfacial region. The ability to tailor the filler/matrix interaction and an understanding of the impact of the interface on macroscopic properties are keys in the design of nanocomposite properties. In this original work the surface of silica nanoparticles was tailored by: (a) Changing the number of sites for polymer attachment by varying the surface silanols and, (b) By varying the size/curvature of nanoparticles. The effect of surface tailoring on the dynamic properties after the adsorption of two model polymers, amorphous polymethyl methacrylate (PMMA) and semicrystalline polyethylene oxide (PEO) was observed. The interphase layer of polymers adsorbed to silica surfaces is affected by the surface silanol density as well as the relative size of the polymer compared with the size of the adsorbing substrate. The non-equilibrium adsorption of PMMA onto individual colloidal Stober silica (SiO2) particles, where Rparticle (100nm) > RPMMA (˜6.5nm) was compared with the adsorption onto fumed silica, where Rparticle (7nm) ˜ RPMMA (6.5nm) < Raggregate (˜1000nm), both as a function of silanol density [SiOH] and hydrophobility. In the former case, TEM images showed that the PMMA adsorbed onto individual nanoparticles, so that the number of PMMA chains/bead could be calculated, whereas

  16. Formation of Organized Protein Thin Films with External Electric Field.

    PubMed

    Ferreira, Cecília Fabiana da G; Camargo, Paulo C; Benelli, Elaine M

    2015-10-01

    The effect of an external electric field on the formation of protein GlnB-Hs films and on its buffer solution on siliconized glass slides has been analyzed by current versus electric field curves and atomic force microscopy (AFM). The Herbaspirillum seropedicae GlnB protein (GlnB-Hs) is a globular, soluble homotrimer (36 kDa) with its 3-D structure previously determined. Concentrations of 10 nM native denatured GlnB-Hs protein were deposited on siliconized glass slides under ambient conditions. Immediately after solution deposition a maximum electric field of 30 kV/m was applied with rates of 3 V/s. The measured currents were surface currents and were analyzed as transport current. Electric current started to flow only after a minimum electric field (critical value) for the systems analyzed. The AFM images showed films with a high degree of directional organization only when the proteins were present in the solution. These results showed that the applied electric field favored directional organization of the protein GlnB-Hs films and may contribute to understand the formation of protein films under applied electric fields.

  17. Interaction of Human Plasma Proteins with Thin Gelatin-Based Hydrogel Films: A QCM-D and ToF-SIMS Study

    PubMed Central

    2015-01-01

    In the fields of surgery and regenerative medicine, it is crucial to understand the interactions of proteins with the biomaterials used as implants. Protein adsorption directly influences cell-material interactions in vivo and, as a result, regulates, for example, cell adhesion on the surface of the implant. Therefore, the development of suitable analytical techniques together with well-defined model systems allowing for the detection, characterization, and quantification of protein adsorbates is essential. In this study, a protocol for the deposition of highly stable, thin gelatin-based films on various substrates has been developed. The hydrogel films were characterized morphologically and chemically. Due to the obtained low thickness of the hydrogel layer, this setup allowed for a quantitative study on the interaction of human proteins (albumin and fibrinogen) with the hydrogel by Quartz Crystal Microbalance with Dissipation Monitoring (QCM-D). This technique enables the determination of adsorbant mass and changes in the shear modulus of the hydrogel layer upon adsorption of human proteins. Furthermore, Secondary Ion Mass Spectrometry and principal component analysis was applied to monitor the changed composition of the topmost adsorbate layer. This approach opens interesting perspectives for a sensitive screening of viscoelastic biomaterials that could be used for regenerative medicine. PMID:24956040

  18. dc-Sheet resistance as sensitive monitoring tool of protein immobilization on thin metal films.

    PubMed

    Neff, H; Beeby, T; Lima, A M N; Borre, M; Thirstrup, C; Zong, W; de Almeida, L A L

    2006-03-15

    The suitability of high resolution, in situ dc-sheet resistance monitoring (SRM) as a simplified and reliable sensing technique towards detection and tracking of protein immobilization has been explored. Non-specific adsorption of bovine serum albumin (BSA) onto a very thin gold film, acting as the sensing resistor, has been employed as a model system. For comparison, the novel sensing method was combined with surface plasmon resonance (SPR) spectroscopy, using the same flow cell and sensing surface. Two different, well known adsorption states, involving a composite layer of irreversibly and reversibly bound BSA, were clearly resolved by both methods. Clearly structured, pronounced and fully reproducible film resistance modulations have been resolved in the associated SRM data. The transition from reversibly bound BSA to the diluted protein phase is associated with an unusually large decrease in the dc-sheet resistance. The observed resistance modulation magnitude for an adsorbed BSA monolayer corresponds to approximately 1%, and up to 100 mOmega at a 10 Omega sensing resistor. The sheet resistance of irreversibly bound BSA was determined to 0.24 kOmega/cm2, and the associated specific resistivity estimated to 1-2x10(4) Omega cm.

  19. Quantifying protein adsorption on combinatorially sputtered Al-, Nb-, Ta- and Ti-containing films with electron microprobe and spectroscopic ellipsometry

    NASA Astrophysics Data System (ADS)

    Byrne, T. M.; Lohstreter, L.; Filiaggi, M. J.; Bai, Zhijun; Dahn, J. R.

    2009-04-01

    Although metallic biomaterials are widely used, systematic studies of protein adsorption onto such materials are generally lacking. Combinatorial binary libraries of Al 1-xNb x, Al 1-xTa x, Al 1-xTi x, Nb 1-xTa x, Nb 1-xTi x, and Ta 1-xTi x (0 ⩽ x ⩽ 1) and a ternary library of Al 1-xTi xTa y (0 ⩽ x ⩽ 1 and 0 ⩽ y ⩽ 0.7), along with their corresponding pure element films were sputtered onto glass substrates using a unique magnetron sputtering technique. Films were characterized with wavelength-dispersive spectroscopy (WDS), X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS). Fibrinogen and albumin adsorption amounts were measured by wavelength-dispersive spectroscopy (WDS) and spectroscopic ellipsometry (SE) equipment, both high throughput techniques with automated motion stage capabilities. Protein adsorption onto these films was found to be closely correlated to the alumina surface fraction, with high alumina content at the surface leading to low amounts of adsorbed fibrinogen and albumin. Protein adsorption amounts obtained with WDS and SE were in good agreement for all films.

  20. Multilayered films fabricated from an oligoarginine-conjugated protein promote efficient surface-mediated protein transduction.

    PubMed

    Jewell, Christopher M; Fuchs, Stephen M; Flessner, Ryan M; Raines, Ronald T; Lynn, David M

    2007-03-01

    The conjugation of cationic protein transduction domains to proteins results in an increase in the extent to which proteins are internalized by cells. This investigation sought to determine whether the conjugation of a protein transduction domain to a functional protein could be used to facilitate the incorporation of the protein into multilayered polyelectrolyte films and, subsequently, whether these films could be used to promote surface-mediated protein transduction. We demonstrate that it is possible to fabricate multilayered assemblies 80 nm thick using sodium polystyrene sulfonate (SPS) and bovine pancreatic ribonuclease (RNase A) conjugated to the cationic protein transduction domain nonaarginine (R(9)) using an entirely aqueous layer-by-layer process. We demonstrate further that the conjugation of R(9) to RNase A permits the assembly of multilayered films under conditions that do not allow for the incorporation of the unmodified protein. This result suggests that R(9) functions as a cationic anchor and serves to increase the strength of electrostatic interactions with SPS and facilitate layer-by-layer assembly. We also demonstrate that RNase A-R(9)/SPS films dissolve rapidly in physiologically relevant media and that macroscopic objects coated with these materials can be used to mediate high levels of protein transduction in mammalian cells. These results suggest the basis of general methods that could contribute to the design of materials that permit spatial and temporal control over the delivery of therapeutic proteins to cells and tissues.

  1. Direct observation of intraparticle equilibration and the rate-limiting step in adsorption of proteins in chromatographic adsorbents with confocal laser scanning microscopy.

    PubMed

    Kasche, Volker; de Boer, Michael; Lazo, Cesar; Gad, Moustafa

    2003-06-25

    The adsorption of different proteins in a single biospecific and hydrophobic adsorbent particle for preparative protein chromatography has been observed directly by confocal laser scanning microscopy as a function of time at a constant bulk concentration c(b). The bulk concentration was in the non-linear part of the adsorption isotherm. At all times the concentration of free protein at the particle surface was almost equal to the bulk content indicating that external mass transfer resistance is not rate limiting for the adsorption under these conditions. Inside the particles a distinct maximum in adsorbed and free protein concentration that moved inside to a distance of approximately 0.2 R (R particle radius) from the particle surface, was observed. This is due to a decreasing solid-phase density and adsorptive capacity in the particle between 0.8 R and R indicating that the fraction of macropores (or void space) is larger in the outer than in the inner part of the adsorbent particles. By increasing the bulk concentration by a factor of 10 the equilibration time was reduced by about the same magnitude. This is in agreement with the concentration dependence of the effective pore diffusion coefficient D(p,eff)=D(p)/[epsilon(p)[1+nK/(K +c)(2)

  2. Charge transport in disordered films of non-redox proteins

    NASA Astrophysics Data System (ADS)

    Pompa, P. P.; Della Torre, A.; del Mercato, L. L.; Chiuri, R.; Bramanti, A.; Calabi, F.; Maruccio, G.; Cingolani, R.; Rinaldi, R.

    2006-07-01

    Electrical conduction in solid state disordered multilayers of non-redox proteins is demonstrated by two-terminal transport experiments at the nanoscale and by scanning tunneling microscopy (STM/STS experiments). We also show that the conduction of the biomolecular films can be modulated by means of a gate field. These results may lead to the implementation of protein-based three-terminal nanodevices and open important new perspectives for a wide range of bioelectronic/biosensing applications.

  3. Electrospun polystyrene/graphene nanofiber film as a novel adsorbent of thin film microextraction for extraction of aldehydes in human exhaled breath condensates.

    PubMed

    Huang, Jing; Deng, Hongtao; Song, Dandan; Xu, Hui

    2015-06-09

    In the current study, we introduced a novel polystyrene/graphene (PS/G) composite nanofiber film for thin film microextraction (TFME) for the first time. The PS/G nanofiber film was fabricated on the surface of filter paper by a facile electrospinning method. The morphology and extraction performance of the resultant composite film were investigated systematically. The PS/G nanofiber film exhibited porous fibrous structure, large surface area and strong hydrophobicity. A new thin film microextraction-high performance liquid chromatography (TFME-HPLC) method was developed for the determination of six aldehydes in human exhaled breath condensates. The method showed high enrichment efficiency and fast analysis speed. Under the optimal conditions, the linear ranges of the analytes were in the range of 0.02-30 μmol L(-1) with correlation coefficients above 0.9938, and the recoveries were between 79.8% and 105.6% with the relative standard deviation values lower than 16.3% (n=5). The limits of quantification of six aldehydes ranged from 13.8 to 64.6 nmol L(-1). The established method was successfully applied for the quantification of aldehyde metabolites in exhaled breath condensates of lung cancer patients and healthy people. Taken together, the TFME-HPLC method provides a simple, rapid, sensitive, cost-effective, non-invasion approach for the analysis of linear aliphatic aldehydes in human exhaled breath condensates.

  4. Spontaneously adsorbed monolayer films: Fabrication, characterization, and application of monolayers of alkanethiol and sulfur-bearing cyclodestrin derivatives

    SciTech Connect

    Chinkap, Chung.

    1991-03-12

    Monolayers of n-alkanethiols (CH{sub 3}(CH{sub 2}){sub n}SH, n=1--17) and sulfur-bearing cyclodextrin derivatives spontaneously adsorbed on Ag and Au have been studied with a variety of surface characterization methods, such as infrared inflection spectroscopy, contact angle measurements, electro-chemistry, optical ellipsometry, and scanning tunneling microscopy. Long chain n-alkanethiols monolayers on Ag and Au are insulating to electron transfer and have contact angles indicative of well-ordered hydrocarbon terminated structures. Infrared and contact angle data indicate a different orientation of the methyl group with respect to the surface for chains with odd and even numbers of methylene groups. Compared to monolayers on Au, the alkanethiol monolayers on Ag are oriented more towards the surface normal. The observed odd-even effect methyl group orientation for these monolayers on Ag is offset by a methylene group from that on Au. The relationships between the structure and packing of the monolayers on Ag and Au and the composition, roughness, and crystallinity of the substrate are also discussed. Monolayers of sulfur-bearing cyclodextrin derivatives on Au and Ag are fabricated by spontaneous adsorption and characterized by the above techniques. Size-selectively and molecular recognition of the {alpha}- and {beta}- cyclodextrin cavity are shown with our monolayers. Because of molecular recognition, p-nitrophenol is retained preferrentially by the cyclodextrin monolayers over o-nitrophenol. 146 refs., 44 figs., 5 tabs.

  5. Nanostructured thin film polymer devices for constant-rate protein delivery.

    PubMed

    Bernards, Daniel A; Lance, Kevin D; Ciaccio, Natalie A; Desai, Tejal A

    2012-10-10

    Herein long-term delivery of proteins from biodegradable thin film devices is demonstrated, where a nanostructured polymer membrane controls release. Protein was sealed between two poly(caprolactone) films, which generated the thin film devices. Protein release for 210 days was shown in vitro, and stable activity was established through 70 days with a model protein. These thin film devices present a promising delivery platform for biologic therapeutics, particularly for application in constrained spaces.

  6. Protein-modified nanocrystalline diamond thin films for biosensor applications

    NASA Astrophysics Data System (ADS)

    Härtl, Andreas; Schmich, Evelyn; Garrido, Jose A.; Hernando, Jorge; Catharino, Silvia C. R.; Walter, Stefan; Feulner, Peter; Kromka, Alexander; Steinmüller, Doris; Stutzmann, Martin

    2004-10-01

    Diamond exhibits several special properties, for example good biocompatibility and a large electrochemical potential window, that make it particularly suitable for biofunctionalization and biosensing. Here we show that proteins can be attached covalently to nanocrystalline diamond thin films. Moreover, we show that, although the biomolecules are immobilized at the surface, they are still fully functional and active. Hydrogen-terminated nanocrystalline diamond films were modified by using a photochemical process to generate a surface layer of amino groups, to which proteins were covalently attached. We used green fluorescent protein to reveal the successful coupling directly. After functionalization of nanocrystalline diamond electrodes with the enzyme catalase, a direct electron transfer between the enzyme's redox centre and the diamond electrode was detected. Moreover, the modified electrode was found to be sensitive to hydrogen peroxide. Because of its dual role as a substrate for biofunctionalization and as an electrode, nanocrystalline diamond is a very promising candidate for future biosensor applications.

  7. A biomimetic Protein G affinity adsorbent: an Ugi ligand for immunoglobulins and Fab fragments based on the third IgG-binding domain of Protein G.

    PubMed

    El Khoury, Graziella; Lowe, Christopher R

    2013-04-01

    This work reports the development of a synthetic affinity adsorbent for immunoglobulins based on the Fab-binding domain of Streptococcal Protein G (SpG-domain III). The ligand (A2C7I1) was synthesized by the four-component Ugi reaction to generate a substituted peptoidal scaffold mimicking key amino acid residues of SpG. Computer-aided analysis suggests a putative binding site on the CH 1 domain of the Fab molecule. In silico studies, supported by affinity chromatography in comparison with immobilized SpG, as well as analytical characterization by liquid chromatography/electrospray ionization-mass spectrometry and (1) H nuclear magnetic resonance of the ligand synthesized in solution, indicated the authenticity and suitability of the designed ligand for the purification of immunoglobulins. The immobilized ligand displayed an apparent static binding capacity of ~17 mg IgG ml(-1) and a dissociation constant of 5.34 × 10(-5)  M. Preparative chromatography demonstrated the ability of the immobilized ligand to purify IgG and Fab fragments from crude mammalian and yeast cell cultures, under near physiological ionic strength and pH, to yield proteins of 99% and 93% purity, respectively.

  8. Surface-enhanced Raman scattering of 4-aminothiophenol adsorbed on silver nanosheets deposited onto cubic boron nitride films.

    PubMed

    Zhou, Yanli; Zhi, Jinfang; Zhao, Jianwen; Xu, Maotian

    2010-01-01

    A simple method was found for the fabrication of silver nanosheets (AgNS) by the catalysis of gold nanoparticles (AuNP) on an amine-terminated cubic boron nitride (cBN) surface deposited on a Si(001) substrate in the presence of reductant. The morphology of the AgNS/AuNP/NH(2)-cBN/Si(001) sample was characterized by scanning electron microscopy and X-ray diffraction. The performance of the AgNS/AuNP/NH(2)-cBN/Si(001) sample as surface-enhanced Raman scattering (SERS) active substrate was evaluated by using 4-aminothiophenol (PATP) as the probe molecule. The SERS measurements showed that the maximum intensity was obtained on the AgNS/AuNP/NH(2)-cBN/Si(001) sample for 5 min silver deposition. Compared with the AuNP/NH(2)-cBN/Si(001) sample and a silver film/cBN/Si(001) prepared by the mirror reaction, the SERS signal of PATP was obviously improved on the above AgNS/AuNP/NH(2)-cBN/Si(001) film. The sensitivity and the stability of the AgNS/AuNP/NH(2)-cBN/Si(001) sample were also investigated.

  9. Lotus-leaf-like topography predominates over adsorbed ECM proteins in poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) surface/cell interactions.

    PubMed

    Zheng, Jun; Li, Dan; Yuan, Lin; Liu, Xiaoli; Chen, Hong

    2013-06-26

    It is well-known that extracellular matrix (ECM) proteins mediate cell/surface interactions. However, introduction of a specific surface topography may disturb the correlation between ECM proteins adsorption and cells adhesion on a given surface. In present study, lotus-leaf-like topography was introduced on the surface of a biodegradable material, poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx). Protein adsorption and cell interactions with this lotus-leaf-like surface (designated PHBHHx-L) were investigated. Water contact angle data indicated that the hydrophobicity of PHBHHx was enhanced by the introduction of lotus-leaf-like topography. The adsorption of extracellular matrix proteins (fibronectin and vitronectin) on PHBHHx-L was measured by enzyme linked immunosorbent assay (ELISA). Compared with flat PHBHHx, adsorption on the PHBHHx-L surface increased by ~260% for fibronectin and ~40% for vitronectin. In contrast, fibroblast and endothelial cell adhesion and proliferation were reduced on the PHBHHx-L compared to the flat polymer surface. These results suggest that the inhibition of cell adhesion and proliferation caused by the lotus-leaf-like topography dominates over the effect of the adsorbed adhesive proteins in promoting adhesion and proliferation. It can be concluded that the lotus-leaf-like topography plays a dominant role in cell/PHBHHx-L interactions. The present findings indicate the complexity of the interplay among surface topography, adsorbed proteins, and cell-surface interactions.

  10. Monomolecular films of cholesterol oxidase and S-Layer proteins

    NASA Astrophysics Data System (ADS)

    Ferraz, Helen Conceição; Guimarães, Juliana Aguilar; Alves, Tito Livio Moitinho; Constantino, Carlos José Leopoldo

    2011-05-01

    Cholesterol oxidase (ChOx) is a flavoenzyme that catalyzes the oxidation of cholesterol to cholest-5-en-3-one and subsequently the isomerization to cholest-4-en-3-one. ChOx has been very commonly studied as the detection element in cholesterol biosensors. In the biosensor development field, a relatively new approach is the use of crystalline bacterial cell surface layers, known as S-Layer proteins. These proteins exhibit the ability of self-assembling at surfaces, opening a vast spectrum of applications, both in basic and applied researches. In our study, monomolecular films of ChOx and mixed films of ChOx/S-Layer proteins and DPPC/S-Layer proteins were produced using the Langmuir technique. Characterization of the films was performed by means of surface pressure-molecular area ( π- A) isotherms. Stable monolayers were obtained, which means that they can be transferred to solid substrates by Langmuir-Blodgett technique. Mixed monolayers showed an ideal like behavior.

  11. Driving force behind adsorption-induced protein unfolding: a time-resolved X-ray reflectivity study on lysozyme adsorbed at an air/water interface.

    PubMed

    Yano, Yohko F; Uruga, Tomoya; Tanida, Hajime; Toyokawa, Hidenori; Terada, Yasuko; Takagaki, Masafumi; Yamada, Hironari

    2009-01-06

    Time-resolved X-ray reflectivity measurements for lysozyme (LSZ) adsorbed at an air/water interface were performed to study the mechanism of adsorption-induced protein unfolding. The time dependence of the density profile at the air/water interface revealed that the molecular conformation changed significantly during adsorption. Taking into account previous work using Fourier transform infrared (FTIR) spectroscopy, we propose that the LSZ molecules initially adsorbed on the air/water interface have a flat unfolded structure, forming antiparallel beta-sheets as a result of hydrophobic interactions with the gas phase. In contrast, as adsorption continues, a second layer forms in which the molecules have a very loose structure having random coils as a result of hydrophilic interactions with the hydrophilic groups that protrude from the first layer.

  12. Making novel bio-interfaces through bacterial protein recrystallization on biocompatible polylactide derivative films

    NASA Astrophysics Data System (ADS)

    Lejardi, Ainhoa; López, Aitziber Eleta; Sarasua, José R.; Sleytr, U. B.; Toca-Herrera, José L.

    2013-09-01

    Fabrication of novel bio-supramolecular structures was achieved by recrystallizing the bacterial surface protein SbpA on amorphous and semicrystalline polylactide derivatives. Differential scanning calorimetry showed that the glass transition temperature (Tg) for (poly-L-lactide)-PLLA, poly(L,D-lactide)-PDLLA, poly(lactide-co-glycolide)-PLGA and poly(lactide-co-caprolactone)-PLCL was 63 °C, 53 °C, 49 °C and 15 °C, respectively. Tensile stress-strain tests indicated that PLLA, PLGA, and PDLLA had a glassy behaviour when tested below Tg. The obtained Young modulus were 1477 MPa, 1330 MPa, 1306 MPa, and 9.55 MPa for PLLA, PLGA, PDLLA, and PLCL, respectively. Atomic force microscopy results confirmed that SbpA recrystallized on every polymer substrate exhibiting the native S-layer P4 lattice (a = b = 13 nm, γ = 90°). However, the polymer substrate influenced the domain size of the S-protein crystal, with the smallest size for PLLA (0.011 μm2), followed by PDLLA (0.034 μm2), and PLGA (0.039 μm2), and the largest size for PLCL (0.09 μm2). quartz crystal microbalance with dissipation monitoring (QCM-D) measurements indicated that the adsorbed protein mass per unit area (˜1800 ng cm-2) was independent of the mechanical, thermal, and crystalline properties of the polymer support. The slowest protein adsorption rate was observed for amorphous PLCL (the polymer with the weakest mechanical properties and lowest Tg). QCM-D also monitored protein self-assembly in solution and confirmed that S-layer formation takes place in three main steps: adsorption, self-assembly, and crystal reorganization. Finally, this work shows that biodegradable polylactide derivatives films are a suitable support to form robust biomimetic S-protein layers.

  13. Integrated system for temperature-controlled fast protein liquid chromatography comprising improved copolymer modified beaded agarose adsorbents and a travelling cooling zone reactor arrangement.

    PubMed

    Müller, Tobias K H; Cao, Ping; Ewert, Stephanie; Wohlgemuth, Jonas; Liu, Haiyang; Willett, Thomas C; Theodosiou, Eirini; Thomas, Owen R T; Franzreb, Matthias

    2013-04-12

    An integrated approach to temperature-controlled chromatography, involving copolymer modified agarose adsorbents and a novel travelling cooling zone reactor (TCZR) arrangement, is described. Sepharose CL6B was transformed into a thermoresponsive cation exchange adsorbent (thermoCEX) in four synthetic steps: (i) epichlorohydrin activation; (ii) amine capping; (iii) 4,4'-azobis(4-cyanovaleric acid) immobilization; and 'graft from' polymerization of poly(N-isopropylacrylamide-co-N-tert-butylacrylamide-co-acrylic acid-co-N,N'-methylenebisacrylamide). FT-IR, (1)H NMR, gravimetry and chemical assays allowed precise determination of the adsorbent's copolymer composition and loading, and identified the initial epoxy activation step as a critical determinant of 'on-support' copolymer loading, and in turn, protein binding performance. In batch binding studies with lactoferrin, thermoCEX's binding affinity and maximum adsorption capacity rose smoothly with temperature increase from 20 to 50 °C. In temperature shifting chromatography experiments employing thermoCEX in thermally jacketed columns, 44-51% of the lactoferrin adsorbed at 42 °C could be desorbed under binding conditions by cooling the column to 22 °C, but the elution peaks exhibited strong tailing. To more fully exploit the potential of thermoresponsive chromatography adsorbents, a new column arrangement, the TCZR, was developed. In TCZR chromatography, a narrow discrete cooling zone (special assembly of copper blocks and Peltier elements) is moved along a bespoke fixed-bed separation columnfilled with stationary phase. In tests with thermoCEX, it was possible to recover 65% of the lactoferrin bound at 35 °C using 8 successive movements of the cooling zone at a velocity of 0.1mm/s; over half of the recovered protein was eluted in the first peak in more concentrated form than in the feed. Intra-particle diffusion of desorbed protein out of the support pores, and the ratio between the velocities of the cooling

  14. Suspended crystalline films of protein hydrophobin I (HFBI).

    PubMed

    Oude Vrielink, Anneloes S; Bomans, Paul H H; Vredenbregt, Edgar J D; Wirix, Maarten J M; Sommerdijk, Nico A J M; Luiten, O J; Voets, Ilja K

    2015-06-01

    Protein interfaces play an essential role in both natural and man-made materials as stabilizers, sensors, catalysts, and selective channels for ions and small molecules. Probing the molecular arrangement within such interfaces is of prime importance to understand the relation between structure and functionality. Here we report on the preparation and characterization of large area suspended crystalline films of class II hydrophobin HFBI. This small, amphiphilic globular protein readily self-assembles at the air-water interface into a 2D hexagonal lattice which can be transferred onto a holey carbon electron microscopy grid yielding large areas of hundreds of square micrometers intact hydrophobin film spun across micron-sized holes. Fourier transform analysis of low-dose electron microscopy images and selected area electron diffraction profiles reveal a unit cell dimension a=5.6±0.1nm, in agreement with reported atomic force microscopy studies on solid substrates and grazing incidence X-ray scattering experiments at the air-water interface. These findings constitute the first step towards the utilization of large-area suspended crystalline hydrophobin films as membranes for ultrapurification and chiral separation or as biological substrates for ultrafast electron diffraction.

  15. Impact of temperature and electrical potentials on the stability and structure of collagen adsorbed on the gold electrode

    NASA Astrophysics Data System (ADS)

    Meiners, Frank; Ahlers, Michael; Brand, Izabella; Wittstock, Gunther

    2015-01-01

    The morphology and structure of collagen type I adsorbed on gold electrodes were studied as a function of electrode potential and temperature by means of capacitance measurements, polarization modulation infrared reflection-absorption spectroscopy and scanning force microscopy at temperatures of 37 °C, 43 °C and 50 °C. The selected temperatures corresponded to the normal body temperature, temperature of denaturation of collagen molecules and denaturation of collagen fibrils, respectively. Independently of the solution temperature, collagen was adsorbed on gold electrodes in the potential range - 0.7 V < E < 0.4 V vs. Ag/AgCl, where the protein film was very stable. Fragments of collagen molecules made a direct contact to the gold surface and water was present in the film. Protein molecules were oriented preferentially with their long axis towards the gold surface. Collagen molecules in the adsorbed state preserved their native triple helical structure even at temperatures corresponding to collagen denaturation in aqueous solutions. Application of E < - 0.75 V vs. Ag/AgCl leads to the swelling of the protein film by water and desorption from the electrode surface. IR spectra provided no evidence of the thermal denaturation of adsorbed collagen molecules. A temperature increase to 50 °C leads to a distortion of the collagen film. The processes of aggregation and fibrilization were preferred over thermal denaturation for collagen adsorbed on the electrode surface and exposed to changing potentials.

  16. Xanthan hydrogel films: molecular conformation, charge density and protein carriers.

    PubMed

    Bueno, Vânia Blasques; Petri, Denise Freitas Siqueira

    2014-01-30

    In this article the molecular conformation of xanthan chains in hydrogel films was investigated by means of circular dichroism, showing substantial differences between xanthan hydrogel prepared in the absence (XNT) and in the presence of citric acid (XCA). The xanthan chains in XNT hydrogels films presented ordered conformation (helixes), while in XCA they were in the disordered conformation (coils), exposing a larger number of carboxylate groups than XNT. The large charge density in XCA hydrogels was evidenced by their behavior under variable ionic strength. Studies about the application of XNT and XCA for loading and delivering of bovine serum albumin (BSA) and lysozyme (LYZ) showed that both events are controlled by hydrogels and proteins net charge, which can be triggered by pH. The preservation of LYZ native conformation after hydrogel loading explained the substantial bactericidal activity of LYZ loaded hydrogels and enables their use as active wound dressings.

  17. Protein patterning on silicon-based surface using background hydrophobic thin film.

    PubMed

    Lee, Chang-Soo; Lee, Sang-Ho; Park, Sung-Soo; Kim, Yong-Kweon; Kim, Byung-Gee

    2003-04-01

    A new and convenient protein patterning method on silicon-based surface was developed for protein array by spin coating of hydrophobic thin film (CYTOP). Photolithographic lift-off process was used to display two-dimensional patterns of spatially hydrophilic region. The background hydrophobic thin film was used to suppress nonspecific protein binding, and the hydrophilic target protein binding region was chemically modified to introduce aldehyde group after removal of the photoresist layer. The difference in surface energy between the hydrophilic pattern and background hydrophobic film would induce easier covalent binding of proteins onto defined hydrophilic areas having physical and chemical constraints. Below 1 microg/ml of total protein concentration, the CYTOP hydrophobic film effectively suppressed nonspecific binding of the protein. During the process of protein patterning, inherent property of the hydrophobic thin film was not changed judging from static and dynamic contact angle survey. Quantitative analysis of the protein binding was demonstrated by streptavidin-biotin system.

  18. Surface characteristics and protein adsorption on combinatorial binary Ti-M (Cr, Al, Ni) and Al-M (Ta, Zr) library films.

    PubMed

    Bai, Zhijun; Filiaggi, M J; Sanderson, R J; Lohstreter, L B; McArthur, M A; Dahn, J R

    2010-02-01

    Systematic studies of protein adsorption onto metallic biomaterial surfaces are generally lacking. Here, combinatorial binary library films with compositional gradients of Ti(1-x)Cr(x), Ti(1-x)Al(x), Ti(1-x)Ni(x) and Al(1-x)Ta(x), (0 films were sputtered onto clean Si surfaces. Bulk and surface chemistry, film microstructure, and surface roughness were subsequently correlated to fibrinogen or albumin adsorption measured using a high throughput wavelength dispersive spectroscopy technique. X-ray diffraction revealed these binary films to have crystalline phases present primarily at either extreme of the compositional library and an amorphous zone dominating along the gradient. These mirror-like films were generally found by atomic force microscopy to have a roughness of less than 8 nm, with any relative increases in roughness consistent with the development of crystalline phases. Surface chemistry by quantitative high-resolution X-ray photoelectron spectroscopy differed significantly from bulk film composition as measured by electron microprobe, with TiO(2) and Al(2)O(3) preferentially forming on the binary film surfaces. Correspondingly, protein adsorption onto these films closely correlated with their surface oxide fractions. Aluminum deposited as either a constant-composition film or as part of a binary library consistently adsorbed the least amount of albumin and fibrinogen, with alumina-enrichment of the surface oxide correlating with this adsorption. Overall, this combinatorial materials approach coupled with high-throughput surface analytical methods provides an efficient method of screening potential metallic biomaterials that may enable as well systematic studies of surface properties driving protein adsorption on these metal / metal oxide systems.

  19. Development of soybean protein-isolate edible films incorporated with beeswax, Span 20, and glycerol.

    PubMed

    Chao, Zhang; Yue, Ma; Xiaoyan, Zhao; Dan, Ma

    2010-08-01

    The effect of the beeswax, Span 20, and glycerol content on qualities of soybean-protein-isolate edible films was evaluated. Beeswax and Span 20 were selected to improve qualities of soybean-protein-isolate films from 11 emulsifiers. The content of beeswax, Span 20, and glycerol was further optimized by response surface analysis. The optimal composite emulsifier was beeswax (1.87% of soybean protein-isolate), Span 20 (10.25% of soybean protein-isolate), and glycerol (29.12% of soybean protein-isolate) with tensile strength of 908 MPa, percentage elongation at break of 25.8%, water vapor permeability of 19.2 g/m.d.MPa, and oxygen permeability of 0 cm(3)/m.d.MPa. The quality of soybean-protein-isolate films incorporated with the optimal composite emulsifier was 2.34 times higher than that of the control. Furthermore, the disulfide bond content of soybean-protein-isolate films showed a positive correlation with their quality, which provided a simple and rapid way to rank quality of soybean-protein-isolate films. Therefore, our result will not only give an instruction to soybean-protein-isolate-film production, but also give a simple and rapid way to rank film qualities. Practical Application: Our results give the optimal composite emulsifiers for the soybean-protein-isolate-film production. The soybean-protein-isolate films based on the optimal composite emulsifiers show their tensile strength of 908 MPa, percentage elongation at break of 25.8%, water vapor permeability of 19.2 g/m.d.MPa, and oxygen permeability of 0 cm(3)/m.d.MPa, being stronger than the control. Moreover, our results give a simple and rapid way to rank film qualities, because the disulfide bond content of soybean-protein-isolate films showed a positive correlation with their quality. Hence, the disulfide bond content was an indicator to rank qualities of soybean-protein-isolate films.

  20. Differential dehydration effects on globular proteins and intrinsically disordered proteins during film formation.

    PubMed

    Yoneda, Juliana Sakamoto; Miles, Andew J; Araujo, Ana Paula Ulian; Wallace, B A

    2017-04-01

    Globular proteins composed of different secondary structures and fold types were examined by synchrotron radiation circular dichroism spectroscopy to determine the effects of dehydration on their secondary structures. They exhibited only minor changes upon removal of bulk water during film formation, contrary to previously reported studies of proteins dehydrated by lyophilization (where substantial loss of helical structure and gain in sheet structure was detected). This near lack of conformational change observed for globular proteins contrasts with intrinsically disordered proteins (IDPs) dried in the same manner: the IDPs, which have almost completely unordered structures in solution, exhibited increased amounts of regular (mostly helical) secondary structures when dehydrated, suggesting formation of new intra-protein hydrogen bonds replacing solvent-protein hydrogen bonds, in a process which may mimic interactions that occur when IDPs bind to partner molecules. This study has thus shown that the secondary structures of globular and intrinsically disordered proteins behave very differently upon dehydration, and that films are a potentially useful format for examining dehydrated soluble proteins and assessing IDPs structures.

  1. Differential dehydration effects on globular proteins and intrinsically disordered proteins during film formation

    PubMed Central

    Yoneda, Juliana Sakamoto; Miles, Andew J.; Araujo, Ana Paula Ulian

    2017-01-01

    Abstract Globular proteins composed of different secondary structures and fold types were examined by synchrotron radiation circular dichroism spectroscopy to determine the effects of dehydration on their secondary structures. They exhibited only minor changes upon removal of bulk water during film formation, contrary to previously reported studies of proteins dehydrated by lyophilization (where substantial loss of helical structure and gain in sheet structure was detected). This near lack of conformational change observed for globular proteins contrasts with intrinsically disordered proteins (IDPs) dried in the same manner: the IDPs, which have almost completely unordered structures in solution, exhibited increased amounts of regular (mostly helical) secondary structures when dehydrated, suggesting formation of new intra‐protein hydrogen bonds replacing solvent‐protein hydrogen bonds, in a process which may mimic interactions that occur when IDPs bind to partner molecules. This study has thus shown that the secondary structures of globular and intrinsically disordered proteins behave very differently upon dehydration, and that films are a potentially useful format for examining dehydrated soluble proteins and assessing IDPs structures. PMID:28097742

  2. Functional properties and applications of edible films made of milk proteins.

    PubMed

    Chen, H

    1995-11-01

    Edible films and coatings based on milk proteins have been developed to be used as a protective layer on foods or between food components. The most important functionalities of an edible film or coating include control of mass transfers, mechanical protection, and sensory appeal. Control of mass transfers involves preventing foods from desiccation, regulating microenvironments of gases around foods, and controlling migration of ingredients and additives in the food systems. Adequate mechanical strength of an edible film is necessary to protect the integrity of packaging throughout distribution. The sensory properties of an edible coating or film are a key factor for acceptance of final products. Simple milk protein films are good barriers to gas transfers because of their complex intermolecular bindings. Lipid is frequently incorporated into protein films to improve their properties as barriers to moisture vapor. Protein films are distinctly different in mechanical profiles from those films made of other materials. Approaches traditionally used in material sciences have been adapted and modified for studying the functionality of edible films. Potential uses of innovative processing technologies in film making to alter the film functionality are briefly discussed. A survey of potential applications of edible film based on milk protein is presented.

  3. Slide Agglutination Method for the Serological Identification of Neisseria gonorrhoeae with Anti-Gonococcal Antibodies Adsorbed to Protein A-Containing Staphylococci

    PubMed Central

    Danielsson, Dan; Kronvall, Göran

    1974-01-01

    A rapid slide agglutination test has been developed for the identification of Neisseria gonorrhoeae that are primarily detected as oxidase-positive colonies in gonococcal cultures. The technique is based on the specific nonimmune reactivity between the Fc portion of immunoglobulin (Ig)G and staphylococcal protein A. IgG molecules adsorbed to stabilized staphylococci will thereby become oriented with their antigen-reactive sites that are directed outwards. Protein A-containing staphylococci with unabsorbed anti-gonococcal antibodies gave positive co-agglutination reactions with gonococci but also with meningococci, some Moraxella, Haemophilus, and Pseudomonas strains. These crossreactions were eliminated by absorption of the anti-gonococcal antiserum with meningococcal and Moraxella organisms prior to the coating of reagent staphylococci. In the routine culture diagnosis of N. gonorrhoeae the use of specific gonococcal reagent staphylococci gave concordant results with fermentation procedures and immunofluorescent techniques. Images PMID:4207280

  4. Ambient DESI and LESA-MS Analysis of Proteins Adsorbed to a Biomaterial Surface Using In-Situ Surface Tryptic Digestion

    NASA Astrophysics Data System (ADS)

    Rao, Wei; Celiz, Adam D.; Scurr, David J.; Alexander, Morgan R.; Barrett, David A.

    2013-12-01

    The detection and identification of proteins adsorbed onto biomaterial surfaces under ambient conditions has significant experimental advantages but has proven to be difficult to achieve with conventional measuring technologies. In this study, we present an adaptation of desorption electrospray ionization (DESI) and liquid extraction surface analysis (LESA) mass spectrometry (MS) coupled with in-situ surface tryptic digestion to identify protein species from a biomaterial surface. Cytochrome c, myoglobin, and BSA in a combination of single and mixture spots were printed in an array format onto Permanox slides, followed by in-situ surface digestion and detection via MS. Automated tandem MS performed on surface peptides was able to identify the proteins via MASCOT. Limits of detection were determined for DESI-MS and a comparison of DESI and LESA-MS peptide spectra characteristics and sensitivity was made. DESI-MS images of the arrays were produced and analyzed with imaging multivariate analysis to automatically separate peptide peaks for each of the proteins within a mixture into distinct components. This is the first time that DESI and LESA-MS have been used for the in-situ detection of surface digested proteins on biomaterial surfaces and presents a promising proof of concept for the use of ambient MS in the rapid and automated analysis of surface proteins.

  5. Adsorbed Water Illustration

    NASA Technical Reports Server (NTRS)

    2008-01-01

    The Thermal and Electrical Conductivity Probe on NASA's Phoenix Mars Lander detected small and variable amounts of water in the Martian soil.

    In this schematic illustration, water molecules are represented in red and white; soil minerals are represented in green and blue. The water, neither liquid, vapor, nor solid, adheres in very thin films of molecules to the surfaces of soil minerals. The left half illustrates an interpretation of less water being adsorbed onto the soil-particle surface during a period when the tilt, or obliquity, of Mars' rotation axis is small, as it is in the present. The right half illustrates a thicker film of water during a time when the obliquity is greater, as it is during cycles on time scales of hundreds of thousands of years. As the humidity of the atmosphere increases, more water accumulates on mineral surfaces. Thicker films behave increasingly like liquid water.

    The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.

  6. Fabrication Of Nano-Silver Thin Films Using Self Assembly And Its Interaction With Proteins

    SciTech Connect

    Verma, Gunjan; Choudhury, Sipra; Hassan, P. A.

    2010-12-01

    The silver nanoparticle thin films were prepared with an aim to use them for sensing of biomolecules. The monolayers of arachidic acid were deposited on glass plates by Langmuir Blodgett (LB) technique and silver nanoparticles thin films were deposited within the arachidic acid films. Small angle XRD studies confirm the formation of ordered array of nanoparticles. These thin films were treated with a model protein, bovine serum albumin (BSA a natural protein). From the optical absorption spectra a shift in the intensity as well as lambda max ({lambda}max) could be observed when silver thin films were treated with BSA.

  7. Effect of the incorporation of antimicrobial/antioxidant proteins on the properties of potato starch films.

    PubMed

    Moreno, Olga; Atarés, Lorena; Chiralt, Amparo

    2015-11-20

    Glycerol plasticized potato starch films containing bioactive proteins (lactoferrin (LF) and/or lysozyme (LZ), at 0.1 and 0.2 ratio with respect to starch) were obtained by casting method and characterized as to their microstructural, thermal and physical (water content, mechanical, water and oxygen barrier, optical) properties. The bioactive properties, named antioxidant and antimicrobial, of the proteins and the films were also characterized. The incorporation of proteins affected the structural and physical properties of potato starch films, while modifying their thermal behavior and increasing the glass transition temperature. Both proteins showed a certain degree of compatibility with starch chains through the bond formations (increase in Tg), while a part is separated and migrates to the film surface. Their incorporation, especially that of lactoferrin, greatly increased the film's brittleness, regardless of the films water content, although they enhance the water vapor and oxygen barrier properties, whatever the age of the film. Protein also reduced the film's transparency and gloss, while lactoferrin induced color changes. The thermal degradation of blend films and isolated proteins occurred at temperatures of over 250°C, which means that blend starch films can be thermoprocessed, according to their thermoplastic properties and following the usual practices of the plastics industries. A synergistic antimicrobial action against Escherichia coli and coliforms was observed when both LZ and LF were simultaneously applied. Both of these exhibited antioxidant capacity.

  8. Properties of cast films from hemp (Cannabis sativa L.) and soy protein isolates. A comparative study.

    PubMed

    Yin, Shou-Wei; Tang, Chuan-He; Wen, Qi-Biao; Yang, Xiao-Quan

    2007-09-05

    The properties of cast films from hemp protein isolate (HPI) including moisture content (MC) and total soluble mass (TSM), tensile strength (TS) and elongation at the break (EAB), and surface hydrophobicity were investigated and compared to those from soy protein isolate (SPI). The plasticizer (glycerol) level effect on these properties and the interactive force pattern for the film network formation were also evaluated. At some specific glycerol levels, HPI films had similar MC, much less TSM and EAB, and higher TS and surface hydrophobicity (support matrix side), as compared to SPI films. The TS of HPI and SPI films as a function of plasticizer level (in the range of 0.3-0.6 g/g of protein) were well fitted with the exponential equation with coefficient factors of 0.991 and 0.969, respectively. Unexpectedly, the surface hydrophobicity of HPI films (including air and support matrix sides) increased with increasing the glycerol level (from 0.3 to 0.6 g/g of protein). The analyses of protein solubility of film in various solvents and free sulfydryl group content showed that the disulfide bonds are the prominent interactive force in the HPI film network formation, while in the SPI case, besides the disulfide bonds, hydrogen bonds and hydrophobic interactions are also to a similar extent involved. The results suggest that hemp protein isolates have good potential to be applied to prepare protein film with some superior characteristics, e.g., low solubility and high surface hydrophobicity.

  9. Processing of mussel adhesive protein analog thin films by matrix assisted pulsed laser evaporation

    NASA Astrophysics Data System (ADS)

    Cristescu, R.; Patz, T.; Narayan, R. J.; Menegazzo, N.; Mizaikoff, B.; Mihaiescu, D. E.; Messersmith, P. B.; Stamatin, I.; Mihailescu, I. N.; Chrisey, D. B.

    2005-07-01

    Mussel adhesive proteins are a new class of biologically-derived materials that possess unique biocompatibility, bioactivity, and adhesion properties. We have demonstrated successful thin film growth of 3,4-dihydroxyphenyl- L-alanine modified poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (DOPA modified- PEO-PPO-PEO) block copolymer, a mussel adhesive protein analog, using matrix assisted pulsed laser evaporation. We have demonstrated that the main functional groups of the mussel adhesive protein analog are present in the transferred film. The effect of increasing of chain length of the mussel adhesive protein analog on film structure was also examined. These novel polymer thin films could have numerous medical and technological applications if their thin film properties are similar to what is found in bulk. This is the first report of successful MAPLE deposition of this material as thin films.

  10. Degradation behavior of soy protein-wheat gluten films in simulated soil conditions.

    PubMed

    Park, S K; Hettiarachchy, N S; Were, L

    2000-07-01

    Films containing soy protein and wheat gluten were exposed to simulated farmland soil mix over a period of 30 days and monitored for degradation. The simulated farmland soil mix (topsoil/sand/Sunshine compost/vermiculite, 59:6:25:10, wt %) was mixed and stored at ambient humidity (48-55%) and temperature (20-24 degrees C); the soil mix was constantly maintained at 15% moisture by weight. Research focused on evaluating the effectiveness of gluten and cysteine additions on biodegradable behavior in the simulated farmland soil conditions. The four types of films, soy protein (S:G 1:0); soy protein with cysteine addition (S:G 1:0 + CYS); soy protein-wheat gluten (S:G 4:1); and soy protein-wheat gluten with cysteine addition (S:G 4:1 + CYS), were prepared at pH 7. 0 for degradation studies. Soy protein-gluten film rapidly degraded with 50% weight loss in about 10 days and with up to 95% weight loss in 30 days. Tensile strength and elongation of all soy protein-gluten films significantly decreased in 3 days. However, cysteine addition delayed the degradation rate of soy protein-gluten films. Soy protein-wheat gluten film disintegrated after 20 days in the simulated farmland soil environment. These results suggest that wheat gluten and cysteine addition to soy protein-based films could delay degradation rates due to their high disulfide contents.

  11. Protein-resistant polymer coatings based on surface-adsorbed poly(aminoethyl methacrylate)/poly(ethylene glycol) copolymers.

    PubMed

    Ionov, Leonid; Synytska, Alla; Kaul, Elisabeth; Diez, Stefan

    2010-01-11

    We report on the protein-resistant properties of glass substrates coated with novel copolymers of 2-aminoethyl methacrylate hydrochloride and poly(ethylene glycol) methyl ether methacrylate (AEM-PEG). In comparison to currently available protein-blocking polymer systems, such as poly-l-lysine-poly(ethylene glycol), silane-based poly(ethylene glycol), and poly(ethylene glycol) brushes prepared by surface-initiated polymerization, the proposed AEM-PEG offers the combined advantages of low cost, simplicity of use, and applicability in aqueous solutions. We demonstrate the capability of AEM-PEG to block the surface binding of globular proteins (tubulin), their assemblies (microtubules), and functional motor proteins (kinesin-1). Moreover, we demonstrate the applicability of AEM-PEG for surface patterning of proteins in microfluidic devices.

  12. Effects of human hair and nail proteins and their films on rat mast cells.

    PubMed

    Fujii, Toshihiro; Murai, Shinya; Ohkawa, Kousaku; Hirai, Toshihiro

    2008-06-01

    Human hair and nail are valuable materials for producing individual corresponding biocompatible materials. A rapid and convenient protein extraction method (Shindai method) and novel procedures for preparing their protein films from their extracts have been developed using human hair and nail. The effects of the human hair and nail proteins and their films on histamine release from rat peritoneal mast cells were investigated. Both protein solutions and their films, mainly consisting of keratins and matrix proteins, did not induce histamine release from the mast cells. Scanning electron microscopy (SEM) also showed that the mast cells were only slightly affected by adding the human hair and nail proteins or by incubating on their protein films. The IgE-dependent histamine release was inhibited by the hair and nail proteins and their films. Incubation of the mast cells with the hair and nail proteins prior to the addition of the IgE serum resulted in a high inhibition (50%) of the histamine release, while the inhibition was approximately 10% when the protein solutions were mixed with the mast cells after incubation with the IgE serum. These results suggest that the human hair and nail proteins and their films will be useful materials for antiallergic actions.

  13. Specific Interactions of Neutral Side Chains of an Adsorbed Protein with the Surface of α-Quartz and Silica Gel.

    PubMed

    Odinokov, Alexey V; Bagaturyants, Alexander A

    2015-07-16

    Many key features of the protein adsorption on the silica surfaces still remain unraveled. One of the open questions is the interaction of nonpolar side chains with siloxane cavities. Here, we use nonequilibrium molecular dynamics simulations for the detailed investigation of the binding of several hydrophobic and amphiphilic protein side chains with silica surface. These interactions were found to be a possible driving force for protein adsorption. The free energy gain was larger for the disordered surface of amorphous silica gel as compared to α-quartz, but the impact depended on the type of amino acid. The dependence was analyzed from the structural point of view. For every amino acid an enthalpy-entropy compensation behavior was observed. These results confirm a hypothesis of an essential role of hydrophobic interactions in protein unfolding and irreversible adsorption on the silica surface.

  14. Protein immobilization on epoxy-activated thin polymer films: effect of surface wettability and enzyme loading.

    PubMed

    Chen, Bo; Pernodet, Nadine; Rafailovich, Miriam H; Bakhtina, Asya; Gross, Richard A

    2008-12-02

    A series of epoxy-activated polymer films composed of poly(glycidyl methacrylate/butyl methacrylate/hydroxyethyl methacrylate) were prepared. Variation in comonomer composition allowed exploration of relationships between surface wettability and Candida antartica lipase B (CALB) binding to surfaces. By changing solvents and polymer concentrations, suitable conditions were developed for preparation by spin-coating of uniform thin films. Film roughness determined by AFM after incubation in PBS buffer for 2 days was less than 1 nm. The occurrence of single CALB molecules and CALB aggregates at surfaces was determined by AFM imaging and measurements of volume. Absolute numbers of protein monomers and multimers at surfaces were used to determine values of CALB specific activity. Increased film wettability, as the water contact angle of films increased from 420 to 550, resulted in a decreased total number of immobilized CALB molecules. With further increases in the water contact angle of films from 55 degrees to 63 degrees, there was an increased tendency of CALB molecules to form aggregates on surfaces. On all flat surfaces, two height populations, differing by more than 30%, were observed from height distribution curves. They are attributed to changes in protein conformation and/or orientation caused by protein-surface and protein-protein interactions. The fraction of molecules in these populations changed as a function of film water contact angle. The enzyme activity of immobilized films was determined by measuring CALB-catalyzed hydrolysis of p-nitrophenyl butyrate. Total enzyme specific activity decreased by decreasing film hydrophobicity.

  15. Wettability Effect of PECVD-SiOx Films on Poly(lactic acid) Induced by Oxygen Plasma on Protein Adsorption and Cell Attachment

    NASA Astrophysics Data System (ADS)

    Sarapirom, S.; Lee, J. S.; Jin, S. B.; Song, D. H.; Yu, L. D.; Han, J. G.; Chaiwong, C.

    2013-04-01

    Surface wettability is an important property of biomaterials. Silicon oxide films have a wide range of applications due to a range of the properties such as the mechanical strength and surface wettability. This paper reports effect of the surface wettability of silicon oxide (SiOx) films on protein adsorption and cell attachment and proliferation. SiOx films were deposited onto poly(lactic acid) (PLA) substrate using plasma enhanced chemical vapor deposition (PECVD). Octamethylcyclotetrasiloxane (OMCTS:Si4O4C8H24) was used as a precursor with O2 as a carrier gas. After deposition, the films were treated with O2-plasma to adapt wettability. It was found that O2-plasma enhanced the wettability of the films without changing the film thickness, while made the surface morphology slightly smoother. The polar component increased after O2-plasma treatment as observed in the contact angle measurements. The surface energy of the films was calculated by means of the Owens-Wendt method to resolve the contributions of polar and dispersive components. The chemical structure was characterized using attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy. The films were dense with a high Si-network structure. The reduced carbon content (-CHn, Si-CH3) and increased hydrogen content (-OH) of the O2-plasma treated SiOx films led to the polar components enhancing the SiOx wettability. Adsorption of bovine serum albumin (BSA) on the films was investigated by using x-ray photoelectron spectroscopy (XPS). More BSA was adsorbed onto the O2-plasma treated SiOx films. Attachment and proliferation of MC3T3-E1 mouse pre-osteoblasts and L929 mouse fibroblasts cells on the SiOx films were evaluated via MTT assay. The cells were attached more to the untreated SiOx films but proliferated more on the surface of the O2-plasma treated SiOx films depending on the cell types.

  16. Mechanical and thermal properties of irradiated films based on Tilapia ( Oreochromis niloticus) proteins

    NASA Astrophysics Data System (ADS)

    Sabato, S. F.; Nakamurakare, N.; Sobral, P. J. A.

    2007-11-01

    Proteins are considered potential material in natural films as alternative to traditional packaging. When gamma radiation is applied to protein film forming solution it resulted in an improvement in mechanical properties of whey protein films. The objective of this work was the characterization of mechanical and thermal properties of irradiated films based on muscle proteins from Nile Tilapia ( Oreochromis niloticus). The films were prepared according to a casting technique with two levels of plasticizer: 25% and 45% glycerol and irradiated in electron accelerator type Radiation Dynamics, 0.550 MeV at dose range from 0 to 200 kGy. Thermal properties and mechanical properties were determined using a differential scanning calorimeter and a texture analyzer, respectively. Radiation from electron beam caused a slightly increase on its tensile strength characteristic at 100 kGy, while elongation value at this dose had no reduction.

  17. Fungal Hydrophobin Proteins Produce Self-Assembling Protein Films with Diverse Structure and Chemical Stability

    PubMed Central

    Lo, Victor C.; Ren, Qin; Pham, Chi L. L.; Morris, Vanessa K.; Kwan, Ann H.; Sunde, Margaret

    2014-01-01

    Hydrophobins are small proteins secreted by fungi and which spontaneously assemble into amphipathic layers at hydrophilic-hydrophobic interfaces. We have examined the self-assembly of the Class I hydrophobins EAS∆15 and DewA, the Class II hydrophobin NC2 and an engineered chimeric hydrophobin. These Class I hydrophobins form layers composed of laterally associated fibrils with an underlying amyloid structure. These two Class I hydrophobins, despite showing significant conformational differences in solution, self-assemble to form fibrillar layers with very similar structures and require a hydrophilic-hydrophobic interface to trigger self-assembly. Addition of additives that influence surface tension can be used to manipulate the fine structure of the protein films. The Class II hydrophobin NC2 forms a mesh-like protein network and the engineered chimeric hydrophobin displays two multimeric forms, depending on assembly conditions. When formed on a graphite surface, the fibrillar EAS∆15 layers are resistant to alcohol, acid and basic washes. In contrast, the NC2 Class II monolayers are dissociated by alcohol treatment but are relatively stable towards acid and base washes. The engineered chimeric Class I/II hydrophobin shows increased stability towards alcohol and acid and base washes. Self-assembled hydrophobin films may have extensive applications in biotechnology where biocompatible; amphipathic coatings facilitate the functionalization of nanomaterials.

  18. Uremic toxins and oral adsorbents.

    PubMed

    Goto, Shunsuke; Yoshiya, Kunihiko; Kita, Tomoyuki; Fujii, Hideki; Fukagawa, Masafumi

    2011-04-01

    Uremic toxins are associated with various disorders in patients with end-stage renal disease and it is difficult to remove some of these toxins by dialysis. Since some uremic toxins are generated by bacterial metabolites in the colon, oral adsorbents that interfere with the absorption of uremic toxins or their precursors are believed to prevent their accumulation in the body. AST-120 adsorbs various uremic retention solutes in the gastrointestinal system and has potential for providing clinical benefit. Sevelamer hydrochloride binds some harmful compounds in addition to phosphate and seems to have pleiotropic effects that include lowering serum LDL cholesterol levels and reduction of inflammation. The effect of sevelamer hydrochloride on indoxyl sulfate and p-cresol has been shown in an in vitro study; however, in vivo studies in mice or humans did not demonstrate this effect on protein-binding uremic toxins. Oral adsorbents are thus one of the important modalities in the treatment of uremic syndrome.

  19. Physical properties of whey protein coating solutions and films containing antioxidants.

    PubMed

    Han, J H; Krochta, J M

    2007-06-01

    Antioxidants (ascorbyl palmitate and alpha-tocopherol) were incorporated into 10% (w/w) whey protein isolate (WPI) coating solution containing 6.67% (w/w) glycerol (WPI:glycerol = 6:4). Before incorporation, the antioxidants were mixed using either powder blending (Process 1) or ethanol solvent-mixing (Process 2). After the antioxidant mixtures were incorporated into heat-denatured WPI solution, viscosity and turbidity of the WPI solutions were determined. The WPI solutions were dried on a flat surface to produce WPI films. The WPI films were examined to determine transparency and oxygen-barrier properties (permeability, diffusivity, and solubility). WPI solution containing antioxidants produced by Process 1 and Process 2 did not show any difference in viscosity and turbidity, but viscosity was greater for the WPI solution with rather than without antioxidants. WPI films produced by Process 2 were more transparent than the films produced by Process 1. Oxygen permeability of Process 1 film was lower than Process 2 film. However, both the diffusivity and solubility of oxygen were statistically the same in Process 1 and Process 2 films. Both control WPI films and antioxidant-containing WPI films had very low oxygen solubility, comparable to polyethylene terephthalate films. Permeability of antioxidant-incorporated films was not enhanced compared to control WPI films.

  20. Hydrophobicity, thermal and micro-structural properties of whey protein concentrate-pullulan-beeswax films.

    PubMed

    Jafari, Seid Mahdi; Khanzadi, Mehrdad; Mirzaei, Habibollah; Dehnad, Danial; Chegini, Faramarz Khodaian; Maghsoudlou, Yayha

    2015-09-01

    In this research, effects of beeswax (BW) on functional properties of whey protein concentrates (WPC):pullulan (PUL) films were investigated. For this purpose, 0, 10, 20 and 30w/w(glycerol)% BW rates and 30:70, 50:50 and 70:30w/w% WPC:PUL ratios were applied. Films containing 70% WPC:30% PUL (WPC70) and 30% BW (BW30) justified the highest contact angle (92.4°) among all films; SEM micrographs indicated that BW could come toward the surface of films during drying stage and resulted in a higher hydrophobic behavior of bilayer films compared with blend films. WPC70 supplied the lowest T(g) values (36-48 °C) among different proportions of WPC-PUL; the highest melting points were just assured in the absence of BW regardless of combination ratio for WPI:PUL. BW30 films deserved lower roughness rates than BW20 (and even BW10) films, indicating more advantageous microstructure and higher hydrogen connections in BW30 films and justifying similar melting points attained for BW30 films to BW20 or 10 ones. Overall, application of WPC70 and BW30 was recommended to obtain optimum combination of final properties for WPC-PUL-BW bilayer films as SEM exhibited flexible and elastic structures of such films.

  1. From art to science in protein crystallization by means of thin-film nanotechnology

    NASA Astrophysics Data System (ADS)

    Pechkova, Eugenia; Nicolini, Claudio

    2002-08-01

    A new method of protein crystallization based on a homologous Langmuir-Schaefer (LS) protein thin-film template proves to successfully stimulate crystal nucleation and growth of the four different proteins being studied, under different crystallization conditions including those failing to lead to crystal formation in solution. Protein microcrystals were obtained by a modified vapour diffusion hanging drop method. This report is focused on recombinant bovine cytochrome P-450scc crystallization and subsequent crystal characterization by atomic force microscopy in an appropriate chamber. The results are discussed in terms of a possible transition in protein crystallization from art to science by means of LS thin-film nanotechnology.

  2. Partial high-resolution structure of phosphorylated and non-phosphorylated leucine-rich amelogenin protein adsorbed to hydroxyapatite

    SciTech Connect

    Masica, David L.; Gray, Jeffrey J.; Shaw, Wendy J.

    2011-07-21

    The formation of biogenic materials requires the interaction of organic molecules with the mineral phase. In forming enamel, the amelogenin proteins contribute to the mineralization of hydroxyapatite (HAp). Leucine-rich amelogenin protein (LRAP) is a naturally occurring splice variant of amelogenin that comprises amelogenin’s predicted HAp binding domains. We determined the partial structure of phosphorylated and non-phosphorylated LRAP variants bound to HAp using combined solid-state NMR (ssNMR) and ssNMR-biased computational structure prediction. The ssNMR measurements indicate a largely extended structure for both variants, though some measurements are consistent with a partially helical N-terminal segment. Structure prediction was biased using 21 ssNMR measurements at five HAp crystal faces. The predicted fold of LRAP is similar at all HAp faces studied, regardless of phosphorylation. LRAP’s predicted structure is relatively extended with a helix-turn-helix motif in the N-terminal domain and some helix in the C-terminal domain. The N-terminal domain of the phosphorylated variant binds HAp more tightly than the N-terminal domain of the non-phosphorylated variant. Both variants are predicted to preferentially bind the {010} HAp crystal face providing further evidence that amelogenins block crystal growth on the a and b faces to allow elongated crystals in the c-axis. Pacific Northwest National Laboratory is operated by Battelle for the US Department of Energy.

  3. Force measurements on myelin basic protein adsorbed to mica and lipid bilayer surfaces done with the atomic force microscope.

    PubMed Central

    Mueller, H; Butt, H J; Bamberg, E

    1999-01-01

    The mechanical and adhesion properties of myelin basic protein (MBP) are important for its function, namely the compaction of the myelin sheath. To get more information about these properties we used atomic force microscopy to study tip-sample interaction of mica and mixed dioleoylphosphatidylserine (DOPS) (20%)/egg phosphatidylcholine (EPC) (80%) lipid bilayer surfaces in the absence and presence of bovine MBP. On mica or DOPS/EPC bilayers a short-range repulsive force (decay length 1.0-1.3 nm) was observed during the approach. The presence of MBP always led to an attractive force between tip and sample. When retracting the tip again, force curves on mica and on lipid layers were different. While attached to the mica surface, the MBP molecules exhibited elastic stretching behavior that agreed with the worm-like chain model, yielding a persistence length of 0.5 +/- 0.25 nm and an average contour length of 53 +/- 19 nm. MBP attached to a lipid bilayer did not show elastic stretching behavior. This shows that the protein adopts a different conformation when in contact with lipids. The lipid bilayer is strongly modified by MBP attachment, indicating formation of MBP-lipid complexes and possibly disruption of the original bilayer structure. PMID:9916039

  4. Denaturation resistance of beta-lactoglobulin in monomolecular films at the air-water interface.

    PubMed

    Lin, Jhih-Min; White, John W

    2009-10-29

    Using X-ray reflectometry we report strong differences in the denaturation response of beta-lactoglobulin adsorbed as a monomolecular film at the air-water interface from that observed in mixed denaturant/beta-lactoglobulin bulk solutions. Using the "flow trough" technique an isolated monomolecular film of the protein showed little change in structure when subjected to a 4.0 M guanidinium hydrochloride substrate. Unlike the bulk solution where a new protein layer structure appears, small changes in the protein packing and the roughness of the film are the only evidence of change. These parameters have been studied as a function of denaturant concentration and film quality. The strength of the response depends on the degree of perfection of the originally formed film; quickly formed films are more easily denatured. As the response is so subtle, possible interfering effects such as denaturant release of protein adsorbed on the trough have been quantified.

  5. Physical, Chemical and Biochemical Modifications of Protein-Based Films and Coatings: An Extensive Review

    PubMed Central

    Zink, Joël; Wyrobnik, Tom; Prinz, Tobias; Schmid, Markus

    2016-01-01

    Protein-based films and coatings are an interesting alternative to traditional petroleum-based materials. However, their mechanical and barrier properties need to be enhanced in order to match those of the latter. Physical, chemical, and biochemical methods can be used for this purpose. The aim of this article is to provide an overview of the effects of various treatments on whey, soy, and wheat gluten protein-based films and coatings. These three protein sources have been chosen since they are among the most abundantly used and are well described in the literature. Similar behavior might be expected for other protein sources. Most of the modifications are still not fully understood at a fundamental level, but all the methods discussed change the properties of the proteins and resulting products. Mastering these modifications is an important step towards the industrial implementation of protein-based films. PMID:27563881

  6. Properties of blend film based on cuttlefish (Sepia pharaonis) skin gelatin and mungbean protein isolate.

    PubMed

    Hoque, Md Sazedul; Benjakul, Soottawat; Prodpran, Thummanoon; Songtipya, Ponusa

    2011-11-01

    Blend films based on cuttlefish (Sepia pharaonis) ventral skin gelatin (CG) and mungbean protein isolate (MPI) at different blend ratios (CG/MPI=10:0, 8:2, 6:4, 4:6, 2:8 and 0:10, w/w) prepared at pH 11 using 50% glycerol (based on total protein) as plasticizer were characterized. CG films incorporated with MPI at increasing amounts had the decreases in tensile strength (TS) (p<0.05). The increases in elongation at break (EAB) were observed when CG/MPI ratios of 6:4 or 4:6 were used (p<0.05). Decreased water vapor permeability (WVP) was obtained for films having the increasing proportion of MPI (p<0.05). CG/MPI blend films with higher MPI proportion had lower film solubility and L*-values (lightness) but higher b*-values (yellowness) and ΔE*-values (total color difference) (p<0.05). Electrophoretic study revealed that disulfide bond was present in MPI and CG/MPI blend films. However, hydrogen bonds between CG and MPI in the film matrix were dominant, as elucidated from FTIR spectroscopic analysis. Moreover, thermal stability of CG/MPI blend film was improved as compared to that of films from respective single proteins. Differential scanning calorimetry result suggested solid-state morphology of CG/MPI (6:4) blend film that consisted of amorphous phase of partially miscible CG/MPI mixture and the coexisting two different order phases of individual CG and MPI domains. Thus, the incorporation of MPI into gelatin film could improve the properties of resulting blend film, which were governed by CG/MPI ratio.

  7. Incorporation and thermal evolution of rhodamine 6G dye molecules adsorbed in porous columnar optical SiO2 thin films.

    PubMed

    Sánchez-Valencia, Juan R; Blaszczyk-Lezak, Iwona; Espinós, Juan P; Hamad, Said; González-Elipe, Agustín R; Barranco, Angel

    2009-08-18

    Rhodamine 6G (Rh6G) dye molecules have been incorporated into transparent and porous SiO2 thin films prepared by evaporation at glancing angles. The porosity of these films has been assessed by analyzing their water adsorption isotherms measured for the films deposited on a quartz crystal monitor. Composite Rh6G/SiO2 thin films were prepared by immersion of a SiO2 thin film into a solution of the dye at a given pH. It is found that the amount of Rh6G molecules incorporated into the film is directly dependent on the pH of the solution and can be accounted for by a model based on the point of zero charge (PZC) concepts originally developed for colloidal oxides. At low pHs, the dye molecules incorporate in the form of monomers, while dimers or higher aggregates are formed if the pH increases. Depending on the actual preparation and treatment conditions, they also exhibit high relative fluorescence efficiency. The thermal stability of the composite films has been also investigated by characterizing their optical behavior after heating in an Ar atmosphere at increasing temperatures up to 275 degrees C. Heating induces a progressive loss of active dye molecules, a change in their agglomeration state, and an increment in their relative fluorescence efficiency. The obtained Rh6G/SiO2 composite thin films did not disperse the light and therefore can be used for integration into optical and photonic devices.

  8. Identification of polymer surface adsorbed proteins implicated in pluripotent human embryonic stem cell expansion† †Electronic supplementary information (ESI) available. See DOI: 10.1039/c6bm00214e Click here for additional data file.

    PubMed Central

    Hammad, Moamen; Rao, Wei; Smith, James G. W.; Anderson, Daniel G.; Langer, Robert; Young, Lorraine E.; Barrett, David A.; Davies, Martyn C.; Denning, Chris

    2016-01-01

    Improved biomaterials are required for application in regenerative medicine, biosensing, and as medical devices. The response of cells to the chemistry of polymers cultured in media is generally regarded as being dominated by proteins adsorbed to the surface. Here we use mass spectrometry to identify proteins adsorbed from a complex mouse embryonic fibroblast (MEF) conditioned medium found to support pluripotent human embryonic stem cell (hESC) expansion on a plasma etched tissue culture polystyrene surface. A total of 71 proteins were identified, of which 14 uniquely correlated with the surface on which pluripotent stem cell expansion was achieved. We have developed a microarray combinatorial protein spotting approach to test the potential of these 14 proteins to support expansion of a hESC cell line (HUES-7) and a human induced pluripotent stem cell line (ReBl-PAT) on a novel polymer (N-(4-Hydroxyphenyl) methacrylamide). These proteins were spotted to form a primary array yielding several protein mixture ‘hits’ that enhanced cell attachment to the polymer. A second array was generated to test the function of a refined set of protein mixtures. We found that a combination of heat shock protein 90 and heat shock protein-1 encourage elevated adherence of pluripotent stem cells at a level comparable to fibronectin pre-treatment. PMID:27466628

  9. Dynamic Contact Angle Analysis of Protein Adsorption on Polysaccharide Multilayer's Films for Biomaterial Reendothelialization

    PubMed Central

    Benni, Safiya; Mora, Laurence

    2014-01-01

    Atherosclerosis is a major cardiovascular disease. One of the side effects is restenosis. The aim of this work was to study the coating of stents by dextran derivates based polyelectrolyte's multilayer (PEM) films in order to increase endothelialization of injured arterial wall after stent implantation. Films were composed with diethylaminoethyl dextran (DEAE) as polycation and dextran sulphate (DS) as polyanion. One film was composed with 4 bilayers of (DEAE-DS)4 and was labeled D−. The other film was the same as D− but with an added terminal layer of DEAE polycation: (DEAE-DS)4-DEAE (labeled D+). The dynamic adsorption/desorption of proteins on the films were characterized by dynamic contact angle (DCA) and atomic force microscopy (AFM). Human endothelial cell (HUVEC) adhesion and proliferation were quantified and correlated to protein adsorption analyzed by DCA for fibronectin, vitronectin, and bovine serum albumin (BSA). Our results showed that the endothelial cell response was optimal for films composed of DS as external layer. Fibronectin was found to be the only protein to exhibit a reversible change in conformation after desorption test. This behavior was only observed for (DEAE-DS)4 films. (DEAE-DS)4 films could enhance HUVEC proliferation in agreement with fibronectin ability to easily change from conformation. PMID:25276808

  10. Dynamic contact angle analysis of protein adsorption on polysaccharide multilayer's films for biomaterial reendothelialization.

    PubMed

    Benni, Safiya; Avramoglou, Thierry; Hlawaty, Hanna; Mora, Laurence

    2014-01-01

    Atherosclerosis is a major cardiovascular disease. One of the side effects is restenosis. The aim of this work was to study the coating of stents by dextran derivates based polyelectrolyte's multilayer (PEM) films in order to increase endothelialization of injured arterial wall after stent implantation. Films were composed with diethylaminoethyl dextran (DEAE) as polycation and dextran sulphate (DS) as polyanion. One film was composed with 4 bilayers of (DEAE-DS)4 and was labeled D-. The other film was the same as D- but with an added terminal layer of DEAE polycation: (DEAE-DS)4-DEAE (labeled D+). The dynamic adsorption/desorption of proteins on the films were characterized by dynamic contact angle (DCA) and atomic force microscopy (AFM). Human endothelial cell (HUVEC) adhesion and proliferation were quantified and correlated to protein adsorption analyzed by DCA for fibronectin, vitronectin, and bovine serum albumin (BSA). Our results showed that the endothelial cell response was optimal for films composed of DS as external layer. Fibronectin was found to be the only protein to exhibit a reversible change in conformation after desorption test. This behavior was only observed for (DEAE-DS)4 films. (DEAE-DS)4 films could enhance HUVEC proliferation in agreement with fibronectin ability to easily change from conformation.

  11. Characterization of whey protein-carboxymethylated chitosan composite films with and without transglutaminase treatment.

    PubMed

    Jiang, Shu-Juan; Zhang, Xuan; Ma, Ying; Tuo, Yanfeng; Qian, Fang; Fu, Wenjia; Mu, Guangqing

    2016-11-20

    Edible composite packaging has the advantage of complementary functional properties over its each bio-components. However, reports on whey protein concentrates (WPC)-carboxymethylated chitosan (CMC) composite films have not yet been released. To investigate the preparation of WPC-CMC composite films and its functional properties, four types of WPC-CMC composite films were prepared with and without Transglutaminase (TGase) treatment by mixing WPC aqueous solutions (10%, w/v) with CMC aqueous solutions (3%, w/v) at WPC to CMC volume ratios of (100:0), (75:25), (50:50), and (25:75). SDS-PAGE confirmed that TGase catalyzed crosslinking of whey protein. Results revealed that CMC incorporation conferred a smooth and even surface microstructure on the films and markedly improved the transparency, water barrier properties, mechanical properties and solubility of the composite film. Furthermore, TGase resulted in an improvement in the water vapor barrier properties and mechanical properties of WPC-CMC (75:25 and 50:50, v/v) composite films, and there was no impairment of thermal stability of composite films. Therefore, TGase successfully facilitated the formation of WPC-CMC composite films with some improved functional properties. This offers potential applications as an alternative approach to the preparation of edible packaging films.

  12. Modeling adsorption: Investigating adsorbate and adsorbent properties

    NASA Astrophysics Data System (ADS)

    Webster, Charles Edwin

    1999-12-01

    Surface catalyzed reactions play a major role in current chemical production technology. Currently, 90% of all chemicals are produced by heterogeneously catalyzed reactions. Most of these catalyzed reactions involve adsorption, concentrating the substrate(s) (the adsorbate) on the surface of the solid (the adsorbent). Pore volumes, accessible surface areas, and the thermodynamics of adsorption are essential in the understanding of solid surface characteristics fundamental to catalyst and adsorbent screening and selection. Molecular properties such as molecular volumes and projected molecular areas are needed in order to convert moles adsorbed to surface volumes and areas. Generally, these molecular properties have been estimated from bulk properties, but many assumptions are required. As a result, different literature values are employed for these essential molecular properties. Calculated molar volumes and excluded molecular areas are determined and tabulated for a variety of molecules. Molecular dimensions of molecules are important in the understanding of molecular exclusion as well as size and shape selectivity, diffusion, and adsorbent selection. Molecular dimensions can also be used in the determination of the effective catalytic pore size of a catalyst. Adsorption isotherms, on zeolites, (crystalline mineral oxides) and amorphous solids, can be analyzed with the Multiple Equilibrium Analysis (MEA) description of adsorption. The MEA produces equilibrium constants (Ki), capacities (ni), and thermodynamic parameters (enthalpies, ΔHi, and entropies, ΔSi) of adsorption for each process. Pore volumes and accessible surface areas are calculated from the process capacities. Adsorption isotherms can also be predicted for existing and new adsorbate-adsorbent systems with the MEA. The results show that MEA has the potential of becoming a standard characterization method for microporous solids that will lead to an increased understanding of their behavior in gas

  13. Preparation of a porcine plasma protein composite film and its application.

    PubMed

    Lee, Ji-Hyun; Song, Kyung Bin

    2015-01-01

    To use blood released from slaughtering houses, a porcine plasma protein (PPP)/nanoclay composite film was prepared. The tensile strength and elongation at break values of the PPP composite film with 5% nanoclay were 10.01 MPa and 6.55%, respectively. The PPP composite film containing 1% grapefruit seed extract (GSE) was applied to pork meat, and the populations of inoculated Escherichia coli O157:H7 and Listeria monocytogenes in the pork meat packaged with the PPP composite film decreased by 0.8 and 1.0 log CFU/g, respectively, after 7 days of storage compared to the populations of the control. In addition, thiobarbituric acid values in the pork meat packaged with the PPP composite film were less than those of the control sample during storage. These results suggest that the PPP nanocomposite film containing 1% GSE can be used as a packaging material to maintain the quality of pork meat.

  14. Oral films as breakthrough tools for oral delivery of proteins/peptides.

    PubMed

    Castro, Pedro M; Fonte, Pedro; Sousa, Flávia; Madureira, Ana Raquel; Sarmento, Bruno; Pintado, Manuela E

    2015-08-10

    Therapeutic proteins and peptides demonstrate unique, peerless, pharmacological characteristics such as high specificity to receptors and superior biological mimicking of physiological mechanisms, resulting in a better therapeutic index compared to conventional chemical-derived drugs. However, proteins also present inherent bioavailability limitations. Thus, this paper proposes several effective tools to improve protein/peptide drugs stability, permeability and pharmacokinetics with special emphasis on oral polymeric films as oral delivery platforms. Indeed, oral films present inherent characteristics that can greatly enhance biological performance of proteins and peptides and patient compliance along with other advantages that are critically discussed in this review. A rational choice of excipients addressed in and manufacture processes are also focused. In addition, possible toxicity issues to be overtaken and critical analysis regarding current market tendencies respecting oral films and protein/peptides along with future prospects are disclosed.

  15. Infrared Reflection-Absorption Spectroscopy: Principles and Applications to Lipid-Protein Interaction in Langmuir Films

    PubMed Central

    Mendelsohn, Richard; Mao, Guangru; Flach, Carol R.

    2010-01-01

    Infrared reflection-absorption spectroscopy (IRRAS) of lipid/protein monolayer films in situ at the air/water interface provides unique molecular structure and orientation information from the film constituents. The technique is thus well suited for studies of lipid/protein interaction in a physiologically relevant environment. Initially, the nature of the IRRAS experiment is described and the molecular structure information that may be obtained is recapitulated. Subsequently, several types of applications, including the determination of lipid chain conformation and tilt as well as elucidation of protein secondary structure are reviewed. The current article attempts to provide the reader with an understanding of the current capabilities of IRRAS instrumentation and the type of results that have been achieved to date from IRRAS studies of lipids, proteins and lipid/protein films of progressively increasing complexity. Finally, possible extensions of the technology are briefly considered. PMID:20004639

  16. High-pressure homogenization lowers water vapor permeability of soybean protein isolate-beeswax films.

    PubMed

    Zhang, Chao; Ma, Yue; Guo, Kuan; Zhao, Xiaoyan

    2012-03-07

    Soybean-protein isolate (SPI) has excellent film-forming capacity. However, the water vapor permeability of SPI film is high, which will cause the moisture lose of packaged products. The effect of high-pressure homogenization (HPH) on the water vapor permeability of SPI-beeswax films was evaluated. The HPH was effective at lowering the water vapor permeability of SPI-beeswax films to about 50% of the control. The HPH reduced the particle size of films and made their matrix more compact. The HPH improved the hydrophobicity of SPI-beeswax films. For the first time, we proved that the HPH improved the bound-beeswax content in SPI-beeswax films. The bound beeswax was effective at lowering the water vapor permeability of films rather than the free beeswax in the film matrix. In summary, the HPH lowered water vapor permeability of SPI-beeswax films by reducing their particle size and raising their hydrophobicity and bound-beeswax content.

  17. Protein microarrays on hybrid polymeric thin films prepared by self-assembly of polyelectrolytes for multiple-protein immunoassays.

    PubMed

    Zhou, Xichun; Zhou, Jizhong

    2006-03-01

    We report here the development and characterization of protein microarrays fabricated on nanoengineered 3-D polyelectrolyte thin films (PET) deposited on glass slide by consecutive adsorption of polyelectrolytes via self-assembly technique. Antibodies or antigens were immobilized in the PET-coated glass slides by electrostatic adsorption and entrapment of porous structure of the 3-D polymer film and thus establishing a platform for parallel analysis. Both antigen and antibody microarrays were fabricated on the PET-coated slides, and direct and indirect immunoassays on protein microarrays for multiple-analyte detection were demonstrated. Microarrays produced on these PET-coated slides have consistent spot morphology and provide performance features needed for proteomic analysis. The protein microarrays on the PET films provide LOD as low as 6 pg/mL and dynamic ranges up to three orders of magnitude, which are wider than the protein microarrays fabricated on aldehyde and poly-L-lysine functionalized slides. The PET films constructed by self-assembly technique in aqueous solution is green chemistry based, cost-effective method to generate 3-D thin film coatings on glass surface, and the coated slide is well suited for immobilizing many types of biological molecules so that a wide variety of microarray formats can be developed on this type of slide.

  18. Effect of protein and glycerol concentration on the mechanical, optical, and water vapor barrier properties of canola protein isolate-based edible films.

    PubMed

    Chang, Chang; Nickerson, Michael T

    2015-01-01

    Biodegradable edible films prepared using proteins are both economically and environmentally important to the food packaging industry relative to traditional petroleum-derived synthetic materials. In the present study, the mechanical and water vapor barrier properties of casted canola protein isolate edible films were investigated as a function of protein (5.0% and 7.5%) and glycerol (30%, 35%, 40%, 45%, and 50%) content. Specifically, tensile strength and elongation, elastic modulus, puncture strength and deformation, opacity, and water vapor permeability were measured. Results indicated that tensile strength, puncture strength, and elastic modulus decreased, while tensile elongation and puncture deformation values increased as glycerol concentration increased for both 5.0% and 7.5% canola protein isolate films. Furthermore, tensile strength, puncture strength, and elastic modulus values were found to increase at higher protein concentrations within the canola protein isolate films, whereas puncture deformation values decreased. Tensile elongation was found to be similar for both canola protein isolate protein levels. Canola protein isolate films became more transparent with increasing of glycerol concentration and decreasing of canola protein isolate concentration. Water vapor permeability value was also found to increase with increasing glycerol and protein contents. Overall, results indicated that canola protein isolate films were less brittle, more malleable and transparent, and had greater water vapor permeability at higher glycerol levels. However, as protein level increased, canola protein isolate films were more brittle, less malleable and more opaque, and also had increased water vapor permeability.

  19. Effect of transglutaminase on properties of tilapia scale gelatin films incorporated with soy protein isolate.

    PubMed

    Weng, Wuyin; Zheng, Huibin

    2015-02-15

    The effect of transglutaminase (TGase) on the properties of tilapia scale gelatin films in the presence of soy protein isolate (SPI) was investigated. When 3% TGase was added into gelatin films, the total soluble matter and protein solubility of films were decreased from 89.36% and 92.78% to 35.83% and 40.05%, respectively, and the decline was promoted by adding 5% SPI. The strength of the films was increased by adding 1% TGase irrespective of SPI addition, but decreased when the TGase concentration was further raised. No obvious colour change was observed in the films with or without TGase and SPI. Based on the results of SDS-PAGE, DSC and SEM, it was revealed that the movement of low molecular weight hydrophilic protein was depressed by the cross-linking network structure induced by TGase and SPI during film drying, indicating that adding SPI is essential to improve the thermal stability and water resistance properties of TGase-induced gelatin films.

  20. Ultralow protein adsorbing coatings from clickable PEG nanogel solutions: Benefits of attachment under salt-induced phase separation conditions and comparison with PEG/albumin nanogel coatings

    PubMed Central

    Donahoe, Casey D.; Cohen, Thomas L.; Li, Wenlu; Nguyen, Peter K.; Fortner, John D.; Mitra, Robi D.; Elbert, Donald L.

    2013-01-01

    Clickable nanogel solutions were synthesized by using the copper catalyzed azide/alkyne cycloaddition (CuAAC) to partially polymerize solutions of azide and alkyne functionalized poly(ethylene glycol) (PEG) monomers. Coatings were fabricated using a second click reaction: a UV thiol-yne attachment of the nanogel solutions to mercaptosilanated glass. Because the CuAAC reaction was effectively halted by the addition of a copper-chelator, we were able to prevent bulk gelation and limit the coating thickness to a single monolayer of nanogels in the absence of the solution reaction. This enabled the inclusion of kosmotropic salts, which caused the PEG to phase-separate and nearly double the nanogel packing density, as confirmed by Quartz Crystal Microbalance with Dissipation (QCM-D). Protein adsorption was analyzed by single molecule counting with total internal reflection fluorescence (TIRF) microscopy and cell adhesion assays. Coatings formed from the phase-separated clickable nanogel solutions attached with salt adsorbed significantly less fibrinogen than other 100% PEG coatings tested, as well as poly-L-lysine-g-PEG (PLL-g-PEG) coatings. However, PEG/albumin nanogel coatings still outperformed the best 100% PEG clickable nanogel coatings. Additional surface crosslinking of the clickable nanogel coating in the presence of copper further reduced levels of fibrinogen adsorption closer to those of PEG/albumin nanogel coatings. However, this step negatively impacted long-term resistance to cell adhesion and dramatically altered the morphology of the coating by atomic force microscopy (AFM). The main benefit of the click strategy is that the partially polymerized solutions are stable almost indefinitely, allowing attachment in the phase-separated state without danger of bulk gelation, and thus, producing the best performing 100% PEG coating that we have studied to date. PMID:23441808

  1. Anion exchange membrane adsorbers for flow-through polishing steps: Part II. Virus, host cell protein, DNA clearance, and antibody recovery.

    PubMed

    Weaver, Justin; Husson, Scott M; Murphy, Louise; Wickramasinghe, S Ranil

    2013-02-01

    Anion exchange membrane adsorbers are used for contaminant removal in flow-through polishing steps in the manufacture of biopharmaceuticals. This contribution describes the clearance of minute virus of mice, DNA, and host cell proteins by three commercially available anion-exchange membranes: Sartobind Q, Mustang Q, and ChromaSorb. The Sartobind Q and Mustang Q products contain quaternary amine ligands; whereas, ChromaSorb contains primary amine based ligands. Performance was evaluated over a range of solution conditions: 0-200 mM NaCl, pH 6.0-9.0, and flow rates of 4-20 membrane volumes/min in the presence and absence of up to 50 mM phosphate and acetate. In addition contaminant clearance was determined in the presence and absence of 5 g/L monoclonal antibody. The quaternary amine based ligands depend mainly on Coulombic interactions for removal of negatively charged contaminants. Consequently, performance of Sartobind Q and Mustang Q was compromised at high ionic strength. Primary amine based ligands in ChromaSorb enable high capacities at high ionic strength due to the presence of secondary, hydrogen bonding interactions. However, the presence of hydrogen phosphate ions leads to reduced capacity. Monoclonal antibody recovery using primary amine based anion-exchange ligands may be lower if significant binding occurs due to secondary interactions. The removal of a specific contaminant is affected by the level of removal of the other contaminants. The results of this study may be used to help guide selection of commercially available membrane absorbers for flow-through polishing steps.

  2. Ultralow protein adsorbing coatings from clickable PEG nanogel solutions: benefits of attachment under salt-induced phase separation conditions and comparison with PEG/albumin nanogel coatings.

    PubMed

    Donahoe, Casey D; Cohen, Thomas L; Li, Wenlu; Nguyen, Peter K; Fortner, John D; Mitra, Robi D; Elbert, Donald L

    2013-03-26

    Clickable nanogel solutions were synthesized by using the copper catalyzed azide/alkyne cycloaddition (CuAAC) to partially polymerize solutions of azide and alkyne functionalized poly(ethylene glycol) (PEG) monomers. Coatings were fabricated using a second click reaction: a UV thiol-yne attachment of the nanogel solutions to mercaptosilanated glass. Because the CuAAC reaction was effectively halted by the addition of a copper-chelator, we were able to prevent bulk gelation and limit the coating thickness to a single monolayer of nanogels in the absence of the solution reaction. This enabled the inclusion of kosmotropic salts, which caused the PEG to phase-separate and nearly double the nanogel packing density, as confirmed by quartz crystal microbalance with dissipation (QCM-D). Protein adsorption was analyzed by single molecule counting with total internal reflection fluorescence (TIRF) microscopy and cell adhesion assays. Coatings formed from the phase-separated clickable nanogel solutions attached with salt adsorbed significantly less fibrinogen than other 100% PEG coatings tested, as well as poly(L-lysine)-g-PEG (PLL-g-PEG) coatings. However, PEG/albumin nanogel coatings still outperformed the best 100% PEG clickable nanogel coatings. Additional surface cross-linking of the clickable nanogel coating in the presence of copper further reduced levels of fibrinogen adsorption closer to those of PEG/albumin nanogel coatings. However, this step negatively impacted long-term resistance to cell adhesion and dramatically altered the morphology of the coating by atomic force microscopy (AFM). The main benefit of the click strategy is that the partially polymerized solutions are stable almost indefinitely, allowing attachment in the phase-separated state without danger of bulk gelation, and thus producing the best performing 100% PEG coating that we have studied to date.

  3. Making Ordered DNA and Protein Structures from Computer-Printed Transparency Film Cut-Outs

    ERIC Educational Resources Information Center

    Jittivadhna, Karnyupha; Ruenwongsa, Pintip; Panijpan, Bhinyo

    2009-01-01

    Instructions are given for building physical scale models of ordered structures of B-form DNA, protein [alpha]-helix, and parallel and antiparallel protein [beta]-pleated sheets made from colored computer printouts designed for transparency film sheets. Cut-outs from these sheets are easily assembled. Conventional color coding for atoms are used…

  4. Infrared light induced patterning of proteins on ppNIPAM thermoresponsive thin films: a "protein laser printer".

    PubMed

    Cheng, Xuanhong; Yegan Erdem, E; Takeuchi, Shoji; Fujita, Hiroyuki; Ratner, Buddy D; Böhringer, Karl F

    2010-04-21

    Protein micropatterns have applications in fundamental life sciences and clinical medicine. In this work, we present a new technique to create 2-D protein micropatterns by local activation of a thin film of thermoresponsive plasma-deposited poly(N-isopropylacrylamide) (ppNIPAM) using a computer-controlled infrared laser beam. While the whole substrate is exposed to the protein solution, protein deposition happens only at laser-activated locations. A few seconds of laser exposure is all that is required to form a pattern with resolution in the single micrometre range. Successful ligand binding after protein deposition indicates that protein function remains intact after laser-induced adsorption onto ppNIPAM. This rapid, simple technique advances currently available strategies for protein patterning by its potential to pattern proteins in an enclosed environment or onto a 3-D scaffold.

  5. Properties and storage stability of whey protein edible film with spice powders.

    PubMed

    Ket-On, Apisada; Pongmongkol, Natkritta; Somwangthanaroj, Anongnat; Janjarasskul, Theeranun; Tananuwong, Kanitha

    2016-07-01

    The impact of spice powders on physical, mechanical, thermal and barrier properties, and on storage stability, of whey protein isolate (WPI)-based films was determined. Films with added spices were prepared from casting solution containing 10 % (w/w) heat-denatured WPI, glycerol (WPI:glycerol of 3:2 w/w), sodium chloride (0.4 g/100 g solution), garlic and pepper powders (≤3 g each/100 g solution). Water activity (aw) of all films was 0.53-0.57. Addition of spice powders increased thickness, darkness and yellowness of the WPI films. Films with added spices had lower tensile strength (TS), percent elongation (%E), and melting enthalpy of WPI matrices, but possessed higher water vapor permeability (WVP) than WPI film without sodium chloride and spices. The WPI film containing highest amount of garlic powder and lowest amount of pepper powder was selected for storage tests at 25-45 °C. Storage for up to 49 days resulted in reduced aw and %E, increased TS, and color changes at 35 and 45 °C, with few changes at 25 °C. However, film WVP and OP were not affected by storage conditions after 7 days storage. Active ingredients decreased over time with up to 81 % allicin and 37 % piperine retained in the film matrix after 47 days storage.

  6. Pulsed laser deposited metal oxide thin films mediated controlled adsorption of proteins

    NASA Astrophysics Data System (ADS)

    Kim, Se Jin

    Several metal oxide thin films were grown on Si substrate by pulsed laser deposition for controlling adsorption of proteins. No intentional heating of substrate and introduction of oxygen gas during growth were employed. Additionally, fibrinogen, bovine serum albumin (BSA), and lysozyme were used as model protein in this study. The film properties such as cyratllinity, surface roughness, surface electrical charge and chemistry were investigated by many techniques in order to obtain the relationship with protein adsorption. Firstly, as grown Ta2O5 and ZnO thin film were used to study the effects of surface charge on the behaviors of BSA and lysozyme adsorption. The protein thickness results by ellipsometry showed that negatively charged Ta2O5 had a stronger affinity to positively charged lysozyme, while positively charged ZnO had a stronger affinity to negatively charged BSA. The results confirmed electrostatic interaction due to surface charge is one of main factors for determining adsorption of proteins. Furthermore, annealing studies were performed by heat treatment of as grown Ta2O5 and ZnO at 800°C in air ambience. Annealed Ta2O5 thin film had almost wetting property (from 10.02° to less than 1˜2°) and the change of cystallinity (from amorphous to cyrsalline) while annealed ZnO thin film had a reduced contact angle (from 75.65° to 39.41°) and remained to crystalline structure. The fibrinogen thickness on annealed Ta2O5 film was increased compared with as grown sample, while heat treated ZnO film showed much reduction of fibrinogen adsorption. Binary Ta-Zn oxide thin films (TZ) were grown by preparing PLD target composed of 50 wt% Ta2O5 and 50 wt% ZnO. This binary film had IEP pH 7.1 indicating nearly neutral charge in pH 7.4 PBS solution, and hydrophilic property. Ellipsometrical results showed that TZ film had the lowest fibrinogen, BSA and lysozyme thickness after 120 min adsorption compared with Ta2O5 and ZnO. Other samples, bilayer oxide films in

  7. Rhodamine 6G and 800 J-heteroaggregates with enhanced acceptor luminescence (HEAL) adsorbed in transparent SiO2 GLAD thin films.

    PubMed

    Sánchez-Valencia, Juan R; Aparicio, Francisco J; Espinós, Juan P; Gonzalez-Elipe, Agustín R; Barranco, Angel

    2011-04-21

    An enhanced fluorescent emission in the near infrared is observed when the Rhodamine 800 (Rh800) and 6G (Rh6G) dyes are coadsorbed in porous SiO(2) optical thin films prepared by glancing angle deposition (GLAD). This unusual behavior is not observed in solution and it has been ascribed to the formation of a new type of J-heteroaggregates with enhanced acceptor luminescence (HEAL). This article describes in detail and explains the main features of this new phenomenology previously referred in a short communication [J. R. Sánchez-Valencia, J. Toudert, L. González-García, A. R. González-Elipe and A. Barranco, Chem. Commun., 2010, 46, 4372-4374]. It is found that the efficiency and characteristics of the energy transfer process are dependent on the Rh6G/Rh800 concentration ratio which can be easily controlled by varying the pH of the solutions used for the infiltration of the molecules or by thermal treatments. A simple model has been proposed to account for the observed enhanced acceptor luminescence in which the heteroaggregates order themselves according to a "head to tail" configuration due to the geometrical constrains imposed by the SiO(2) porous matrix thin film. The thermal stability of the dye molecules within the films and basic optical (absorption and fluorescence) principles of the HEAL process are also described.

  8. Plasticization of a protein-based film by glycerol: a spectroscopic, mechanical, and thermal study.

    PubMed

    Gao, Chunli; Stading, Mats; Wellner, Nikolaus; Parker, Mary L; Noel, Timothy R; Mills, E N Clare; Belton, Peter S

    2006-06-28

    Kafirin, the seed storage protein of the cereal sorghum, is highly homologous with the maize storage protein zein. The effects of plasticisation of a kafirin film by glycerol in the absence of water were examined by a combination of spectroscopic (NMR and infrared), rheological, and calorimetric methods. The results suggest that at low glycerol levels the glycerol is absorbed onto and possibly into the protein. Increasing the level of glycerol increases the motion of the protein and changes the protein conformation. There are corresponding changes of the mechanical properties of protein films. At 40% (w/w) of glycerol, two glass transition temperatures were observed, one of which corresponded to the glass transition temperature of pure glycerol. This result indicates that at this level of plasticizer there are sufficient glycerol/glycerol interactions occurring to allow a separate glass formation process for glycerol.

  9. Gelation Mechanisms and Characterization of Electrochemically Generated Protein Films at Metal Interfaces

    NASA Astrophysics Data System (ADS)

    Martin, Elizabeth J.

    Although the electrochemical behavior of metals used in orthopedic implants has been studied extensively, the material interactions with proteins during corrosion processes remains poorly understood. Some studies suggest that metal-protein interactions accelerate corrosion, while others suggest that proteins protect the material from degradation. Corrosion of implant materials is a major concern due to the metal ion release that can sometimes cause adverse local tissue reactions and ultimately, failure of the implant. The initial purpose of this research was therefore to study the corrosion behavior of CoCrMo, an alloy commonly used in hip replacements, with a quartz crystal microbalance (QCM) in physiologically relevant media. The QCM enables in situ characterization of surface changes accompanying corrosion and is sensitive to viscoelastic effects at its surface. Results of QCM studies in proteinaceous media showed film deposition on the alloy surface under electrochemical conditions that otherwise produced mass loss if proteins were not present in the electrolyte. Additional studies on pure Co, Cr, and Mo demonstrated that the protein films also form on Mo surfaces after a release of molybdate ions, suggesting that these ions are essential for film formation. The electrochemically generated protein films are reminiscent of carbonaceous films that form on implant surfaces in vivo, therefore a second goal of the research was to delineate mechanisms that cause the films to form. In the second stage of this research, electrochemical QCM tests were conducted on models of the CoCrMo system consisting of Cr electrodes in proteinaceous or polymeric media containing dissolved molybdate ions. Studies indicated that films can be generated through electrochemical processes so long as both amine functional groups and molybdate ions are present in the electrolyte solution. These results suggest that the films form due to an ionic cross-linking reaction between the positively

  10. Probing local water contents of in vitro protein films by ultrasonic force microscopy

    NASA Astrophysics Data System (ADS)

    Szoszkiewicz, Robert; Kulik, Andrzej J.; Gremaud, Gerard; Lekka, Malgorzata

    2005-03-01

    By means of ultrasonic force microscopy and lateral force microscopy we measure adhesion hysteresis and friction on protein films of bovine serum albumin and concanavalin A at local scales. Our investigations at different relative humidities (less than 5% and at 50% relative humidity) correspond to dehydrated and hydrated states of proteins. We demonstrate that a substantial increase of adhesion hysteresis with relative humidity is sensitive measure of protein-water binding capacity at local scales.

  11. Advances in food packaging films from milk proteins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Most commercial petroleum-based food packaging films are poor oxygen barriers, do not biodegrade, and some are suspected to even leach compounds into the food product. For instance, three-perfluorinated coatings were banned from convenience food packaging earlier this year. These shortcomings are a ...

  12. Development of ecofriendly bionanocomposite: Whey protein isolate/pullulan films with nano-SiO2.

    PubMed

    Hassannia-Kolaee, Mahbobeh; Khodaiyan, Faramarz; Pourahmad, Rezvan; Shahabi-Ghahfarrokhi, Iman

    2016-05-01

    During the past decade, the limitation of petroleum based polymers, the high price of oil, and the environmental concern were attracted the attention of researchers to develop biobased polymers. The composition of different biopolymers and the reinforcement with nano filler are common methods to improve the drawbacks of biopolymers. In this study whey protein isolate/pullulan (WPI/PUL) films contain 1%, 3%, and 5% (w/w) nano-SiO2 (NS) were prepared by a casting method. Tensile strength of nanocomposite films increased after increasing NS content, but elongation at break decreased, simultaneously. Water absorption, moisture content, solubility in water improved in the wake of increasing NS content because NS increase the cohesiveness of the polymer matrix and improved the barrier and water resistance properties of the films. water vapor permeability of film specimens decreased by increasing NS content. Uniform distribution of NS into polymer matrix was confirmed by scanning electron microscopy (SEM). XRD pattern and thermal analysis revealed increasing crystallinity and increasing Tg of film specimens with increasing NS content, respectively. According to our result WPI/PUL/NS films possess potential to be used as environment friendly packaging films to improve shelf life of food and can be used as promising alternative to petroleum based packaging films.

  13. Free-standing protein films for dynamic mode detection of cations binding

    NASA Astrophysics Data System (ADS)

    Saya, Daisuke; Coleman, Anthony W.; Lazar, Adina N.; Bergaud, Christian; Nicu, Liviu

    2005-09-01

    This letter reports on the investigation of the mechano-chemical effect of cross-linked dried free-standing alpha-lactalbumin (α-lactalbumin) thin films induced by different cation, calcium, magnesium, and potassium binding. The protein membranes were fabricated by drying droplets of an α-lactalbumin solution on top of silicon through-wafer holes obtained by deep reactive ion etching. Then the membranes were consecutively exposed to solutions of the cations in HEPES buffer solution while their resonant frequencies were measured by full-field surface stroboscopic white light interferometry. Tests on more than 30 free-standing protein films showed more significant conformational changes of the α-lactalbumin after immersion in a calcium solution than those observed after immersion in magnesium and potassium solutions. These results demonstrate, the real potential of free-standing protein films to be used as resonant biosensors for multiple cation detection.

  14. Growth of gold nanoclusters and nanocrystals induced by lysozyme protein in thin film conformation

    NASA Astrophysics Data System (ADS)

    Bhowal, Ashim Chandra; Kundu, Sarathi

    2016-08-01

    Structures and growth behavior of gold nanoclusters and nanocrystals have been explored on thin films of globular protein lysozyme by using UV-vis and photoluminescence spectroscopy, X-ray diffraction (XRD) and atomic force microscopy (AFM). A simple and one-step environment friendly method has been used to grow nanocrystals on protein surface from HAuCl4 solution. It has been found that if different interaction times are provided between lysozyme films and HAuCl4 solution, then initially formed tiny gold nanoclusters on protein surface transform into nanocrystals with the passage of time. XRD analysis shows the formation of faced-centered cubic lattice along (1 1 1) crystalline direction and AFM images confirm the presence of circular, rod-like, triangular and hexagonal crystal structures. Langmuir-like growth behavior has been identified for both the gold nanoclusters and nanocrystals formation induced by the lysozyme films, however, nanocrystal growth is relatively slower than nanocluster.

  15. Interfacial design and structure of protein/polymer films on oxidized AlGaN surfaces

    NASA Astrophysics Data System (ADS)

    Gupta, Samit K.; Wu, Hao-Hsuan; Kwak, Kwang J.; Casal, Patricia; Nicholson, Theodore R., III; Wen, Xuejin; Anisha, R.; Bhushan, Bharat; Berger, Paul R.; Lu, Wu; Brillson, Leonard J.; Lee, Stephen Craig

    2011-01-01

    Protein detection using biologically or immunologically modified field-effect transistors (bio/immunoFETs) depends on the nanoscale structure of the polymer/protein film at sensor interfaces (Bhushan 2010 Springer Handbook of Nanotechnology 3rd edn (Heidelberg: Springer); Gupta et al 2010 The effect of interface modification on bioFET sensitivity, submitted). AlGaN-based HFETs (heterojunction FETs) are attractive platforms for many protein sensing applications due to their electrical stability in high osmolarity aqueous environments and favourable current drive capabilities. However, interfacial polymer/protein films on AlGaN, though critical to HFET protein sensor function, have not yet been fully characterized. These interfacial films are typically comprised of protein-polymer films, in which analyte-specific receptors are tethered to the sensing surface with a heterobifunctional linker molecule (often a silane molecule). Here we provide insight into the structure and tribology of silane interfaces composed of one of two different silane monomers deposited on oxidized AlGaN, and other metal oxide surfaces. We demonstrate distinct morphologies and wear properties for the interfacial films, attributable to the specific chemistries of the silane monomers used in the films. For each specific silane monomer, film morphologies and wear are broadly consistent on multiple oxide surfaces. Differences in interfacial film morphology also drive improvements in sensitivity of the underlying HFET (coincident with, though not necessarily caused by, differences in interfacial film thickness). We present a testable model of the hypothetical differential interfacial depth distribution of protein analytes on FET sensor interfaces with distinct morphologies. Empirical validation of this model may rationalize the actual behaviour of planar immunoFETs, which has been shown to be contrary to expectations of bio/immunoFET behaviour prevalent in the literature for the last 20 years

  16. Novel Soluble Dietary Fiber-Tannin Self-Assembled Film: A Promising Protein Protective Material.

    PubMed

    Song, Guo-Bin; Xu, Juan; Zheng, Hua; Feng, Ying; Zhang, Wen-Wen; Li, Kun; Ge, Shuang-shuang; Li, Kai; Zhang, Hong

    2015-06-24

    In this experiment, a natural promising protein protective film was fabricated through soluble dietary fiber (SDF)-tannin nanocluster self-assembly. FT-IR, XRD, and DSC tests were employed to investigate the interaction between the SDF and tannins before and after cross-linking induced by calcium ion. On the other hand, referring to the SEM and TEM results, the self-assembly process of the protein protective film could be indicated as follows: first, calcium ion, with its cross-ability, served as the "nucleus"; SDF and tannins were combined to prepare the nanoscale SDF-tannin clusters; then, the clusters were homogeneously deposited on the surface of protein to form a protective film by self-assembling hydrogen bond between tannin component of clusters as "adhesive" and protein in aqueous solutions under very mild conditions. Film thickness could also be controlled by tannin of different concentrations ranging from 114 to 1384 μm. Antibacterial test and in vitro cytotoxicity test proved that the film had a broad spectrum of antimicrobial properties and excellent cell biocompatibility, respectively, which might open up new applications in the food preservation and biomedical fields.

  17. Ageing of solid-state protein films: Behavior of azurin at ambient conditions

    NASA Astrophysics Data System (ADS)

    Pompa, P. P.; Bramanti, A.; Maruccio, G.; Mercato, L. L. del; Cingolani, R.; Rinaldi, R.

    2005-03-01

    We report on the folding properties of the protein azurin, deposited onto SiO 2 surfaces and subsequently dehydrated. The molecular films have been maintained at ambient conditions through several days, and the ageing effects have been investigated by fluorescence spectroscopy. The experimental results show a modest initial conformational rearrangement, followed by long-term stability. Interestingly, upon rehydration of the biomolecular films at the end of the investigated period (approximately one month), azurin returns to exhibit a native-like conformation. This study indicates a rather surprising resilience of proteins to ambient conditions and sheds a somewhat unexpected positive light on reliability in biomolecular electronics.

  18. Protein Adsorption and Its Role in Bacterial Film Development

    DTIC Science & Technology

    1989-06-27

    only the secondary antibody conjugated to alkaline phosphatase was used. Combined Amino Acids as Measured by HPLC We are interested in a simple, direct...specific assay for chitin that relies on the lectin, wheat germ agglutinin (WGA). Lectins are a general class of proteins that bind to carbohydrates. The...protein; 2) a new method for measuring combined amino acids (includes proteins) in seawater was shown to measure higher concentration than the old

  19. Kinetic Effects on Self-Assembly and Function of Protein-Polymer Bioconjugates in Thin Films Prepared by Flow Coating.

    PubMed

    Chang, Dongsook; Huang, Aaron; Olsen, Bradley D

    2017-01-01

    The self-assembly of nanostructured globular protein arrays in thin films is demonstrated using protein-polymer block copolymers based on a model protein mCherry and the polymer poly(oligoethylene glycol acrylate) (POEGA). Conjugates are flow coated into thin films on a poly(ethylene oxide) grafted Si surface, forming self-assembled cylindrical nanostructures with POEGA domains selectively segregating to the air-film interface. Long-range order and preferential arrangement of parallel cylinders templated by selective surfaces are demonstrated by controlling relative humidity. Long-range order increases with coating speed when the film thicknesses are kept constant, due to reduced nucleation per unit area of drying film. Fluorescence emission spectra of mCherry in films prepared at <25% relative humidity shows a small shift suggesting that proteins are more perturbed at low humidity than high humidity or the solution state.

  20. Molecular Adsorber Coating

    NASA Technical Reports Server (NTRS)

    Straka, Sharon; Peters, Wanda; Hasegawa, Mark; Hedgeland, Randy; Petro, John; Novo-Gradac, Kevin; Wong, Alfred; Triolo, Jack; Miller, Cory

    2011-01-01

    A document discusses a zeolite-based sprayable molecular adsorber coating that has been developed to alleviate the size and weight issues of current ceramic puck-based technology, while providing a configuration that more projects can use to protect against degradation from outgassed materials within a spacecraft, particularly contamination-sensitive instruments. This coating system demonstrates five times the adsorption capacity of previously developed adsorber coating slurries. The molecular adsorber formulation was developed and refined, and a procedure for spray application was developed. Samples were spray-coated and tested for capacity, thermal optical/radiative properties, coating adhesion, and thermal cycling. Work performed during this study indicates that the molecular adsorber formulation can be applied to aluminum, stainless steel, or other metal substrates that can accept silicate-based coatings. The coating can also function as a thermal- control coating. This adsorber will dramatically reduce the mass and volume restrictions, and is less expensive than the currently used molecular adsorber puck design.

  1. Aging of whey protein films and the effect on mechanical and barrier properties.

    PubMed

    Anker, M; Stading, M; Hermansson, A M

    2001-02-01

    This work focuses on the aging of whey protein isolate (WPI) films plasticized with glycerol (G) and sorbitol (S). The films were cast from heated aqueous solutions at pH 7 and dried at 23 degrees C and 50% relative humidity (RH) for 16 h. They were stored in a climate room (23 degrees C, 50% RH) for 120 days, and the film properties were measured at regular intervals. The moisture content (MC) of the WPI/G films decreased from 22% (2 days) to 15% (45 days) and was thereafter constant at 15% (up to 120 days). This affected the mechanical properties and caused an increased stress at break (from 2.7 to 8.3 MPa), a decreased strain at break (from 33 to 4%), and an increased glass transition temperature (T(g)) (from -56 to -45 degrees C). The barrier properties were, however, unaffected, with constant water vapor permeability and a uniform film thickness. The MC of the WPI/S films was constant at approximately 9%, which gave no change in film properties.

  2. Preparation of core-shell structure Fe3 O4 @SiO2 superparamagnetic microspheres immoblized with iminodiacetic acid as immobilized metal ion affinity adsorbents for His-tag protein purification.

    PubMed

    Ni, Qian; Chen, Bing; Dong, Shaohua; Tian, Lei; Bai, Quan

    2016-04-01

    The core-shell structure Fe3 O4 /SiO2 magnetic microspheres were prepared by a sol-gel method, and immobiled with iminodiacetic acid (IDA) as metal ion affinity ligands for protein adsorption. The size, morphology, magnetic properties and surface modification of magnetic silica nanospheres were characterized by various modern analytical instruments. It was shown that the magnetic silica nanospheres exhibited superparamagnetism with saturation magnetization values of up to 58.1 emu/g. Three divalent metal ions, Cu(2+) , Ni(2+) and Zn(2+) , were chelated on the Fe3 O4 @SiO2 -IDA magnetic microspheres to adsorb lysozyme. The results indicated that Ni(2+) -chelating magnetic microspheres had the maximum adsorption capacity for lysozyme of 51.0 mg/g, adsorption equilibrium could be achieved within 60 min and the adsorbed protein could be easily eluted. Furthermore, the synthesized Fe3 O4 @SiO2 -IDA-Ni(2+) magnetic microspheres were successfully applied for selective enrichment lysozyme from egg white and His-tag recombinant Homer 1a from the inclusion extraction expressed in Escherichia coli. The result indicated that the magnetic microspheres showed unique characteristics of high selective separation behavior of protein mixture, low nonspecific adsorption, and easy handling. This demonstrates that the magnetic silica microspheres can be used efficiently in protein separation or purification and show great potential in the pretreatment of the biological sample.

  3. Properties of competitively adsorbed BSA and fibrinogen from their mixture on mixed and hybrid surfaces

    NASA Astrophysics Data System (ADS)

    Pandey, Lalit M.; Pattanayek, Sudip K.

    2013-01-01

    We have studied the adsorption of BSA and fibrinogen from their mixture onto surfaces with mixed self-assembled monolayer (SAM) of amine and octyl (ratio 1:1) and hybrid SAM. The properties of adsorbed proteins obtained from individual protein solution differ considerably from the properties of the adsorbed proteins obtained from mixture of proteins at same total concentration. The adsorbed amount of proteins is lesser and the adsorbed protein is more elastic if it is adsorbing from mixture of proteins. It is found that with increasing total protein concentration, adsorbed amount increases and elasticity of the adsorbed proteins decreases. The apparent displacements of BSA with Fb are observed on the graphs of change in frequency with time, which are obtained from quartz crystal microbalance.

  4. DNA-SMART: Biopatterned Polymer Film Microchannels for Selective Immobilization of Proteins and Cells.

    PubMed

    Schneider, Ann-Kathrin; Nikolov, Pavel M; Giselbrecht, Stefan; Niemeyer, Christof M

    2017-02-22

    A novel SMART module, dubbed "DNA-SMART" (DNA substrate modification and replication by thermoforming) is reported, where polymer films are premodified with single-stranded DNA capture strands, microthermoformed into 3D structures, and postmodified with complementary DNA-protein conjugates to realize complex biologically active surfaces within microfluidic devices. As a proof of feasibility, it is demonstrated that microchannels presenting three different proteins on their inner curvilinear surface can be used for selective capture of cells under flow conditions.

  5. Use of /γ-irradiation to produce films from whey, casein and soya proteins: structure and functionals characteristics

    NASA Astrophysics Data System (ADS)

    Lacroix, M.; Le, T. C.; Ouattara, B.; Yu, H.; Letendre, M.; Sabato, S. F.; Mateescu, M. A.; Patterson, G.

    2002-03-01

    γ-irradiation and thermal treatments have been used to produce sterilized cross-linked films. Formulations containing variable concentrations of calcium caseinate and whey proteins (whey protein isolate (WPI) and commercial whey protein concentrate) or mixture of soya protein isolate (SPI) with WPI was investigated on the physico-chemical properties of these films. Results showed that the mechanical properties of cross-linked films improved significantly the puncture strength for all types of films. Size-exclusion chromatography showed for no cross-linked proteins, a molecular mass of around 40 kDa. The soluble fractions of the cross-linked proteins molecular distributions were between 600 and 3800 kDa. γ-irradiation seems to modify to a certain extent the conformation of proteins which will adopt structures more ordered and more stable, as suggested by X-ray diffraction analysis. Microstructure observations showed that the mechanical characteristics of these films are closely related to their microscopic structure. Water vapor permeability of films based on SPI was also significantly decreased when irradiated. Microbial resistance was also evaluated for cross-linked films. Results showed that the level of biodegradation of cross-linked films was 36% after 60 d of fermentation in the presence of Pseudomonas aeruginosa.

  6. Adsorption of peptides and small proteins with control access polymer permeation to affinity binding sites. Part II: Polymer permeation-ion exchange separation adsorbents with polyethylene glycol and strong anion exchange groups.

    PubMed

    González-Ortega, Omar; Porath, Jerker; Guzmán, Roberto

    2012-03-02

    In chromatographic separations, the most general problem in small biomolecule isolation and purification is that such biomolecules are usually found in extremely low concentrations together with high concentrations of large molecular weight proteins. In the first part of this work, adsorption and size exclusion chromatography (AdSEC) controlled access media, using polyethylene glycol (PEG) as a semi-permeable barrier on a polysaccharide Immobilized Metal Affinity Chromatography (IMAC) matrix was synthesized and used to develop chromatographic adsorbents that preferentially adsorb and separate low molecular weight biomolecules while rejecting large molecular weight proteins. In this second part, we expand the concept of controlled access polymer permeation adsorption (CAPPA) media by grafting polyethylene glycol (PEG) on a high capacity polysaccharide ion exchange (IEX) chromatographic resin where PEG acts as a semi-permeable barrier that preferentially allows the permeation of small molecules while rejecting large ones. The IEX resin bearing quaternary ammonium groups binds permeated biomolecules according to their ion exchange affinity while excluding large biomolecules by the PEG barrier and thus cannot compete for the binding sites. This new AdSEC media was used to study the retention of peptides and proteins covering a wide range of molecular weights from 1 to 150 kDa. The effect of protein molecular weight towards retention by ion exchange was performed using pure protein solutions. Recovery of insulin from insulin-spiked human serum and insulin-spiked human urine was evaluated under polymer controlled permeation conditions. The CAPPA media consisted of agarose beads modified with amino-PEG-methoxy and with trimethyl ammonium groups, having chloride capacities between 20 and 40 μeq/mL and were effective in rejecting high molecular weight proteins while allowing the preferential adsorption of small proteins and peptides.

  7. Properties of a new protein film from bitter vetch (Vicia ervilia) and effect of CaCl₂ on its hydrophobicity.

    PubMed

    Arabestani, Akram; Kadivar, Mahdi; Shahedi, Mohmmad; Goli, Sayed Amir Hossein; Porta, Raffaele

    2013-06-01

    This work was aimed to investigate the potential preparation of an edible film from bitter vetch seed proteins. The film was cast from bitter vetch protein concentrate (BPC) and glycerol. CaCl₂ at the ratio of 0.1-1% (w/w) of the BPC was tested to improve film properties, specially its hydrophobicity. Some physicochemical properties of the films obtained in the absence and presence of CaCl₂ were evaluated. The results indicated that moisture content, total soluble matter, water vapour permeability and contact angle of the films prepared in the presence of CaCl₂ were significantly modified in comparison with the control values, while their mechanical properties did not significantly change. The surface morphology of the films was also considerably affected by the presence of CaCl₂. Therefore, CaCl₂ could improve BPC-films barrier properties especially their hydrophobicity, even though calcium concentration seems to be a crucial factor.

  8. In situ spectroelectrochemical and theoretical study on the oxidation of a 4H-imidazole-ruthenium dye adsorbed on nanocrystalline TiO2 thin film electrodes.

    PubMed

    Zhang, Ying; Kupfer, Stephan; Zedler, Linda; Schindler, Julian; Bocklitz, Thomas; Guthmuller, Julien; Rau, Sven; Dietzek, Benjamin

    2015-11-28

    Terpyridine 4H-imidazole-ruthenium(II) complexes are considered promising candidates for use as sensitizers in dye sensitized solar cells (DSSCs) by displaying broad absorption in the visible range, where the dominant absorption features are due to metal-to-ligand charge transfer (MLCT) transitions. The ruthenium(III) intermediates resulting from photoinduced MLCT transitions are essential intermediates in the photoredox-cycle of the DSSC. However, their photophysics is much less studied compared to the ruthenium(II) parent systems. To this end, the structural alterations accompanying one-electron oxidation of the RuIm dye series (including a non-carboxylic RuIm precursor, and, carboxylic RuImCOO in solution and anchored to a nanocrystalline TiO2 film) are investigated via in situ experimental and theoretical UV-Vis absorption and resonance Raman (RR) spectroelectrochemistry. The excellent agreement between the experimental and the TDDFT spectra derived in this work allows for an in-depth assignment of UV-Vis and RR spectral features of the dyes. A concordant pronounced wavelength dependence with respect to the charge transfer character has been observed for the model system RuIm, and both RuImCOO in solution and attached on the TiO2 surface. Excitation at long wavelengths leads to the population of ligand-to-metal charge transfer states, i.e. photoreduction of the central ruthenium(III) ion, while high-energy excitation features an intra-ligand charge transfer state localized on the 4H-imidazole moiety. Therefore, these 4H-imidazole ruthenium complexes investigated here are potential multi-photoelectron donors. One electron is donated from MLCT states, and additionally, the 4H-imidazole ligand reveals electron-donating character with a significant contribution to the excited states of the ruthenium(III) complexes upon blue-light irradiation.

  9. Kinetic Effects on Self-Assembly and Function of Protein-Polymer Bioconjugates in Thin Films Prepared by Flow Coating

    SciTech Connect

    Chang, Dongsook; Huang, Aaron; Olsen, Bradley D.

    2016-11-04

    The self-assembly of nanostructured globular protein arrays in thin films is demonstrated using protein–polymer block copolymers based on a model protein mCherry and the polymer poly(oligoethylene glycol acrylate) (POEGA). Conjugates are flow coated into thin films on a poly(ethylene oxide) grafted Si surface, forming self-assembled cylindrical nanostructures with POEGA domains selectively segregating to the air–film interface. Long-range order and preferential arrangement of parallel cylinders templated by selective surfaces are demonstrated by controlling relative humidity. Long-range order increases with coating speed when the film thicknesses are kept constant, due to reduced nucleation per unit area of drying film. Fluorescence emission spectra of mCherry in films prepared at <25% relative humidity shows a small shift suggesting that proteins are more perturbed at low humidity than high humidity or the solution state.

  10. Carbon nanodots-chitosan composite film: a platform for protein immobilization, direct electrochemistry and bioelectrocatalysis.

    PubMed

    Sheng, Meili; Gao, Yue; Sun, Junyong; Gao, Feng

    2014-08-15

    A novel composite film based on carbon nanodots (CNDs) and chitosan was readily prepared and used as immobilization matrix to entrap a heme protein, hemoglobin (Hb) for direct electrochemistry and bioelectrocatalysis. A modified electrode was obtained by casting Hb-CNDs-chitosan composites on the glassy carbon (GC) electrode surface. Spectroscopic and electrochemical studies showed that Hb entrapped in the composite film remained in its native structures, and CNDs in the film can greatly facilitate DET between the protein and the GC electrode. The electron-transfer kinetics of Hb in composite film was qualitatively evaluated by using the Marcus theory, and the apparent heterogeneous electron-transfer rate constant (ks) was estimated to be 2.39(±0.03)s(-1) with Laviron equations. The modified electrode showed excellent electrocatalytic behavior to the substrate, hydrogen peroxide (H2O2). The linear current response for H2O2 was from 1×10(-6) to 1.18×10(-4)M with a detection limit of 0.27(±0.02)μM at the signal-to-noise ratio of 3, and the apparent Michaelis-Menten constant was 0.067(±0.02)mM. These important features of CNDs-chitosan film have implied to be a promising platform for elaborating bioelectrochemical devices such as biosensors and biofuel cells.

  11. Contribution of the starch, protein, and lipid fractions to the physical, thermal, and structural properties of amaranth (Amaranthus caudatus) flour films.

    PubMed

    Tapia-Blácido, D; Mauri, A N; Menegalli, F C; Sobral, P J A; Añón, M C

    2007-06-01

    Amaranth protein-lipid (PL) and protein (P) films were elaborated and compared with amaranth flour films in order to determine the contribution of the interactions between the biopolymer (starch and protein) and the lipids to the film properties. The films were made by the casting method, using the same glycerol concentration (0.9 g glycerol/100 g solution). A separation of the lipid fraction in the PL films and a polymorphic transformation of the corresponding fatty acids were observed by differential scanning calorimetry (DSC) and verified by an analysis of the microstructure by scanning electron microscopy (SEM). The flour films showed no separation of the lipid fraction, evidence that the lipids were strongly associated with the proteins and homogenously distributed throughout the starch network, contributing to the good mechanical properties when compared to the PL films and to the excellent barrier properties when compared to both the PL and P films. The protein-protein interactions also contributed to the mechanical properties of the flour films. The presence of proteins and lipids in the flour films had an important effect on film solubility, and also on the color and opacity of the films. This study showed that the flour film properties depended on the interactions formed by their polymers (starches and proteins) and by the lipid, on the distribution of these interactions within the film matrix and on the concentrations of each component in the film.

  12. Blend-modification of soy protein/lauric acid edible films using polysaccharides.

    PubMed

    Pan, Hongyang; Jiang, Bo; Chen, Jie; Jin, Zhengyu

    2014-05-15

    Different types of polysaccharides (propyleneglycol alginate (PGA), pectin, carrageenan and aloe polysaccharide) were incorporated into soy protein isolate (SPI)/lauric acid (La) films using a co-drying process or by direct addition to form biodegradable composite films with modified water vapour permeability (WVP) and mechanical properties. The WVP of SPI/La/polysaccharide films decreased when polysaccharides were added using the co-drying process, regardless of the type of polysaccharide. The tensile strength of SPI/La film was increased by the addition of polysaccharides, and the percentage elongation at break was increased by incorporating PGA using the co-drying process. Regarding oxygen-barrier performance, no notable differences were observed between the SPI/La and SPI/La/polysaccharide films. The most significant improvement was observed by blending PGA, with the co-dried preparation exhibiting better properties than the direct-addition preparation. Scanning electron microscopy (SEM) revealed that the microstructures of the films are the basis for the differences in the barrier and mechanical properties of the modified blends of SPI, polysaccharides and La.

  13. Preparation of DNA and protein micro arrays on glass slides coated with an agarose film

    PubMed Central

    Afanassiev, Victor; Hanemann, Vera; Wölfl, Stefan

    2000-01-01

    A thin layered agarose film on microscope slides provides a versatile support for the preparation of arrayed molecular libraries. An activation step leading to the formation of aldehyde groups in the agarose creates reactive sites that allow covalent immobilization of molecules containing amino groups. Arrays of oligonucleotides and PCR products were prepared by tip printing. After hybridization with complementary fluorescence labeled nucleic acid probes strong fluorescence signals of sequence-specific binding to the immobilized probes were detected. The intensity of the fluorescence signals was proportional to the relative amount of immobilized oligonucleotides and to the concentration of the fluorescence labeled probe. We also used the agarose film-coated slides for the preparation of protein arrays. In combination with specific fluorescence labeled antibodies these protein arrays can be used for fluorescence linked immune assays. With this approach different protein tests can be performed in parallel in a single reaction with minimal amounts of the binding reagents. PMID:10871389

  14. Films.

    ERIC Educational Resources Information Center

    Philadelphia Board of Education, PA. Div. of Instructional Materials.

    The Affective Curriculum Research Project produced five films and two records during a series of experimental summer programs. The films and records form part of a curriculum designed to teach to the concerns of students. The films were an effort to describe the Philadelphia Cooperative Schools Program, to explain its importance, and to…

  15. Regenerative adsorbent heat pump

    NASA Technical Reports Server (NTRS)

    Jones, Jack A. (Inventor)

    1991-01-01

    A regenerative adsorbent heat pump process and system is provided which can regenerate a high percentage of the sensible heat of the system and at least a portion of the heat of adsorption. A series of at least four compressors containing an adsorbent is provided. A large amount of heat is transferred from compressor to compressor so that heat is regenerated. The process and system are useful for air conditioning rooms, providing room heat in the winter or for hot water heating throughout the year, and, in general, for pumping heat from a lower temperature to a higher temperature.

  16. Exploring the Properties of Genetically Engineered Silk-Elastin-Like Protein Films.

    PubMed

    Machado, Raul; da Costa, André; Sencadas, Vitor; Pereira, Ana Margarida; Collins, Tony; Rodríguez-Cabello, José Carlos; Lanceros-Méndez, Senentxu; Casal, Margarida

    2015-12-01

    Free standing films of a genetically engineered silk-elastin-like protein (SELP) were prepared using water and formic acid as solvents. Exposure to methanol-saturated air promoted the formation of aggregated β-strands rendering aqueous insolubility and improved the mechanical properties leading to a 10-fold increase in strain-to-failure. The films were optically clear with resistivity values similar to natural rubber and thermally stable up to 180 °C. Addition of glycerol showed to enhance the flexibility of SELP/glycerol films by interacting with SELP molecules through hydrogen bonding, interpenetrating between the polymer chains and granting more conformational freedom. This detailed characterization provides cues for future and unique applications using SELP based biopolymers.

  17. Fibrous parylene-C thin-film substrates for implant integration and protein assays

    NASA Astrophysics Data System (ADS)

    Wei, Lai

    Polymeric biomaterials are used in medical devices that can be surgically implanted in human beings. Long-term bio-compatibility and strong tissue integration are essential to the longevity of implanted prosthesis. Surface roughness and wettability are essential for effective cellular attachment and integration. Therefore, materials should be tailored so that their surface conditions are optimal for excellent integration with selected proteins and cells. This dissertation investigates the development of parylene-C thin films with good control over surface roughness and surface wettability. Based on these qualities, different degrees of cell and protein adhesion have been achieved, depending on surface properties and the cell/protein type. In addition, a morphology-composition gradient panel has been developed with a wide range of surface roughness and wettability, which can be used to optimize tissue growth with high-throughput screening assays and with gradient surfaces. The effects of the surface roughness and wettability of parylene-C thin films on the adhesion of human fibroblast cells and biotinylated serum proteins have been investigated. In addition, a simple method of fabricating nano-/micro-textured, free-standing, parylene-C thin-film substrate has been developed, which has been demonstrated to support cellular attachment and growth.

  18. Adsorbent and adsorbent bed for materials capture and separation processes

    DOEpatents

    Liu, Wei

    2011-01-25

    A method device and material for performing adsorption wherein a fluid mixture is passed through a channel in a structured adsorbent bed having a solid adsorbent comprised of adsorbent particles having a general diameter less than 100 um, loaded in a porous support matrix defining at least one straight flow channel. The adsorbent bed is configured to allow passage of a fluid through said channel and diffusion of a target material into said adsorbent under a pressure gradient driving force. The targeted molecular species in the fluid mixture diffuses across the porous support retaining layer, contacts the adsorbent, and adsorbs on the adsorbent, while the remaining species in the fluid mixture flows out of the channel.

  19. Molecularly imprinted protein recognition thin films constructed by controlled/living radical polymerization.

    PubMed

    Sasaki, Shogo; Ooya, Tooru; Kitayama, Yukiya; Takeuchi, Toshifumi

    2015-02-01

    We demonstrated the synthesis of molecularly imprinted polymers (MIPs) with binding affinity toward a target protein, ribonuclease A (RNase) by atom transfer radical polymerization (ATRP) of acrylic acid, acrylamide, and N,N'-methylenebisacrylamide in the presence of RNase. The binding activity of the MIPs was evaluated by surface plasmon resonance (SPR) of the MIP thin layers prepared on the gold-coated sensor chips. The MIPs prepared by ATRP (MIP-ATRP) had a binding affinity toward RNase with larger binding amount compared to MIPs prepared by conventional free radical polymerization methods (MIP-RP). Moreover, protein selectivity was evaluated using reference proteins (cytochrome c, myoglobin, and α-lactalbumin) and was confirmed in MIP-ATRP of optimum film thickness determined experimentally to be 15-30 nm; however, protein selectivity was not achieved in all MIP-RP. We have shown that ATRP is powerful technique for preparing protein recognition materials by molecular imprinting.

  20. In situ fluorescent protein imaging with metal film-enhanced total internal reflection microscopy.

    PubMed

    Burghardt, Thomas P; Charlesworth, Jon E; Halstead, Miriam F; Tarara, James E; Ajtai, Katalin

    2006-06-15

    Fluorescence detection of single molecules provides a means to investigate protein dynamics minus ambiguities introduced by ensemble averages of unsynchronized protein movement or of protein movement mimicking a local symmetry. For proteins in a biological assembly, taking advantage of the single molecule approach could require single protein isolation from within a high protein concentration milieu. Myosin cross-bridges in a muscle fiber are proteins attaining concentrations of approximately 120 muM, implying single myosin detection volume for this biological assembly is approximately 1 attoL (10(-18) L) provided that just 2% of the cross-bridges are fluorescently labeled. With total internal reflection microscopy (TIRM) an exponentially decaying electromagnetic field established on the surface of a glass-substrate/aqueous-sample interface defines a subdiffraction limit penetration depth into the sample that, when combined with confocal microscopy, permits image formation from approximately 3 attoL volumes. Demonstrated here is a variation of TIRM incorporating a nanometer scale metal film into the substrate/glass interface. Comparison of TIRM images from rhodamine-labeled cross-bridges in muscle fibers contacting simultaneously the bare glass and metal-coated interface show the metal film noticeably reduces both background fluorescence and the depth into the sample from which fluorescence is detected. High contrast metal film-enhanced TIRM images allow secondary label visualization in the muscle fibers, facilitating elucidation of Z-disk structure. Reduction of both background fluorescence and detection depth will enhance TIRM's usefulness for single molecule isolation within biological assemblies.

  1. The influence of protein aggregation on adsorption kinetics

    NASA Astrophysics Data System (ADS)

    Rovner, Joel; Roberts, Christopher; Furst, Eric; Hudson, Steven

    2015-03-01

    When proteins adsorb to an air-water interface they lower the surface tension and may form an age-dependent viscoelastic film. Protein adsorption to surfaces is relevant to both commercial uses and biological function. The rate at which the surface tension decreases depends strongly on temperature, solution pH, and protein structure. These kinetics also depend on the degree to which the protein is aggregated in solution. Here we explore these differences using Chymotrypsinogen as a model protein whose degree of aggregation is adjusted through controlled heat treatment and measured by chromatography. To study these effects we have used a micropipette tensiometer to produce a spherical-cap bubble whose interfacial pressure was controlled - either steady or oscillating. Short heat treatment produced small soluble aggregates, and these adsorbed faster than the original protein monomer. Longer heat treatment produced somewhat larger soluble aggregates which adsorbed more slowly. These results point to complex interactions during protein adsorption.

  2. Improvement in physical and biological properties of chitosan/soy protein films by surface grafted heparin.

    PubMed

    Wang, Xiaomei; Hu, Ling; Li, Chen; Gan, Li; He, Meng; He, Xiaohua; Tian, Weiqun; Li, Mingming; Xu, Li; Li, Yinping; Chen, Yun

    2016-02-01

    A series of chitosan/soy protein isolate (SPI) composite films (CS-n, n=0, 10 and 30, corresponding to SPI content in the composites) were prepared. Heparin was grafted onto the surface of CS-n to fabricate a series of heparinized films (HCS-n). CS-n and HCS-n were characterized by ATR-Fourier transform infrared spectroscopy and water contact angle. The surface heparin density was measured by toluidine blue assay. The results showed that heparin has been successfully grafted onto the surface of CS-n. Heparin evenly distributed on the surface of the films and the heparin content increased with the increase of SPI content, and the hydrophilicity of the films was enhanced due to the grafted heparin. The cytocompatibility and hemocompatibility of CS-n and HCS-n were evaluated by cell culture (MTT assay, live/dead assay, cell morphology and cell density observation), platelet adhesion test, plasma recalcification time (PRT) measurement, hemolysis assay and thrombus formation test. HCS-n showed higher cell adhesion rate and improved cytocompatibility compared to the corresponding CS-n. HCS-n also exhibited lower platelet adhesion, longer PRT, higher blood anticoagulant indexes (BCI) and lower hemolysis rate than the corresponding CS-n. The improved cytocompatibility and hemocompatibility of HCS-n would shed light on the potential applications of chitosan/soy protein-based biomaterials that may come into contact with blood.

  3. Real-time, step-wise, electrical detection of protein molecules using dielectrophoretically aligned SWNT-film FET aptasensors.

    PubMed

    An, Taechang; Kim, Ki Su; Hahn, Sei Kwang; Lim, Geunbae

    2010-08-21

    Aptamer functionalized addressable SWNT-film arrays between cantilever electrodes were successfully developed for biosensor applications. Dielectrophoretically aligned SWNT suspended films made possible highly specific and rapid detection of target proteins with a large binding surface area. Thrombin aptamer immobilized SWNT-film FET biosensor resulted in a real-time, label-free, and electrical detection of thrombin molecules down to a concentration of ca. 7 pM with a step-wise rapid response time of several seconds.

  4. The characterization of protein release from sericin film in the presence of an enzyme: towards fibroblast growth factor-2 delivery.

    PubMed

    Nishida, Ayumu; Naganuma, Tsuyoshi; Kanazawa, Takanori; Takashima, Yuuki; Yamada, Masaki; Okada, Hiroaki

    2011-07-29

    Aqueous preparations of silk protein (sericin) films were prepared to evaluate their biodegradation properties. In the absence of trypsin, sericin film swelled rapidly, kept its shape, and remained unaltered for 28 days or longer due to form β-sheet structures. In the presence of trypsin, sericin film gradually degraded; since the rate depended on the concentration of trypsin, the films likely underwent enzymatic hydrolysis. Sericin film incorporating the model protein drug fluorescein isothiocyanate-albumin (FA) also gradually degraded in the presence of trypsin and resulted in the sustained release of FA for 2 weeks or longer; in contrast, FA release was quite slow in the absence of trypsin. It is expected that sericin film has potential as a biodegradable and drug-releasing carrier. To evaluate the practical applicability of sericin film for the repair of defective tissues, fibroblast growth factor-2 (FGF-2) was incorporated into sericin films and the films were implanted on skull defects in rats. Whereas FGF-2 release was suppressed in the absence of trypsin in vitro, it appears that FGF-2, immobilized by ionic interactions between sericin and FGF-2, can be sustained-released in vivo from films incorporating 2500 or 250 ng of FGF-2 to support the growth of tissue around wounds.

  5. Combinatorial Synthesis of and high-throughput protein release from polymer film and nanoparticle libraries.

    PubMed

    Petersen, Latrisha K; Chavez-Santoscoy, Ana V; Narasimhan, Balaji

    2012-09-06

    Polyanhydrides are a class of biomaterials with excellent biocompatibility and drug delivery capabilities. While they have been studied extensively with conventional one-sample-at-a-time synthesis techniques, a more recent high-throughput approach has been developed enabling the synthesis and testing of large libraries of polyanhydrides(1). This will facilitate more efficient optimization and design process of these biomaterials for drug and vaccine delivery applications. The method in this work describes the combinatorial synthesis of biodegradable polyanhydride film and nanoparticle libraries and the high-throughput detection of protein release from these libraries. In this robotically operated method (Figure 1), linear actuators and syringe pumps are controlled by LabVIEW, which enables a hands-free automated protocol, eliminating user error. Furthermore, this method enables the rapid fabrication of micro-scale polymer libraries, reducing the batch size while resulting in the creation of multivariant polymer systems. This combinatorial approach to polymer synthesis facilitates the synthesis of up to 15 different polymers in an equivalent amount of time it would take to synthesize one polymer conventionally. In addition, the combinatorial polymer library can be fabricated into blank or protein-loaded geometries including films or nanoparticles upon dissolution of the polymer library in a solvent and precipitation into a non-solvent (for nanoparticles) or by vacuum drying (for films). Upon loading a fluorochrome-conjugated protein into the polymer libraries, protein release kinetics can be assessed at high-throughput using a fluorescence-based detection method (Figures 2 and 3) as described previously(1). This combinatorial platform has been validated with conventional methods(2) and the polyanhydride film and nanoparticle libraries have been characterized with (1)H NMR and FTIR. The libraries have been screened for protein release kinetics, stability and

  6. Hybrid polymer-lipid films as platforms for directed membrane protein insertion.

    PubMed

    Kowal, Justyna; Wu, Dalin; Mikhalevich, Viktoria; Palivan, Cornelia G; Meier, Wolfgang

    2015-05-05

    Hybrids composed of amphiphilic block copolymers and lipids constitute a new generation of biological membrane-inspired materials. Hybrid membranes resulting from self-assembly of lipids and polymers represent adjustable models for interactions between artificial and natural membranes, which are of key importance, e.g., when developing systems for drug delivery. By combining poly(dimethylsiloxane)-block-poly(2-methyl-2-oxazoline) amphiphilic copolymers (PDMS-b-PMOXA) with various phospholipids, we obtained hybrid films with modulated properties and topology, based on phase separation, and the formation of distinct domains. By understanding the factors driving the phase separation in these hybrid lipid-polymer films, we were able to use them as platforms for directed insertion of membrane proteins. Tuning the composition of the polymer-lipids mixtures favored successful insertion of membrane proteins with desired topological distributions (in polymer or/and lipid regions). Controlled insertion and location of membrane proteins in hybrid films make these hybrids ideal candidates for numerous applications where specific spatial functionality is required.

  7. Relationship between the microstructure and the mechanical and barrier properties of whey protein films.

    PubMed

    Anker, M; Stading, M; Hermansson, A M

    2000-09-01

    This work was focused on the relationship between the microstructure and the mechanical and barrier properties of whey protein isolate (WPI) films. Sorbitol (S) and glycerol (G) were used as plasticizers and the pH was varied between 7 and 9. The films were cast from heated aqueous solutions and dried in a climate room at 23 degrees C and 50% relative humidity for 16 h. The microstructure of the films was found to be dependent on the concentration, the plasticizers, and the pH. When the concentration increased, a more aggregated structure was formed, with a denser protein network and larger pores. This resulted in increased water vapor permeability (WVP) and decreased oxygen permeability (OP). When G was used as a plasticizer instead of S, the microstructure was different, and the moisture content and WVP approximately doubled. When the pH increased from 7 to 9, a denser protein structure was formed, the strain at break increased, and the OP decreased.

  8. Amaranth proteins foaming properties: Film rheology and foam stability - Part 2.

    PubMed

    Bolontrade, Agustín J; Scilingo, Adriana A; Añón, María C

    2016-05-01

    In this work the influence of pH and ionic strength on the stability of foams prepared with amaranth protein isolate was analyzed. The behaviour observed was related to the physico-chemical and structural changes undergone by amaranth protein as a result of those treatments. The results obtained show that foams prepared at acidic pH were more stable than the corresponding to alkaline pH. At pH 2.0 the foams presented higher times and more volumes of drainage. This behaviour is consistent with the characteristics of the interfacial film, which showed a higher viscoelasticity and a greater flexibility at acidic pH than alkaline pH value, which in turn increased by increasing the concentration of proteins in the foaming solution. It is also important to note that the presence of insoluble protein is not necessarily detrimental to the properties of the foam. Detected changes in the characteristics of the interfacial film as in the foam stability have been attributed to the increased unfolding, greater flexibility and net charge of amaranth proteins at acidic conditions.

  9. Corneal Cell Adhesion to Contact Lens Hydrogel Materials Enhanced via Tear Film Protein Deposition

    PubMed Central

    Elkins, Claire M.; Qi, Qin M.; Fuller, Gerald G.

    2014-01-01

    Tear film protein deposition on contact lens hydrogels has been well characterized from the perspective of bacterial adhesion and viability. However, the effect of protein deposition on lens interactions with the corneal epithelium remains largely unexplored. The current study employs a live cell rheometer to quantify human corneal epithelial cell adhesion to soft contact lenses fouled with the tear film protein lysozyme. PureVision balafilcon A and AirOptix lotrafilcon B lenses were soaked for five days in either phosphate buffered saline (PBS), borate buffered saline (BBS), or Sensitive Eyes Plus Saline Solution (Sensitive Eyes), either pure or in the presence of lysozyme. Treated contact lenses were then contacted to a live monolayer of corneal epithelial cells for two hours, after which the contact lens was sheared laterally. The apparent cell monolayer relaxation modulus was then used to quantify the extent of cell adhesion to the contact lens surface. For both lens types, lysozyme increased corneal cell adhesion to the contact lens, with the apparent cell monolayer relaxation modulus increasing up to an order of magnitude in the presence of protein. The magnitude of this increase depended on the identity of the soaking solution: lenses soaked in borate-buffered solutions (BBS, Sensitive Eyes) exhibited a much greater increase in cell attachment upon protein addition than those soaked in PBS. Significantly, all measurements were conducted while subjecting the cells to moderate surface pressures and shear rates, similar to those experienced by corneal cells in vivo. PMID:25144576

  10. Corneal cell adhesion to contact lens hydrogel materials enhanced via tear film protein deposition.

    PubMed

    Elkins, Claire M; Qi, Qin M; Fuller, Gerald G

    2014-01-01

    Tear film protein deposition on contact lens hydrogels has been well characterized from the perspective of bacterial adhesion and viability. However, the effect of protein deposition on lens interactions with the corneal epithelium remains largely unexplored. The current study employs a live cell rheometer to quantify human corneal epithelial cell adhesion to soft contact lenses fouled with the tear film protein lysozyme. PureVision balafilcon A and AirOptix lotrafilcon B lenses were soaked for five days in either phosphate buffered saline (PBS), borate buffered saline (BBS), or Sensitive Eyes Plus Saline Solution (Sensitive Eyes), either pure or in the presence of lysozyme. Treated contact lenses were then contacted to a live monolayer of corneal epithelial cells for two hours, after which the contact lens was sheared laterally. The apparent cell monolayer relaxation modulus was then used to quantify the extent of cell adhesion to the contact lens surface. For both lens types, lysozyme increased corneal cell adhesion to the contact lens, with the apparent cell monolayer relaxation modulus increasing up to an order of magnitude in the presence of protein. The magnitude of this increase depended on the identity of the soaking solution: lenses soaked in borate-buffered solutions (BBS, Sensitive Eyes) exhibited a much greater increase in cell attachment upon protein addition than those soaked in PBS. Significantly, all measurements were conducted while subjecting the cells to moderate surface pressures and shear rates, similar to those experienced by corneal cells in vivo.

  11. Films

    NASA Astrophysics Data System (ADS)

    Li, Ming; Zhang, Yang; Shao, Yayun; Zeng, Min; Zhang, Zhang; Gao, Xingsen; Lu, Xubing; Liu, J.-M.; Ishiwara, Hiroshi

    2014-09-01

    In this paper, we investigated the microstructure and electrical properties of Bi2SiO5 (BSO) doped SrBi2Ta2O9 (SBT) films deposited by chemical solution deposition. X-ray diffraction observation indicated that the crystalline structures of all the BSO-doped SBT films are nearly the same as those of a pure SBT film. Through BSO doping, the 2Pr and 2Ec values of SBT films were changed from 15.3 μC/cm2 and 138 kV/cm of pure SBT to 1.45 μC/cm2 and 74 kV/cm of 10 wt.% BSO-doped SBT. The dielectric constant at 1 MHz for SBT varied from 199 of pure SBT to 96 of 10 wt.% BSO-doped SBT. The doped SBT films exhibited higher leakage current than that of non-doped SBT films. Nevertheless, all the doped SBT films still had small dielectric loss and low leakage current. Our present work will provide useful insights into the BSO doping effects to the SBT films, and it will be helpful for the material design in the future nonvolatile ferroelectric memories.

  12. Lubrication of metal-on-metal hip joints: the effect of protein content and load on film formation and wear.

    PubMed

    Myant, C; Underwood, R; Fan, J; Cann, P M

    2012-02-01

    Lubricant films were measured for a series of bovine serum and protein containing (albumin, globulin) saline solutions for CoCrMo femoral component sliding against a glass disc. Central film thickness was measured by optical interferometry as a function of time (constant mean speed: 0 and 10 mm/s) and variable mean speed (0-50 mm/s). The effect of load (5-20 N) on film thickness was also studied. The development of the wear scar on the CoCrMo surface was monitored by measuring the width of the contact zone during the film thickness tests. The results showed film thickness increased with time for both the static and sliding tests. Films formed in the static, loaded test were typically in the range of 3-40 nm. The globulin containing solutions formed the thickest films. In the sliding tests a wear scar rapidly formed on the implant component for the bovine serum and albumin fluids, negligible wear was observed for the globulin solutions. Film thickness increased with sliding time for all test solutions and was much greater than predicted by isoviscous EHL models. The film increase was found to correlate with increasing wear scar size and thus decreasing contact pressure. A new lubricating mechanism is proposed whereby during sliding the fluid undergoes bulk phase separation rheology, so that an elevated protein phase forms in the inlet zone. This protein phase is a high-viscosity biphasic matrix, which is periodically entrained into the contact forming a thick protective hydro-gel film. One of the main findings of this study is that film thickness was very sensitive to load; to a much greater extent than predicted by EHL models. Thus film formation in MoM hip joints is very susceptible to high contact pressures which might be due to implant misalignment and edge-loading.

  13. Compositional and physicochemical factors governing the viability of Lactobacillus rhamnosus GG embedded in starch-protein based edible films

    PubMed Central

    Soukoulis, Christos; Singh, Poonam; Macnaughtan, William; Parmenter, Christopher; Fisk, Ian D.

    2016-01-01

    Probiotic incorporation in edible films and coatings has been shown recently to be an efficient strategy for the delivery of probiotics in foods. In the present work, the impact of the compositional, physicochemical and structural properties of binary starch-protein edible films on Lactobacillus rhamnosus GG viability and stability was evaluated. Native rice and corn starch, as well as bovine skin gelatine, sodium caseinate and soy protein concentrate were used for the fabrication of the probiotic edible films. Starch and protein type both impacted the structural, mechanical, optical and thermal properties of the films, and the process loss of L. rhamnosus GG during evaporation-dehydration was significantly lower in the presence of proteins (0.91–1.07 log CFU/g) compared to solely starch based systems (1.71 log CFU/g). A synergistic action between rice starch and proteins was detected when monitoring the viability of L. rhamnosus GG over four weeks at fridge and room temperature conditions. In particular, a 3- to 7-fold increase in the viability of L. rhamnosus GG was observed in the presence of proteins, with sodium caseinate – rice starch based films offering the most enhanced stability. The film's shelf-life (as calculated using the FAO/WHO (2011) basis of 6 log viable CFU/g) ranged between 27-96 and 15–24 days for systems stored at fridge or room temperature conditions respectively. PMID:26726280

  14. 'All-solid-state' electrochemistry of a protein-confined polymer electrolyte film

    SciTech Connect

    Parthasarathy, Meera; Pillai, Vijayamohanan K. Mulla, Imtiaz S.; Shabab, Mohammed; Khan, M.I.

    2007-12-07

    Interfacial redox behavior of a heme protein (hemoglobin) confined in a solid polymer electrolyte membrane, Nafion (a perfluoro sulfonic acid ionomer) is investigated using a unique 'all-solid-state' electrochemical methodology. The supple phase-separated structure of the polymer electrolyte membrane, with hydrophilic pools containing solvated protons and water molecules, is found to preserve the incorporated protein in its active form even in the solid-state, using UV-visible, Fluorescence (of Tryptophan and Tyrosine residues) and DRIFT (diffuse reflectance infrared Fourier transform) spectroscopy. More specifically, solid-state cyclic voltammetry and electrochemical impedance of the protein-incorporated polymer films reveal that the Fe{sup 2+}-form of the entrapped protein is found to bind molecular oxygen more strongly than the native protein. In the 'all-solid-state' methodology, as there is no need to dip the protein-modified electrode in a liquid electrolyte (like the conventional electrochemical methods), it offers an easier means to study a number of proteins in a variety of polymer matrices (even biomimetic assemblies). In addition, the results of the present investigation could find interesting application in a variety of research disciplines, in addition to its fundamental scientific interest, including protein biotechnology, pharmaceutical and biomimetic chemistry.

  15. Novel adhesive properties of poly(ethylene-oxide) adsorbed nanolayers

    NASA Astrophysics Data System (ADS)

    Zeng, Wenduo

    Solid-polymer interfaces play crucial roles in the multidisciplinary field of nanotechnology and are the confluence of physics, chemistry, biology, and engineering. There is now growing evidence that polymer chains irreversibly adsorb even onto weakly attractive solid surfaces, forming a nanometer-thick adsorbed polymer layer ("adsorbed polymer nanolayers"). It has also been reported that the adsorbed layers greatly impact on local structures and properties of supported polymer thin films. In this thesis, I aim to clarify adhesive and tribological properties of adsorbed poly(ethylene-oxide) (PEO) nanolayers onto silicon (Si) substrates, which remain unsolved so far. The adsorbed nanolayers were prepared by the established protocol: one has to equilibrate the melt or dense solution against a solid surface; the unadsorbed chains can be then removed by a good solvent, while the adsorbed chains are assumed to maintain the same conformation due to the irreversible freezing through many physical solid-segment contacts. I firstly characterized the formation process and the surface/film structures of the adsorbed nanolayers by using X-ray reflectivity, grazing incidence X-ray diffraction, and atomic force microscopy. Secondly, to compare the surface energy of the adsorbed layers with the bulk, static contact angle measurements with two liquids (water and glycerol) were carried out using a optical contact angle meter equipped with a video camera. Thirdly, I designed and constructed a custom-built adhesion-testing device to quantify the adhesive property. The experimental results provide new insight into the microscopic structure - macroscopic property relationship at the solid-polymer interface.

  16. Processing of mussel-adhesive protein analog copolymer thin films by matrix-assisted pulsed laser evaporation

    NASA Astrophysics Data System (ADS)

    Patz, T.; Cristescu, R.; Narayan, R.; Menegazzo, N.; Mizaikoff, B.; Messersmith, P. B.; Stamatin, I.; Mihailescu, I. N.; Chrisey, D. B.

    2005-07-01

    We have demonstrated the successful thin film growth of a mussel-adhesive protein analog, DOPA-modified PEO-PPO-PEO block copolymer PF127, using matrix-assisted pulsed laser evaporation (MAPLE). The MAPLE-deposited thin films were examined using Fourier transform infrared spectroscopy, atomic force microscopy, X-ray photoelectron spectroscopy, and contact-angle measurements. We have found that the main functional groups of the mussel-adhesive protein analog are present in the transferred film. These adhesive materials have several potential electronic, medical, and marine applications.

  17. Study of water vapour permeability of protein and gum-based edible films by a photothermal method

    NASA Astrophysics Data System (ADS)

    Tomás, S. A.; Saavedra, R.; Cruz, A.; Pedroza-Islas, R.; San Martín, E.

    2005-06-01

    The water vapour permeability of protein and gum-based edible films was studied by means of a photothermal method. The films were prepared with two basic ingredients, whey protein concentrate and mesquite gum, according to the proportions 75:25, 50:50, 25:75, and 0:100 (weight:weight). The water vapour diffusion coefficient of the analyzed films was found within the interval 0.37 × 10-6 to 2.04 × 10-6 cm^2/s, increasing linearly by increasing the mesquite gum composition in the films. The incorporation of mesquite gum in films produces less effective moisture barriers due to its highly hydrophilic property.

  18. Chemical treatment and characterization of soybean straw and soybean protein isolate/straw composite films.

    PubMed

    Martelli-Tosi, Milena; Assis, Odílio B G; Silva, Natália C; Esposto, Bruno S; Martins, Maria Alice; Tapia-Blácido, Delia R

    2017-02-10

    This work investigated changes in the chemical composition and structure of soybean straw (SS) treated with alkali (NaOH 5% and 17.5%) and bleached with hydrogen peroxide (H2O2) or sodium hypochlorite (NaOCl). Removal of the amorphous constituents increased the degree of crystallinity and the content of cellulose fibers particularly after reaction with high concentrations of alkali. Treatment with NaOH 17.5% contributed to the allomorph transition from cellulose I to II regardless of the bleaching agent, but H2O2 as bleaching agent promoted more effective delignification. This work also evaluated the potential use of treated and non-treated SS as reinforcement filler in soy protein isolate film (SPI). Films added with treated SS presented higher mechanical resistance, lower elongation at break, and lower solubility in water. Addition of non-treated SS did not affect the properties of the SPI film significantly. The low solubility and the reasonable water vapor permeability of the composite films make them suitable packaging materials for fresh fruit and vegetables.

  19. Properties of whey protein isolate nanocomposite films reinforced with nanocellulose isolated from oat husk.

    PubMed

    Qazanfarzadeh, Zeinab; Kadivar, Mahdi

    2016-10-01

    Whey protein isolate (WPI)-based composite films with varying proportions of oat husk nanocellulose (ONC) obtained from acid sulfuric hydrolysis were prepared using a solution casting method. The obtained material after each step of the isolating cellulose, morphological and crystallinity of the ONC were studied by Fourier transform infrared (FTIR) spectroscopy, Scanning electron microscopy (SEM) and X-ray diffraction (XRD), respectively. The effect of ONC content (0, 2.5, 5 and 7.5wt% of WPI) on physical, mechanical and barrier properties of the nanocomposite were then evaluated. FTIR spectroscopy indicated the progressive removal of non-cellulosic components from the oat husk. SEM images showed the mean width of ONC was about 76nm and XRD analysis revealed the crystallinity increased after acid hydrolysis. The films prepared with up to 5wt% ONC showed the highest tensile strength, Young's modulus, solubility and the lowest elongation at break and moisture content. At high level (7.5wt%), tensile strength, Young's modulus and solubility of the films decreased and elongation at break and moisture content increased due to agglomeration of the fillers. Nevertheless, film transparency and water vapor permeability decreased with ONC incorporation.

  20. Protein-modified shear mode film bulk acoustic resonator for bio-sensing applications

    NASA Astrophysics Data System (ADS)

    Wang, Jingjing; Liu, Weihui; Xu, Yan; Chen, Da; Li, Dehua; Zhang, Luyin

    2014-09-01

    In this paper, we present a shear mode film bulk acoustic biosensor based on micro-electromechanical technology. The film bulk acoustic biosensor is a diaphragmatic structure consisting of a lateral field excited ZnO piezoelectric film piezoelectric stack built on an Si3N4 membrane. The device works at near 1.6 GHz with Q factors of 579 in water and 428 in glycerol. A frequency shift of 5.4 MHz and a small decline in the amplitude are found for the measurements in glycerol compared with those in water because of the viscous damping derived from the adjacent glycerol. For bio-sensing demonstration, the resonator was modified with biotin molecule to detect protein-ligand interactions in real-time and in situ. The resonant frequency of the biotin-modified device drops rapidly and gradually reaches equilibrium when exposed to the streptavidin solution due to the biotin-streptavidin interaction. The proposed film bulk acoustic biosensor shows promising applications for disease diagnostics, prognosis, and drug discovery.

  1. Soluble Proteins Form Film by the Treatment of Low Temperature Plasma

    NASA Astrophysics Data System (ADS)

    Ikehara, Sanae; Sakakita, Hajime; Ishikawa, Kenji; Akimoto, Yoshihiro; Nakanishi, Hayao; Shimizu, Nobuyuki; Hori, Masaru; Ikehara, Yuzuru

    2015-09-01

    It has been pointed out that low temperature plasma in atmosphere was feasible to use for hemostasis without heat injury. Indeed, earlier studies demonstrated that low temperature plasma played an important role to stimulate platelets to aggregate and turned on the proteolytic activities of coagulation factors, resulting in the acceleration of the natural blood coagulation process. On the other hands, our developed equips could immediately form clots upon the contact with plasma flair, while the histological appearance was different from natural coagulation. Based on these findings in formed clots, we sought to determine if plasma flair supplied by our devices was capable of forming film using a series of soluble proteins Following plasma treatment, films were formed from bovine serum albumin, and the other plasma proteins at physiological concentration. Analysis of trans-electron microscope demonstrated that plasma treatment generated small protein particles and made them fuse to be larger aggregations The combined results demonstrated that plasma are capable of aggregating soluble proteins and that platelets and coagulation factors are not necessary for plasma induced blood coagulation. Supported in part by Grants-in-Aid for Scientific Research on Priority Area (21590454, 24590498, and 24108006 to Y. I.).

  2. Moisture Sensitivity, Optical, Mechanical and Structural Properties of Whey Protein-Based Edible Films Incorporated with Rapeseed Oil

    PubMed Central

    Kadzińska, Justyna

    2016-01-01

    Summary The objective of this work is to study the effect of the rapeseed oil content on the physical properties of whey protein emulsion films. For this purpose, whey protein films with the addition of 0, 1, 2 and 3% of rapeseed oil, and glycerol as a plasticizer were obtained by the casting method. Film-forming emulsions were evaluated and compared using light scattering granulometry. The Sauter mean diameters (d32) of lipid droplets in film-forming solutions showed an increasing trend when increasing the oil volume fractions. The inclusion of rapeseed oil enhanced the hydrophobic character of whey protein films, reducing moisture content and film solubility in water. All emulsified films showed high lightness (L*≈90). Parameter a* decreased and parameter b* and total colour difference (∆E) increased with the increase of the volume fractions of oil. These results were consistent with visual observations; control films were transparent and those containing oil opaque. Water vapour sorption experimental data at the full range of water activity values from 0.11 to 0.93 were well described with Peleg’s equation (R2≥0.99). The tensile strength, Young’s modulus and elongation at break increased with the increase of rapeseed oil volume fraction, which could be explained by interactions between lipids and the protein matrix. These results revealed that rapeseed oil has enormous potential to be incorporated into whey protein to make edible film or coating for some food products. The mechanical resistance decreased with the addition of the lipids, and the opacity and soluble matter content increased. PMID:27904396

  3. Moisture Sensitivity, Optical, Mechanical and Structural Properties of Whey Protein-Based Edible Films Incorporated with Rapeseed Oil.

    PubMed

    Galus, Sabina; Kadzińska, Justyna

    2016-03-01

    The objective of this work is to study the effect of the rapeseed oil content on the physical properties of whey protein emulsion films. For this purpose, whey protein films with the addition of 0, 1, 2 and 3% of rapeseed oil, and glycerol as a plasticizer were obtained by the casting method. Film-forming emulsions were evaluated and compared using light scattering granulometry. The Sauter mean diameters (d32) of lipid droplets in film-forming solutions showed an increasing trend when increasing the oil volume fractions. The inclusion of rapeseed oil enhanced the hydrophobic character of whey protein films, reducing moisture content and film solubility in water. All emulsified films showed high lightness (L*≈90). Parameter a* decreased and parameter b* and total colour difference (∆E) increased with the increase of the volume fractions of oil. These results were consistent with visual observations; control films were transparent and those containing oil opaque. Water vapour sorption experimental data at the full range of water activity values from 0.11 to 0.93 were well described with Peleg's equation (R(2)≥0.99). The tensile strength, Young's modulus and elongation at break increased with the increase of rapeseed oil volume fraction, which could be explained by interactions between lipids and the protein matrix. These results revealed that rapeseed oil has enormous potential to be incorporated into whey protein to make edible film or coating for some food products. The mechanical resistance decreased with the addition of the lipids, and the opacity and soluble matter content increased.

  4. Extracellular matrix proteins as temporary coating for thin-film neural implants

    NASA Astrophysics Data System (ADS)

    Ceyssens, Frederik; Deprez, Marjolijn; Turner, Neill; Kil, Dries; van Kuyck, Kris; Welkenhuysen, Marleen; Nuttin, Bart; Badylak, Stephen; Puers, Robert

    2017-02-01

    Objective. This study investigates the suitability of a thin sheet of extracellular matrix (ECM) proteins as a resorbable coating for temporarily reinforcing fragile or ultra-low stiffness thin-film neural implants to be placed on the brain, i.e. microelectrocorticographic (µECOG) implants. Approach. Thin-film polyimide-based electrode arrays were fabricated using lithographic methods. ECM was harvested from porcine tissue by a decellularization method and coated around the arrays. Mechanical tests and an in vivo experiment on rats were conducted, followed by a histological tissue study combined with a statistical equivalence test (confidence interval approach, 0.05 significance level) to compare the test group with an uncoated control group. Main results. After 3 months, no significant damage was found based on GFAP and NeuN staining of the relevant brain areas. Significance. The study shows that ECM sheets are a suitable temporary coating for thin µECOG neural implants.

  5. Antimicrobial activity of biodegradable polysaccharide and protein-based films containing active agents.

    PubMed

    Kuorwel, Kuorwel K; Cran, Marlene J; Sonneveld, Kees; Miltz, Joseph; Bigger, Stephen W

    2011-04-01

    Significant interest has emerged in the introduction of food packaging materials manufactured from biodegradable polymers that have the potential to reduce the environmental impacts associated with conventional packaging materials. Current technologies in active packaging enable effective antimicrobial (AM) packaging films to be prepared from biodegradable materials that have been modified and/or blended with different compatible materials and/or plasticisers. A wide range of AM films prepared from modified biodegradable materials have the potential to be used for packaging of various food products. This review examines biodegradable polymers derived from polysaccharides and protein-based materials for their potential use in packaging systems designed for the protection of food products from microbial contamination. A comprehensive table that systematically analyses and categorizes much of the current literature in this area is included in the review.

  6. Protein film voltammetry and co-factor electron transfer dynamics in spinach photosystem II core complex.

    PubMed

    Zhang, Yun; Magdaong, Nikki; Frank, Harry A; Rusling, James F

    2014-05-01

    Direct protein film voltammetry (PFV) was used to investigate the redox properties of the photosystem II (PSII) core complex from spinach. The complex was isolated using an improved protocol not used previously for PFV. The PSII core complex had high oxygen-evolving capacity and was incorporated into thin lipid and polyion films. Three well-defined reversible pairs of reduction and oxidation voltammetry peaks were observed at 4 °C in the dark. Results were similar in both types of films, indicating that the environment of the PSII-bound cofactors was not influenced by film type. Based on comparison with various control samples including Mn-depleted PSII, peaks were assigned to chlorophyll a (Chl a) (Em = -0.47 V, all vs. NHE, at pH 6), quinones (-0.12 V), and the manganese (Mn) cluster (Em = 0.18 V). PFV of purified iron heme protein cytochrome b-559 (Cyt b-559), a component of PSII, gave a partly reversible peak pair at 0.004 V that did not have a potential similar to any peaks observed from the intact PSII core complex. The closest peak in PSII to 0.004 V is the 0.18 V peak that was found to be associated with a two-electron process, and thus is inconsistent with iron heme protein voltammetry. The -0.47 V peak had a peak potential and peak potential-pH dependence similar to that found for purified Chl a incorporated into DMPC films. The midpoint potentials reported here may differ to various extents from previously reported redox titration data due to the influence of electrode double-layer effects. Heterogeneous electron transfer (hET) rate constants were estimated by theoretical fitting and digital simulations for the -0.47 and 0.18 V peaks. Data for the Chl a peaks were best fit to a one-electron model, while the peak assigned to the Mn cluster was best fit by a two-electron/one-proton model.

  7. Resistance to protein adsorption and adhesion of fibroblasts on nanocrystalline diamond films: the role of topography and boron doping.

    PubMed

    Alcaide, María; Papaioannou, Stavros; Taylor, Andrew; Fekete, Ladislav; Gurevich, Leonid; Zachar, Vladimir; Pennisi, Cristian Pablo

    2016-05-01

    Boron-doped nanocrystalline diamond (BNCD) films exhibit outstanding electrochemical properties that make them very attractive for the fabrication of electrodes for novel neural interfaces and prosthetics. In these devices, the physicochemical properties of the electrode materials are critical to ensure an efficient long-term performance. The aim of this study was to investigate the relative contribution of topography and doping to the biological performance of BNCD films. For this purpose, undoped and boron-doped NCD films were deposited on low roughness (LR) and high roughness (HR) substrates, which were studied in vitro by means of protein adsorption and fibroblast growth assays. Our results show that BNCD films significantly reduce the adsorption of serum proteins, mostly on the LR substrates. As compared to fibroblasts cultured on LR BNCD films, cells grown on the HR BNCD films showed significantly reduced adhesion and lower growth rates. The mean length of fibronectin fibrils deposited by the cells was significantly increased in the BNCD coated substrates, mainly in the LR surfaces. Overall, the largest influence on protein adsorption, cell adhesion, proliferation, and fibronectin deposition was due to the underlying sub-micron topography, with little or no influence of boron doping. In perspective, BNCD films displaying surface roughness in the submicron range may be used as a strategy to reduce the fibroblast growth on the surface of neural electrodes.

  8. Resistance to moist conditions of whey protein isolate and pea starch biodegradable films and low density polyethylene nondegradable films: a comparative study

    NASA Astrophysics Data System (ADS)

    Mehyar, G. F.; Bawab, A. Al

    2015-10-01

    Biodegradable packaging materials are degraded under the natural environmental conditions. Therefore using them could alleviate the problem of plastics accumulation in nature. For effective replacement of plastics, with biodegradable materials, biodegradable packages should keep their properties under the high relative humidity (RH) conditions. Therefore the objectives of the study were to develop biodegradable packaging material based on whey protein isolate (WPI) and pea starch (PS). To study their mechanical, oxygen barrier and solubility properties under different RHs compared with those of low density polyethylene (LDPE), the most used plastic in packaging. Films of WPI and PS were prepared separately and conditioned at different RH (30-90%) then their properties were studied. At low RHs (<50%), WPI films had 2-3 times lower elongation at break (E or stretchability) than PS and LDPE. Increasing RH to 90% significantly (P<0.01) increased the elongation of PS but not WPI and LDPE films. LDPE and WPI films kept significantly (P<0.01) higher tensile strength (TS) than PS films at high RH (90%). Oxygen permeability (OP) of all films was very low (<0.5 cm3 μm m-2 d-1 kPa-1) below 40% RH but increased for PS films and became significantly (P<0.01) different than that of LDPE and WPI at > 40% RH. Oxygen permeability of WPI and LDPE did not adversely affected by increasing RH to 65%. Furthermore, WPI and LDPE films had lower degree of hydration at 50% and 90% RH and total soluble matter than PS films. These results suggest that WPI could be successfully replacing LDPE in packaging of moist products.

  9. Phosphorylated, cellulose-based substrates as potential adsorbents for bone morphogenetic proteins in biomedical applications: a protein adsorption screening study using cytochrome C as a bone morphogenetic protein mimic.

    PubMed

    Mucalo, Michael R; Kato, Katsuya; Yokogawa, Yoshiyuki

    2009-06-01

    Screening studies aimed at identifying useful biomedical materials that (when combined with implants) can attract bone morphogenetic proteins to their surfaces have been conducted. In this paper, the screening process has involved carrying out protein adsorption studies using cytochrome C, as a BMP protein mimic on phosphorylated cellulose-based substrates. These studies have shown that phosphorylation of cellulose produces materials that are capable of attracting the adsorption of cytochrome C to their surface. In contrast, negligible cytochrome C adsorption was observed on the unphosphorylated cellulose-based materials. The selective uptake of the positively charged cytochrome C (from solutions at pH 9.51) by the negatively charged phosphorylated cotton and microcrystalline cellulose substrates was primarily due to this protein's high isoelectric point (i.e.p) of 9.8 which gives it a positive charge at pHprotein adsorption behaviour, this property with respect to phosphorylated materials and its potential use for selective BMP adsorption onto biomedical materials, have not been reported directly in the literature. The work thus shows that the phosphorylated cellulose-based substrates should be seriously considered as carrier materials that could be used (with preloaded BMPs) as part of an implant system to assist in implant healing.

  10. Enhanced physicochemical properties of chitosan/whey protein isolate composite film by sodium laurate-modified TiO2 nanoparticles.

    PubMed

    Zhang, Wei; Chen, Jiwang; Chen, Yue; Xia, Wenshui; Xiong, Youling L; Wang, Hongxun

    2016-03-15

    Chitosan/whey protein isolate film incorporated with sodium laurate-modified TiO2 nanoparticles was developed. The nanocomposite film was characterized by scanning electron microscopy, X-ray diffraction and differential scanning calorimetry, and investigated in physicochemical properties as color, tensile strength, elongation at break, water vapor permeability and water adsorption isotherm. Our results showed that the nanoparticles improved the compatibility of whey protein isolate and chitosan. Addition of nanoparticles increased the whiteness of chitosan/whey protein isolate film, but decreased its transparency. Compared with binary film, the tensile strength and elongation at break of nanocomposite film were increased by 11.51% and 12.01%, respectively, and water vapor permeability was decreased by 7.60%. The equilibrium moisture of nanocomposite film was lower than binary film, and its water sorption isotherm of the nanocomposite film fitted well to Guggenheim-Anderson-deBoer model. The findings contributed to the development of novel food packaging materials.

  11. The improvement of characteristics of biodegradable films made from kefiran-whey protein by nanoparticle incorporation.

    PubMed

    Zolfi, Mohsen; Khodaiyan, Faramarz; Mousavi, Mohammad; Hashemi, Maryam

    2014-08-30

    Biodegradable kefiran-whey protein isolate (WPI) nanocomposites were produced using montmorillonite (MMT) and nano-TiO2 as nanoparticles in the percentage of 1, 3, and 5% (w/w) by a casting and solvent-evaporation method. Physical, mechanical, and water-vapor permeability (WVP) properties were determined as a function of nanoparticle concentration. The results revealed that the effect of these nanoparticles was different according to their nature and percentage. The films incorporated with 5% (w/w) MMT showed the highest tensile strength, Young's modulus, puncture strength, and the lowest WVP compared with the control and TiO2 added films. In contrast to MMT, addition of TiO2 nanoparticles due to the plasticizing effect led to a significant change in color and transparency of nanocomposite. Scanning electron microscopy (SEM) observations demonstrated the films' properties in relation to their microstructures. The surface topography results also showed a considerable increase in roughness parameters by incorporating the nanoparticles in kefiran-WPI matrix.

  12. Layer-by-Layer Deposition with Polymers Containing Nitrilotriacetate, A Convenient Route to Fabricate Metal- and Protein-Binding Films.

    PubMed

    Wijeratne, Salinda; Liu, Weijing; Dong, Jinlan; Ning, Wenjing; Ratnayake, Nishanka Dilini; Walker, Kevin D; Bruening, Merlin L

    2016-04-27

    This paper describes a convenient synthesis of nitrilotriacetate (NTA)-containing polymers and subsequent layer-by-layer adsorption of these polymers on flat surfaces and in membrane pores. The resulting films form NTA-metal-ion complexes and capture 2-3 mmol of metal ions per mL of film. Moreover, these coatings bind multilayers of polyhistidine-tagged proteins through association with NTA-metal-ion complexes. Inclusion of acrylic acid repeat units in NTA-containing copolymers promotes swelling to increase protein binding in films on Au-coated wafers. Adsorption of NTA-containing films in porous nylon membranes gives materials that capture ∼46 mg of His-tagged ubiquitin per mL. However, the binding capacity decreases with the protein molecular weight. Due to the high affinity of NTA for metal ions, the modified membranes show modest leaching of Ni(2+) in binding and rinsing buffers. Adsorption of NTA-containing polymers is a simple method to create metal- and protein-binding films and may, with future enhancement of stability, facilitate development of disposable membranes that rapidly purify tagged proteins.

  13. Effect of replacement of corn starch by whey protein isolate in biodegradable film blends obtained by extrusion.

    PubMed

    Azevedo, Viviane Machado; Borges, Soraia Vilela; Marconcini, José Manoel; Yoshida, Maria Irene; Neto, Alfredo Rodrigues Sena; Pereira, Tamara Coelho; Pereira, Camila Ferreira Gonçalves

    2017-02-10

    The aim of this study was to evaluate the effect of replacing corn starch by whey protein isolated (WPI) in biodegradable polymer blends developed by extrusion. X-ray diffraction showed the presence of a Vh-type crystalline arrangement. The films were homogeneous, indicating strong interfacial adhesion between the protein and the thermoplastic starch matrix (TPS) as observed in scanning electron microscopy. The addition of WPI on TPS matrix promoted an increase in the thermal stability of the materials. It was observed 58.5% decrease in the water vapor permeability. The effect of corn starch substitution by WPI on mechanical properties resulted in a more resistant and less flexible film when compared the TPS film. The addition of WPI caused greenish yellow color and less transparent films. The substitution of corn starch by WPI made it possible to obtain polymer blends with improved properties and represents an innovation for application as a packaging material.

  14. Protein-Containing Lipid Bilayers Intercalated with Size-Matched Mesoporous Silica Thin Films.

    PubMed

    Isaksson, Simon; Watkins, Erik B; Browning, Kathryn L; Kjellerup Lind, Tania; Cárdenas, Marité; Hedfalk, Kristina; Höök, Fredrik; Andersson, Martin

    2017-01-11

    Proteins are key components in a multitude of biological processes, of which the functions carried out by transmembrane (membrane-spanning) proteins are especially demanding for investigations. This is because this class of protein needs to be incorporated into a lipid bilayer representing its native environment, and in addition, many experimental conditions also require a solid support for stabilization and analytical purposes. The solid support substrate may, however, limit the protein functionality due to protein-material interactions and a lack of physical space. We have in this work tailored the pore size and pore ordering of a mesoporous silica thin film to match the native cell-membrane arrangement of the transmembrane protein human aquaporin 4 (hAQP4). Using neutron reflectivity (NR), we provide evidence of how substrate pores host the bulky water-soluble domain of hAQP4, which is shown to extend 7.2 nm into the pores of the substrate. Complementary surface analytical tools, including quartz crystal microbalance with dissipation monitoring (QCM-D) and fluorescence microscopy, revealed successful protein-containing supported lipid bilayer (pSLB) formation on mesoporous silica substrates, whereas pSLB formation was hampered on nonporous silica. Additionally, electron microscopy (TEM and SEM), light scattering (DLS and stopped-flow), and small-angle X-ray scattering (SAXS) were employed to provide a comprehensive characterization of this novel hybrid organic-inorganic interface, the tailoring of which is likely to be generally applicable to improve the function and stability of a broad range of membrane proteins containing water-soluble domains.

  15. Electrochemical hydrogenation of a homogeneous nickel complex to form a surface adsorbed hydrogen-evolving species.

    PubMed

    Martin, Daniel J; McCarthy, Brian D; Donley, Carrie L; Dempsey, Jillian L

    2015-03-28

    A Ni(II) complex degrades electrochemically in the presence of acid in acetonitrile to form an electrode adsorbed film that catalytically evolves hydrogen. Comparison with a similar compound permitted investigation of the degradation mechanism.

  16. Nanoporous membrane based on block copolymer thin film for protein drug delivery

    NASA Astrophysics Data System (ADS)

    Yang, Seung Yun; Yang, Jeong-A.; Kim, Eung-Sam; Jeon, Gumhye; Oh, Eun Ju; Choi, Kwan Yong; Hahn, Sei Kwang; Kim, Jin Kon

    2010-03-01

    We studied long term and controlled release of protein drugs by using nanoporous membranes with various pore sizes. Nanoporous membrane consists of the separation layer prepared by polystyrene-block-poly(methylmethacrylate) copolymer thin film and conventional microfiltration membrane as a support. We demonstrate a long-term constant in vitro release of bovine serum albumin (BSA)and human growth hormone ) (hGH) without their denaturation up to 2 months. A nearly constant serum concentration of hGH was maintained up to 3 weeks in SD rats. The long-term constant delivery based on this membrane for protein drugs within the therapeutic range can be highly appreciated for the patients with hormone- deficiency.

  17. One step physically adsorbed coating of silica capillary with excellent stability for the separation of basic proteins by capillary zone electrophoresis.

    PubMed

    Guo, Xiao-Feng; Guo, Xiao-Mei; Wang, Hong; Zhang, Hua-Shan

    2015-11-01

    The coating of capillary inner surface is considered to be an effective approach to suppress the adsorption of proteins on capillary inner surface in CE. However, most of coating materials reported are water-soluble, which may dissolve in BGE during the procedure of electrophoresis. In this study, a novel strategy for selection of physically coating materials has been illustrated to get coating layer with excellent stability using materials having poor solubility in commonly used solvents. Taking natural chitin as example (not hydrolyzed water soluble chitosan), a simple one step coating method using chitin solution in hexafluoroisopropanol was adopted within only 21 min with good coating reproducibility (RSDs of EOF for within-batch coated capillaries of 1.55% and between-batch coated capillaries of 2.31%), and a separation of four basic proteins on a chitin coated capillary was performed to evaluate the coating efficacy. Using chitin coating, the adsorption of proteins on capillary inner surface was successfully suppressed with reversed and stable EOF, and four basic proteins including lysozyme, cytochrome c, ribonuclease A and α-chymotrypsinogen A were baseline separated within 16 min with satisfied separation efficiency using 20 mM pH 2.0 H3PO4-Na2HPO4 as back ground electrolyte and 20 kV as separation voltage. What is more important, the chitin coating layer could be stable for more than two months during this study, which demonstrates that chitin is an ideal material for preparing semi-permanent coating on bare fused silica capillary inner wall and has hopeful potential in routine separation of proteins with CE.

  18. Epichlorohydrin-Cross-linked Hydroxyethyl Cellulose/Soy Protein Isolate Composite Films as Biocompatible and Biodegradable Implants for Tissue Engineering.

    PubMed

    Zhao, Yanteng; He, Meng; Zhao, Lei; Wang, Shiqun; Li, Yinping; Gan, Li; Li, Mingming; Xu, Li; Chang, Peter R; Anderson, Debbie P; Chen, Yun

    2016-02-03

    A series of epichlorohydrin-cross-linked hydroxyethyl cellulose/soy protein isolate composite films (EHSF) was fabricated from hydroxyethyl cellulose (HEC) and soy protein isolate (SPI) using a process involving blending, cross-linking, solution casting, and evaporation. The films were characterized with FTIR, solid-state (13)C NMR, UV-vis spectroscopy, and mechanical testing. The results indicated that cross-linking interactions occurred in the inter- and intramolecules of HEC and SPI during the fabrication process. The EHSF films exhibited homogeneous structure and relative high light transmittance, indicating there was a certain degree of miscibility between HEC and SPI. The EHSF films exhibited a relative high mechanical strength in humid state and an adjustable water uptake ratio and moisture absorption ratio. Cytocompatibility, hemocompatibility and biodegradability were evaluated by a series of in vitro and in vivo experiments. These results showed that the EHSF films had good biocompatibility, hemocompatibility, and anticoagulant effect. Furthermore, EHSF films could be degraded in vitro and in vivo, and the degradation rate could be controlled by adjusting the SPI content. Hence, EHSF films might have a great potential for use in the biomedical field.

  19. Surface field of forces and protein adsorption behavior of poly(hydroxyethylmethacrylate) films deposited from plasma.

    PubMed

    Morra, M; Cassinelli, C

    1995-01-01

    Polymeric films were deposited from hydroxyethylmethacrylate (HEMA) plasma on non-woven poly(butyleneterephtalate) (PBT) filter materials. To test the effect of deposition conditions on surface properties, film were deposited using a constant monomer flow rate and a discharge power ranging from 40-100 W. Surface composition and surface energetics were evaluated by Electron Spectroscopy for Chemical Analysis (ESCA) and contact angle measurement, respectively. Albumin (Alb) and fibrinogen (Fg) adsorption from single protein solutions to the plasma-coated filters was measured. Results illustrate the marked effects of the deposition condition on the surface composition, the surface field of forces, and the protein adsorption behavior. The latter is modeled by the application of the Good-van Oss-Chaudhury theory of Lewis acid-base contribution to interfacial energetics. Materials endowed with widely different properties are obtained from the same monomer and different deposition conditions, a result that must be taken into account both in the production step, to assure constant quality, and in the development of specifically tailored materials.

  20. Development of Eco-friendly Soy Protein Isolate Films with High Mechanical Properties through HNTs, PVA, and PTGE Synergism Effect

    NASA Astrophysics Data System (ADS)

    Liu, Xiaorong; Song, Ruyuan; Zhang, Wei; Qi, Chusheng; Zhang, Shifeng; Li, Jianzhang

    2017-03-01

    This study was to develop novel soy protein isolate-based films for packaging using halloysite nanotubes (HNTs), poly-vinyl alcohol (PVA), and 1,2,3-propanetriol-diglycidyl-ether (PTGE). The structural, crystallinity, opacity, micromorphology, and thermal stability of the resultant SPI/HNTs/PVA/PTGE film were analyzed by the Attenuated total reflectance-Fourier transformed infrared (ATR-FTIR) spectroscopy, X-ray diffraction (XRD), UV-Vis spectrophotometry, scanning electron microscopy (SEM), and thermo-gravimetric analysis (TGA). The SPI/HNTs/PVA/PTGE film illustrated that HNTs were uniformly dispersed in the SPI matrix and the thermal stability of the film was enhanced. Furthermore, the tensile strength (TS) of the SPI/HNTs/PVA/PTGE film was increased by 329.3% and the elongation at the break (EB) remained unchanged. The water absorption (WA) and the moisture content (MC) were decreased by 5.1% and 10.4%, respectively, compared to the unmodified film. The results highlighted the synergistic effects of SPI, HNTs, PVA, and PTGE on the mechanical properties, water resistance, and thermal stability of SPI films, which showed excellent strength and flexibility. In short, SPI films prepared from HNTs, PVA, and PTGE showed considerable potential as packaging materials.

  1. Development of Eco-friendly Soy Protein Isolate Films with High Mechanical Properties through HNTs, PVA, and PTGE Synergism Effect

    PubMed Central

    Liu, Xiaorong; Song, Ruyuan; Zhang, Wei; Qi, Chusheng; Zhang, Shifeng; Li, Jianzhang

    2017-01-01

    This study was to develop novel soy protein isolate-based films for packaging using halloysite nanotubes (HNTs), poly-vinyl alcohol (PVA), and 1,2,3-propanetriol-diglycidyl-ether (PTGE). The structural, crystallinity, opacity, micromorphology, and thermal stability of the resultant SPI/HNTs/PVA/PTGE film were analyzed by the Attenuated total reflectance-Fourier transformed infrared (ATR-FTIR) spectroscopy, X-ray diffraction (XRD), UV-Vis spectrophotometry, scanning electron microscopy (SEM), and thermo-gravimetric analysis (TGA). The SPI/HNTs/PVA/PTGE film illustrated that HNTs were uniformly dispersed in the SPI matrix and the thermal stability of the film was enhanced. Furthermore, the tensile strength (TS) of the SPI/HNTs/PVA/PTGE film was increased by 329.3% and the elongation at the break (EB) remained unchanged. The water absorption (WA) and the moisture content (MC) were decreased by 5.1% and 10.4%, respectively, compared to the unmodified film. The results highlighted the synergistic effects of SPI, HNTs, PVA, and PTGE on the mechanical properties, water resistance, and thermal stability of SPI films, which showed excellent strength and flexibility. In short, SPI films prepared from HNTs, PVA, and PTGE showed considerable potential as packaging materials. PMID:28281634

  2. Development of Eco-friendly Soy Protein Isolate Films with High Mechanical Properties through HNTs, PVA, and PTGE Synergism Effect.

    PubMed

    Liu, Xiaorong; Song, Ruyuan; Zhang, Wei; Qi, Chusheng; Zhang, Shifeng; Li, Jianzhang

    2017-03-10

    This study was to develop novel soy protein isolate-based films for packaging using halloysite nanotubes (HNTs), poly-vinyl alcohol (PVA), and 1,2,3-propanetriol-diglycidyl-ether (PTGE). The structural, crystallinity, opacity, micromorphology, and thermal stability of the resultant SPI/HNTs/PVA/PTGE film were analyzed by the Attenuated total reflectance-Fourier transformed infrared (ATR-FTIR) spectroscopy, X-ray diffraction (XRD), UV-Vis spectrophotometry, scanning electron microscopy (SEM), and thermo-gravimetric analysis (TGA). The SPI/HNTs/PVA/PTGE film illustrated that HNTs were uniformly dispersed in the SPI matrix and the thermal stability of the film was enhanced. Furthermore, the tensile strength (TS) of the SPI/HNTs/PVA/PTGE film was increased by 329.3% and the elongation at the break (EB) remained unchanged. The water absorption (WA) and the moisture content (MC) were decreased by 5.1% and 10.4%, respectively, compared to the unmodified film. The results highlighted the synergistic effects of SPI, HNTs, PVA, and PTGE on the mechanical properties, water resistance, and thermal stability of SPI films, which showed excellent strength and flexibility. In short, SPI films prepared from HNTs, PVA, and PTGE showed considerable potential as packaging materials.

  3. Mechanical and water barrier properties of isolated soy protein composite edible films as affected by carvacrol and cinnamaldehyde micro and nanoemulsions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Edible films may be used in food packaging, for which they must deliver good barrier and mechanical properties. Films based on proteins have good gas barrier and mechanical properties, but poor water barrier properties. Films made from lipids have good water barrier properties, but poor mechanical p...

  4. Layer-by-layer assembly of bi-protein/layered double hydroxide ultrathin film and its electrocatalytic behavior for catechol.

    PubMed

    Kong, Xianggui; Rao, Xiuying; Han, Jingbin; Wei, Min; Duan, Xue

    2010-10-15

    This paper reports the fabrication of a bi-protein/layered double hydroxide (LDH) ultrathin film in which hemoglobin (HB) and horseradish peroxidase (HRP) molecules were assembled alternately with LDH nanosheets via the layer-by-layer (LBL) deposition technique, and its electrocatalytic performances for oxidation of catechol were demonstrated. The results of XRD indicate that the HB-HRP/LDH ultrathin film possesses a long range stacking order in the normal direction of the substrate, with the two proteins accommodated in the LDH gallery respectively as monolayer arrangement. SEM images show that the film surface exhibits a continuous and uniform morphology, and AFM reveals the Root-Mean-Square (RMS) roughness of ∼10.2 nm for the film. A stable direct electrochemical redox behavior of the proteins was successfully obtained for the HB-HRP/LDH film modified electrode. In addition, it exhibits remarkable electrocatalytic activity towards oxidation of catechol, based on the synergistic effect of the two proteins. The catechol biosensor in this work displays a wide linear response range (6-170 μM, r=0.999), low detection limit (5 μM), high sensitivity and good reproducibility.

  5. Investigations on the Q and CT Bands of Cytochrome c Submonolayer Adsorbed on an Alumina Surface Using Broadband Spectroscopy with Single-Mode Integrated Optical Waveguides.

    PubMed

    Wiederkehr, Rodrigo S; Hoops, Geoffrey C; Aslan, Mustafa M; Byard, Courtney L; Mendes, Sergio B

    2009-05-14

    In this work, we report experimental results on the molar absorptivity of cytochrome c adsorbed at different submonolayer levels onto an aluminum oxide waveguide surface; our data show a clear dependence of the protein optical properties on its surface density. The measurements were performed using the broadband, single-mode, integrated optical waveguide spectroscopic technique, which is an extremely sensitive tool able to reach submonolayer levels of detection required for this type of studies. This investigation focuses on the molar absorptivity at the Q-band (centered at 525 nm) and, for the first time to our knowledge, the weak charge transfer (CT) band (centered at 695 nm) of surface-adsorbed cyt c. Polarized light in the spectral region from 450 to 775 nm was all-coupled into an alumina thin film, which functioned as a single-mode planar optical waveguide. The alumina thin-film waveguide used for this work had a thickness of 180 nm and was deposited on a glass substrate by the atomic layer deposition process. The protein submonolayer was formed on the alumina waveguide surface through electrostatic adsorption from an aqueous buffer solution at neutral pH. The optical properties of the surface-adsorbed cyt c were investigated for bulk protein concentrations ranging from 5 nM to 8200 nM in the aqueous buffer solution. For a protein surface density of 2.3 pmol/cm(2), the molar absorptivity measured at the charge transfer band was 335 M(-1) cm(-1), and for a surface density of 15 pmol/cm(2) was 720 M(-1) cm(-1), which is much closer to the value of cyt c dissolved in an aqueous neutral buffer (830 M(-1) cm(-1)). The modification of the protein molar absorptivity and its dependence on the surface density can most likely be attributed to conformational changes of the surface-adsorbed species.

  6. Conformation and topology of amyloid beta-protein adsorbed on a tethered artificial membrane probed by surface plasmon field-enhanced fluorescence spectroscopy.

    PubMed

    Song, Haipeng; Ritz, Sandra; Knoll, Wolfgang; Sinner, Eva-Kathrin

    2009-10-01

    Progressive depositions of cerebral amyloid are primary neuropathologic features of Alzheimer's disease (AD). The amyloid is composed of a 39-42 amino acid peptide called the amyloid beta-protein (Abeta). Repeated investigation suggests that the conformational transition of Abeta from alpha-helix or random coil to beta-sheet structure plays a key role in the inappropriate accumulation of cerebral amyloid plaques. In this manuscript, we describe a fluorescence-based immunoassay technology to investigate the conformation and topology of Abeta peptides interacting with peptide-tethered planar lipid bilayers. Dual monoclonal antibodies (mAbs) labelled with fluorophores were employed to recognise a linear N- and a beta-sheet C-terminus of Abeta peptides on the model membrane, respectively. Kinetics of antibody-Abeta binding were determined by surface plasmon field-enhanced fluorescence spectroscopy (SPFS). The conformational transition of Abeta by melatonin, a defined beta-sheet breaker, was probed using paired monoclonal antibodies. The Abeta interaction with the membrane was evaluated by carefully analyzing the change in kinetic/affinity parameters in the presence or absence of melatonin. These results show that SPFS can be used to examine conformational transition of Abeta on an artificial membrane, providing a novel and versatile platform for conveniently monitoring protein-membrane interaction and screening for new beta-sheet breakers.

  7. Antioxidant activities of distiller dried grains with solubles as protein films containing tea extracts and their application in the packaging of pork meat.

    PubMed

    Yang, Hyun-Ju; Lee, Ji-Hyeon; Won, Misun; Song, Kyung Bin

    2016-04-01

    Distiller dried grains with solubles (DDGS) as protein (DP) films were prepared. Additionally, to prepare anti-oxidant films, green tea extract (GTE), oolong tea extract (OTE), and black tea extract (BTE) were incorporated into the DP films. Consequently, the incorporation of the tea extracts did not alter the physical properties of the films much, whereas the antioxidant activities, such as ABTS and DPPH radical scavenging activities were observed. To apply the DP films containing tea extracts to food packaging, pork meat was wrapped with the films and stored at 4 °C for 10 d. During storage, the pork meat wrapped with the DP films containing GTE, OTE, and BTE had less lipid oxidation than did the control. Among the tea extracts, the DP film containing GTE had the greatest antioxidant activity. These results indicate that the DP films containing green tea extracts can be utilized as an anti-oxidative packaging material for pork meat.

  8. Preparation and characterization of soy protein films with a durable water resistance-adjustable and antimicrobial surface.

    PubMed

    Li, Shuzhao; Donner, Elizabeth; Xiao, Huining; Thompson, Michael; Zhang, Yachuan; Rempel, Curtis; Liu, Qiang

    2016-12-01

    A water resistant surface was first obtained by immobilizing hydrophobic copolymers, poly (styrene-co-glycidyl methacrylate) (PSG), with functional groups on soy protein isolate (SPI) films. XPS and AFM results showed that PSG copolymers were immobilized on the film by chemical bonding, and formed a rough surface with some bumps because of the segregation of two different phases on PSG copolymers. Water resistance of the modified films could be adjusted dramatically by further immobilizing different amounts of guanidine-based antimicrobial polymers, poly (hexamethylene guanidine hydrochloride) (PHMG) on the resulting hydrophobic surface. The introduction of hydrophilic PHMG on the resulting surface generated many micropores, which potentially increased the water uptake of the modified films. Furthermore, the modified SPI films showed higher thermostability compared to native SPI film and broad-spectrum antimicrobial activity by contact killing, attributed to the presence of PHMG on the surface. The modified SPI film with a multi-functional surface showed potential for applications in the packaging and medical fields.

  9. Controllable degradation of medical magnesium by electrodeposited composite films of mussel adhesive protein (Mefp-1) and chitosan.

    PubMed

    Jiang, Ping-Li; Hou, Rui-Qing; Chen, Cheng-Dong; Sun, Lan; Dong, Shi-Gang; Pan, Jin-Shan; Lin, Chang-Jian

    2016-09-15

    To control the degradation rate of medical magnesium in body fluid environment, biocompatible films composed of Mussel Adhesive Protein (Mefp-1) and chitosan were electrodeposited on magnesium surface in cathodic constant current mode. The compositions and structures of the films were characterized by atomic force microscope (AFM), scanning electron microscope (SEM) and infrared reflection absorption spectroscopy (IRAS). And the corrosion protection performance was investigated using electrochemical measurements and immersion tests in simulated body fluid (Hanks' solution). The results revealed that Mefp-1 and chitosan successfully adhered on the magnesium surface and formed a protective film. Compared with either single Mefp-1 or single chitosan film, the composite film of chitosan/Mefp-1/chitosan (CPC (chitosan/Mefp-1/chitosan)) exhibited lower corrosion current density, higher polarization resistance and more homogenous corrosion morphology and thus was able to effectively control the degradation rate of magnesium in simulated body environment. In addition, the active attachment and spreading of MC3T3-E1 cells on the CPC film coated magnesium indicated that the CPC film was significantly able to improve the biocompatibility of the medical magnesium.

  10. Enhancement of Colorimetric Response of Enzymatic Reactions by Thermally Evaporated Plasmonic Thin Films: Application to Glial Fibrillary Acidic Protein.

    PubMed

    Abel, Biebele; Kabir, Tabassum S; Odukoya, Babatunde; Mohammed, Muzaffer; Aslan, Kadir

    2015-02-07

    We report the enhancement of the colorimetric response of horseradish peroxidase (HRP) and alkaline phosphatase (AP) in bioassays by thermally evaporated silver, gold, copper and nickel thin films. In this regard, a model bioassay based on biotin-avidin interactions was employed. Biotin groups and enzymes were introduced to all surfaces using a biotinylated linker molecule and avidin, respectively. The colorimetric response of HRP in the model bioassay carried out on the plasmonic thin films were up to 4.4-fold larger as compared to control samples (i.e., no plasmonic thin films), where the largest enhancement of colorimetric response was observed on silver thin films. The colorimetric response of AP on plasmonic thin films was found to be similar to those observed on control samples, which was attributed to the loss of enzymes from the surface during the bioassay steps. The extent of enzymes immobilized on to plasmonic thin films was found to affect the colorimetric response of the model bioassay. These findings allowed us to demonstrate the use of silver thin films for the detection of glial fibrillary acidic protein (GFAP), where the colorimetric response of the standard bioassays for GFAP was enhanced up to 67% as compared to bioassays on glass slides.

  11. Enhancement of Colorimetric Response of Enzymatic Reactions by Thermally Evaporated Plasmonic Thin Films: Application to Glial Fibrillary Acidic Protein

    PubMed Central

    Abel, Biebele; Kabir, Tabassum S.; Odukoya, Babatunde; Mohammed, Muzaffer; Aslan, Kadir

    2015-01-01

    We report the enhancement of the colorimetric response of horseradish peroxidase (HRP) and alkaline phosphatase (AP) in bioassays by thermally evaporated silver, gold, copper and nickel thin films. In this regard, a model bioassay based on biotin-avidin interactions was employed. Biotin groups and enzymes were introduced to all surfaces using a biotinylated linker molecule and avidin, respectively. The colorimetric response of HRP in the model bioassay carried out on the plasmonic thin films were up to 4.4-fold larger as compared to control samples (i.e., no plasmonic thin films), where the largest enhancement of colorimetric response was observed on silver thin films. The colorimetric response of AP on plasmonic thin films was found to be similar to those observed on control samples, which was attributed to the loss of enzymes from the surface during the bioassay steps. The extent of enzymes immobilized on to plasmonic thin films was found to affect the colorimetric response of the model bioassay. These findings allowed us to demonstrate the use of silver thin films for the detection of glial fibrillary acidic protein (GFAP), where the colorimetric response of the standard bioassays for GFAP was enhanced up to 67% as compared to bioassays on glass slides. PMID:25663850

  12. Film thickness dependence of protein adsorption from blood serum and plasma onto poly(sulfobetaine)-grafted surfaces.

    PubMed

    Yang, Wei; Chen, Shengfu; Cheng, Gang; Vaisocherová, Hana; Xue, Hong; Li, Wei; Zhang, Jinli; Jiang, Shaoyi

    2008-09-02

    In this work, we investigate protein adsorption from single protein solutions and complex media such as 100% blood serum and plasma onto poly(sulfobetaine methacrylate) (polySBMA)-grafted surfaces via atom transfer radical polymerization (ATRP) at varying film thicknesses. It is interesting to observe that protein adsorption exhibits a minimum at a medium film thickness. Results show that the surface with 62 nm polySBMA brushes presents the best nonfouling character in 100% blood serum and plasma although all of these surfaces are highly resistant to nonspecific protein adsorption from single fibrinogen and lysozyme solutions. Surface resistance to 100% blood serum or plasma is necessary for many applications from blood-contacting devices to drug delivery. This work provides a new in vitro evaluation standard for the application of biomaterials in vivo.

  13. MTBE adsorption on alternative adsorbents and packed bed adsorber performance.

    PubMed

    Rossner, Alfred; Knappe, Detlef R U

    2008-04-01

    Widespread use of the fuel additive methyl tertiary-butyl ether (MTBE) has led to frequent MTBE detections in North American and European drinking water sources. The overall objective of this research was to evaluate the effectiveness of a silicalite zeolite, a carbonaceous resin, and a coconut-shell-based granular activated carbon (GAC) for the removal of MTBE from water. Isotherm and short bed adsorber tests were conducted in ultrapure water and river water to obtain parameters describing MTBE adsorption equilibria and kinetics and to quantify the effect of natural organic matter (NOM) on MTBE adsorption. Both the silicalite zeolite and the carbonaceous resin exhibited larger MTBE adsorption uptakes than the tested GAC. Surface diffusion coefficients describing intraparticle MTBE mass transfer rates were largest for the GAC and smallest for the carbonaceous resin. Pilot tests were conducted to verify MTBE breakthrough curve predictions obtained with the homogeneous surface diffusion model and to evaluate the effect of NOM preloading on packed bed adsorber performance. Results showed that GAC was the most cost-competitive adsorbent when considering adsorbent usage rate only; however, the useful life of an adsorber containing silicalite zeolite was predicted to be approximately 5-6 times longer than that of an equally sized adsorber containing GAC. Pilot column results also showed that NOM preloading did not impair the MTBE removal efficiency of the silicalite zeolite. Thus, it may be possible to regenerate spent silicalite with less energy-intensive methods than those required to regenerate GAC.

  14. Edible films and coatings from whey proteins: a review on formulation, and on mechanical and bioactive properties.

    PubMed

    Ramos, Oscar L; Fernandes, João C; Silva, Sara I; Pintado, Manuela E; Malcata, F Xavier

    2012-01-01

    The latest decade has witnessed joint efforts by the packaging and the food industries to reduce the amount of residues and wastes associated with food consumption. The recent increase in environmental awareness has also contributed toward development of edible packaging materials. Viable edible films and coatings have been successfully produced from whey proteins; their ability to serve other functions, viz. carrier of antimicrobials, antioxidants, or other nutraceuticals, without significantly compromising the desirable primary barrier and mechanical properties as packaging films, will add value for eventual commercial applications. These points are tackled in this review, in a critical manner. The supply of whey protein-based films and coatings, formulated to specifically address end-user needs, is also considered.

  15. Adaptive hydrophobic and hydrophilic interactions of mussel foot proteins with organic thin films.

    PubMed

    Yu, Jing; Kan, Yajing; Rapp, Michael; Danner, Eric; Wei, Wei; Das, Saurabh; Miller, Dusty R; Chen, Yunfei; Waite, J Herbert; Israelachvili, Jacob N

    2013-09-24

    The adhesion of mussel foot proteins (Mfps) to a variety of specially engineered mineral and metal oxide surfaces has previously been investigated extensively, but the relevance of these studies to adhesion in biological environments remains unknown. Most solid surfaces exposed to seawater or physiological fluids become fouled by organic conditioning films and biofilms within minutes. Understanding the binding mechanisms of Mfps to organic films with known chemical and physical properties therefore is of considerable theoretical and practical interest. Using self-assembled monolayers (SAMs) on atomically smooth gold substrates and the surface forces apparatus, we explored the force-distance profiles and adhesion energies of three different Mfps, Mfp-1, Mfp-3, and Mfp-5, on (i) hydrophobic methyl (CH3)- and (ii) hydrophilic alcohol (OH)-terminated SAM surfaces between pH 3 and pH 7.5. At acidic pH, all three Mfps adhered strongly to the CH3-terminated SAM surfaces via hydrophobic interactions (range of adhesive interaction energy = -4 to -9 mJ/m(2)) but only weakly to the OH-terminated SAM surfaces through H- bonding (adhesive interaction energy ≤ -0.5 mJ/m(2)). 3, 4-Dihydroxyphenylalanine (Dopa) residues in Mfps mediate binding to both SAM surface types but do so through different interactions: typical bidentate H-bonding by Dopa is frustrated by the longer spacing of OH-SAMs; in contrast, on CH3-SAMs, Dopa in synergy with other nonpolar residues partitions to the hydrophobic surface. Asymmetry in the distribution of hydrophobic residues in intrinsically unstructured proteins, the distortion of bond geometry between H-bonding surfaces, and the manipulation of physisorbed binding lifetimes represent important concepts for the design of adhesive and nonfouling surfaces.

  16. Adaptive hydrophobic and hydrophilic interactions of mussel foot proteins with organic thin films

    PubMed Central

    Yu, Jing; Kan, Yajing; Rapp, Michael; Danner, Eric; Wei, Wei; Das, Saurabh; Miller, Dusty R.; Chen, Yunfei; Waite, J. Herbert; Israelachvili, Jacob N.

    2013-01-01

    The adhesion of mussel foot proteins (Mfps) to a variety of specially engineered mineral and metal oxide surfaces has previously been investigated extensively, but the relevance of these studies to adhesion in biological environments remains unknown. Most solid surfaces exposed to seawater or physiological fluids become fouled by organic conditioning films and biofilms within minutes. Understanding the binding mechanisms of Mfps to organic films with known chemical and physical properties therefore is of considerable theoretical and practical interest. Using self-assembled monolayers (SAMs) on atomically smooth gold substrates and the surface forces apparatus, we explored the force–distance profiles and adhesion energies of three different Mfps, Mfp-1, Mfp-3, and Mfp-5, on (i) hydrophobic methyl (CH3)- and (ii) hydrophilic alcohol (OH)-terminated SAM surfaces between pH 3 and pH 7.5. At acidic pH, all three Mfps adhered strongly to the CH3-terminated SAM surfaces via hydrophobic interactions (range of adhesive interaction energy = −4 to −9 mJ/m2) but only weakly to the OH-terminated SAM surfaces through H- bonding (adhesive interaction energy ≤ −0.5 mJ/m2). 3, 4-Dihydroxyphenylalanine (Dopa) residues in Mfps mediate binding to both SAM surface types but do so through different interactions: typical bidentate H-bonding by Dopa is frustrated by the longer spacing of OH-SAMs; in contrast, on CH3-SAMs, Dopa in synergy with other nonpolar residues partitions to the hydrophobic surface. Asymmetry in the distribution of hydrophobic residues in intrinsically unstructured proteins, the distortion of bond geometry between H-bonding surfaces, and the manipulation of physisorbed binding lifetimes represent important concepts for the design of adhesive and nonfouling surfaces. PMID:24014592

  17. Preparation and characterization of bio-nanocomposite films based on soy protein isolate and montmorillonite using melt extrusion

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The non-biodegradable and non-renewable nature of plastic packaging has led to a renewed interest in packaging materials based on bio-nanocomposites (biopolymer matrix reinforced with nanoparticles such as layered silicates). Bio-nanocomposite films based on soy protein isolate (SPI) and montmorillo...

  18. Gamma-irradiation influence on the structure and properties of calcium caseinate-whey protein isolate based films. Part 1. Radiation effect on the structure of proteins gels and films.

    PubMed

    Cieśla, Krystyna; Salmieri, Stephane; Lacroix, Monique

    2006-08-23

    Brookfield viscosimetry, Fourier transform infrared spectroscopy, transmission electron microscopy (TEM), and measurements of the texture strength of gels formed with CaCl2 and the mechanical and barrier properties of the film were applied in studies of gel formation and structural and mechanical properties of gels and films prepared using calcium caseinate (CC)-whey protein isolate (WPI)-glycerol (1:1:1), control, and irradiated with 60Co gamma rays using a 32 kGy dose. The irradiated gels have appeared to be more "fine-stranded" as compared to the more "particulate" control gels and lead to the formation of more rigid films with improved mechanical strength and barrier properties. This results from cross-linking and the modification of protein conformations were induced by irradiation, in particular the increase in the beta-sheet and beta-strand contents. Structural modifications taking place in CC-WPI composition are related to modifications taking place separately in CC and WPI. Improvement of the properties of the films after irradiation corresponds to the increased density of the cross-linked material because no change in the porosity of the films was observed by TEM.

  19. A comparison of adsorbed and grafted fibronectin coatings under static and dynamic conditions.

    PubMed

    Montaño-Machado, Vanessa; Hugoni, Ludivine; Díaz-Rodríguez, Sergio; Tolouei, Ranna; Chevallier, Pascale; Pauthe, Emmanuel; Mantovani, Diego

    2016-09-21

    Coatings for medical devices are expected to improve their surface biocompatibility mainly by being bioactive, i.e. stimulating healing-oriented interactions with living cells, tissues and organs. In particular, for stent applications, coatings are often designed to enhance the endothelialization process. The coating strategy will be primarily responsible for the interfacial properties between the substrate and the coating, which must show high stability. Therefore, the present work aims at comparing the stability of adsorbed and grafted fibronectin, a protein well-known to promote endothelialization. Fibronectin coatings were deposited on fluorocarbon films generated by a plasma-based process on stainless steel substrates. Then, deformation tests were performed in order to simulate the stenting procedure and stability tests were completed under static and under-flow conditions. Coatings were characterized by XPS, AFM, water contact angle, immunostaining and ToF-SIMS analyses. The results show higher stability for the grafted coatings; indeed, the integrity of the protein simply adsorbed was strongly compromised especially after under-flow tests. Both coatings exhibited similar behavior after deformation and static tests. These results clearly show the impact of the coating strategy on the overall stability of the coatings as well as the importance of under-flow investigations.

  20. Design and characterization of controlled-release edible packaging films prepared with synergistic whey-protein polysaccharide complexes.

    PubMed

    Liu, Fei; Jiang, Yanfeng; Du, Bingjian; Chai, Zhi; Jiao, Tong; Zhang, Chunyue; Ren, Fazheng; Leng, Xiaojing

    2013-06-19

    This paper describes an investigation into the properties of a doubly emulsified film incorporated with protein-polysaccharide microcapsules, which serves as a multifunctional food packaging film prepared using common edible materials in place of petroleum--based plastics. The relationships between the microstructural properties and controlled release features of a series of water-in-oil-in-water (W/O/W) microcapsulated edible films prepared in thermodynamically incompatible conditions were analyzed. The hydrophilic riboflavin (V(B2)) nano-droplets (13-50 nm) dispersed in α-tocopherol (V(E)) oil phase were embedded in whey protein-polysaccharide (WPs) microcapsules with a shell thickness of 20-56 nm. These microcapsules were then integrated in 103 μm thick WPs films. Different polysaccharides, including gum arabic (GA), low-methoxyl pectin (LMP), and κ-carrageenan (KCG), exhibited different in vitro synergistic effects on the ability of both films to effect enteric controlled release of both vitamins. GA, which showed a strong emulsifying ability, also showed better control of V(E) than other polysaccharides, and the highly charged KCG showed better control of V(B2) than GA did.

  1. Antimicrobial activity of whey protein films supplemented with Lactobacillus sakei cell-free supernatant on fresh beef.

    PubMed

    Beristain-Bauza, Silvia Del Carmen; Mani-López, Emma; Palou, Enrique; López-Malo, Aurelio

    2017-04-01

    The aim of this work was to evaluate the antimicrobial activity of whey protein isolate (WPI) films supplemented with Lactobacillus sakei NRRL B-1917 cell-free supernatant on beef inoculated with Escherichia coli ATCC 25922 or Listeria monocytogenes Scott A; additionally, sensory evaluation was performed on wrapped beef cubes. Supernatant concentrates were obtained from Lb. sakei cultures in MRS broth after centrifugation, filtering, and freeze-drying. Films were prepared with WPI (3% w/w), alginate (0.625% w/w), rehydrated supernatant (18 mg/ml), and glycerol. Films were used to wrap beef cubes inoculated with ≈10(3) CFU/g E. coli or L. monocytogenes. Sensory evaluation was carried out on grilled beef wrapped or not with the studied antimicrobial films. During refrigerated storage, antimicrobial films reduced 1.4 log10 CFU/g of L. monocytogenes after 120 h, while E. coli decreased 2.3 log10 CFU/g after 36 h. Grilled beef wrapped with antimicrobial film was well accepted by panelists, besides scores evidenced no significant differences (p > 0.05) between wrapped and unwrapped beef.

  2. Combinations of fluorescently labeled pulmonary surfactant proteins SP-B and SP-C in phospholipid films.

    PubMed Central

    Nag, K; Taneva, S G; Perez-Gil, J; Cruz, A; Keough, K M

    1997-01-01

    Hydrophobic pulmonary surfactant (PS) proteins B (SP-B) and C (SP-C) modulate the surface properties of PS lipids. Epifluorescence microscopy was performed on solvent-spread monolayers of fluorescently labeled porcine SP-B (R-SP-B, labeled with Texas Red) and SP-C (F-SP-C, labeled with fluorescein) in dipalmitoylphosphatidylcholine (DPPC) (at protein concentrations of 10 and 20 wt%, and 10 wt% of both) under conditions of cyclic compression and expansion. Matrix-assisted laser desorption/ionization (MALDI) spectroscopy of R-SP-B and F-SP-C indicated that the proteins were intact and labeled with the appropriate fluorescent probe. The monolayers were compressed and expanded for four cycles at an initial rate of 0.64 A2 x mol(-1) x s(-1) (333 mm2 x s x [-1]) up to a surface pressure pi approximately 65 mN/m, and pi-area per residue (pi-A) isotherms at 22 +/- 1 degrees C were obtained. The monolayers were microscopically observed for the fluorescence emission of the individual proteins present in the film lipid matrix, and their visual features were video recorded for image analysis. The pi-A isotherms of the DPPC/protein monolayers showed characteristic "squeeze out" effects at pi approximately 43 mN/m for R-SP-B and 55 mN/m for F-SP-C, as had previously been observed for monolayers of the native proteins in DPPC. Both proteins associated with the expanded (fluid) phase of DPPC monolayers remained in or associated with the monolayers at high pi (approximately 65 mN/m) and redispersed in the monolayer upon its reexpansion. At comparable pi and area/molecule of the lipid, the proteins reduced the amounts of condensed (gel-like) phase of DPPC monolayers, with F-SP-C having a greater effect on a weight basis than did R-SP-B. In any one of the lipid/protein monolayers the amounts of the DPPC in condensed phase were the same at equivalent pi during compression and expansion and from cycle to cycle. This indicated that only minor loss of components from these systems

  3. Rotary adsorbers for continuous bulk separations

    DOEpatents

    Baker, Frederick S [Oak Ridge, TN

    2011-11-08

    A rotary adsorber for continuous bulk separations is disclosed. The rotary adsorber includes an adsorption zone in fluid communication with an influent adsorption fluid stream, and a desorption zone in fluid communication with a desorption fluid stream. The fluid streams may be gas streams or liquid streams. The rotary adsorber includes one or more adsorption blocks including adsorbent structure(s). The adsorbent structure adsorbs the target species that is to be separated from the influent fluid stream. The apparatus includes a rotary wheel for moving each adsorption block through the adsorption zone and the desorption zone. A desorption circuit passes an electrical current through the adsorbent structure in the desorption zone to desorb the species from the adsorbent structure. The adsorbent structure may include porous activated carbon fibers aligned with their longitudinal axis essentially parallel to the flow direction of the desorption fluid stream. The adsorbent structure may be an inherently electrically-conductive honeycomb structure.

  4. Effect of surface charge distribution on the adsorption orientation of proteins to lipid monolayers.

    PubMed

    Tiemeyer, Sebastian; Paulus, Michael; Tolan, Metin

    2010-09-07

    The adsorption orientation of the proteins lysozyme and ribonuclease A (RNase A) to a neutral 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and a negatively charged stearic acid lipid film was investigated by means of X-ray reflectivity. Both proteins adsorbed to the negatively charged lipid monolayer, whereas at the neutral monolayer, no adsorption was observed. For acquiring comprehensive information on the proteins' adsorption, X-ray reflectivity data were combined with electron densities obtained from crystallographic data. With this method, it is possible to determine the orientation of adsorbed proteins in solution underneath lipid monolayers. While RNase A specifically coupled with its positively charged active site to the negatively charged lipid monolayer, lysozyme prefers an orientation with its long axis parallel to the Langmuir film. In comparison to the electrostatic maps of the proteins, our results can be explained by the discriminative surface charge distribution of lysozyme and RNase A.

  5. Long-Wavelength Infrared Sensing by Cytochrome C Protein Thin Film Deposited by the Spin Coating Method

    PubMed Central

    Lai, Bo-Yu; Chu, Chung-Hao; Su, Guo-Dung John

    2013-01-01

    High infrared absorption, large temperature coefficient of resistance (TCR) and small 1/f noise are preferred characteristics for sensing materials used in bolometers. In this paper, we discuss a cytochrome c protein as a potential sensing material for long-wavelength bolometers. We simulated and experimentally proved high infrared absorption of cytochrome c in the wavelength between 8 μm and 14 μm. Cytochrome c thin films were deposited on a hydrophilic surface using the spin coating method. The resistance variation with temperature is measured and we show that the TCR of cytochrome c thin films is consistently higher than 20%. The measured values of 1/f noise were as low as 2.33 × 10−13 V2/Hz at 60 Hz. Finally, we test the reliability of cytochrome c by measuring the resistance changes over time under varying conditions. We found that cytochrome c thin films deteriorated significantly without appropriate packaging. PMID:24264331

  6. Long-wavelength infrared sensing by cytochrome C protein thin film deposited by the spin coating method.

    PubMed

    Lai, Bo-Yu; Chu, Chung-Hao; Su, Guo-Dung John

    2013-11-20

    High infrared absorption, large temperature coefficient of resistance (TCR) and small 1/f noise are preferred characteristics for sensing materials used in bolometers. In this paper, we discuss a cytochrome c protein as a potential sensing material for long-wavelength bolometers. We simulated and experimentally proved high infrared absorption of cytochrome c in the wavelength between 8 μm and 14 μm. Cytochrome c thin films were deposited on a hydrophilic surface using the spin coating method. The resistance variation with temperature is measured and we show that the TCR of cytochrome c thin films is consistently higher than 20%. The measured values of 1/f noise were as low as 2.33 × 10⁻¹³ V²/Hz at 60 Hz. Finally, we test the reliability of cytochrome c by measuring the resistance changes over time under varying conditions. We found that cytochrome c thin films deteriorated significantly without appropriate packaging.

  7. Nanosize electropositive fibrous adsorbent

    DOEpatents

    Tepper, Frederick; Kaledin, Leonid

    2005-01-04

    Aluminum hydroxide fibers approximately 2 nanometers in diameter and with surface areas ranging from 200 to 650 m.sup.2 /g have been fount to be highly electropositive. When dispersed in water they are able to attach to and retain electronegative particles. When combined into a composite filter with other fibers or particles they can filter bacteria and nano size particulates such as viruses and colloidal particles at high flux through the filter. Such filters can be used for purification and sterilization of water, biological, medical and pharmaceutical fluids, and as a collector/concentrator for detection and assay of mirobes and viruses. The alumina fibers are also capable of filtering sub-micron inorganic and metallic particles to produce ultra pure water. The fibers are suitable as a substrate for growth of cells. Macromolicules such as proteins may be separated from each other based on their electronegative charges.

  8. Physical and mechanical properties in biodegradable films of whey protein concentrate-pullulan by application of beeswax.

    PubMed

    Khanzadi, Mehrdad; Jafari, Seid Mahdi; Mirzaei, Habibollah; Chegini, Faramarz Khodaian; Maghsoudlou, Yayha; Dehnad, Danial

    2015-03-15

    Different ratios of whey protein concentrate (WPC):pullulan (PUL) (70:30, 50:50, 30:70%w/w) and various rates of beeswax (BW) (0, 10, 20, and 30%w/wglycerol) were applied to prepare biodegradable WPC-PUL films containing glycerol as a plasticizer, for the first time. Thickness, moisture content, water solubility, water vapour permeability, colour, and mechanical properties of prepared films were measured. Higher ratios of WPC:PUL led to more desirable physical and mechanical properties; in other words, lower rates of thickness, moisture content, water solubility and water vapour permeability, and higher elongations were achieved. Application of BW (especially in higher contents) could successfully improve colour indices, diminish water solubility (nearly 12%) and water vapour permeability (approximately 3×10(-11)gm(-1)s(-1)Pa(-1)), and increase tensile strength (by about 7MPa) of WPC-PUL blend films. Our edible films enjoyed great whiteness and ignorable yellowness indices, making it a suitable alternative for application in food products. Overall, WPC70-PUL30 containing 30% BW resulted in the best performance of physical and mechanical aspects as an optimum film.

  9. Combined atomic force microscopy and spectroscopic ellipsometry applied to the analysis of lipid-protein thin films.

    PubMed

    Finot, Eric; Markey, Laurent; Hane, Francis; Amrein, Mathias; Leonenko, Zoya

    2013-04-01

    Pulmonary surfactant is a complex mixture of phospholipids and proteins and forms a thin film at the lung alveolar interface separating air from liquid environment. The film reduces the work of breathing during repeatable compressions of the alveoli which form a characteristic multilayer upon compression. In this work, we investigated the structure of bovine lipid extract surfactant (BLES). We analysed the BLES films by atomic force microscopy (AFM) and spectroscopic ellipsometry (SE) in order to provide combined characterization of both morphology and thickness of surfactant films. We show how the spectroscopic ellipsometry can be used to supplement the data obtained by AFM. We demonstrate that indium tin oxide (ITO) substrate used for spectroscopic ellipsometry is preferable over glass substrate to enhance the optical contrast. An optical model was proposed to account for non-uniform film morphology. We obtained good correlations between the multilayer surface coverage, determined by both AFM and SE. SE measures the thickness of the first uniform monolayer as 2.6 nm that cannot be achieved by AFM imaging alone.

  10. Thin films and assemblies of photosensitive membrane proteins and colloidal nanocrystals for engineering of hybrid materials with advanced properties.

    PubMed

    Zaitsev, Sergei Yu; Solovyeva, Daria O; Nabiev, Igor

    2012-11-15

    The development and study of nano-bio hybrid materials engineered from membrane proteins (the key functional elements of various biomembranes) and nanoheterostructures (inorganic colloidal nanoparticles, transparent electrodes, and films) is a rapidly growing field at the interface of materials and life sciences. The mainspring of the development of bioinspired materials and devices is the fact that biological evolution has solved many problems similar to those that humans are attempting to solve in the field of light-harvesting and energy-transferring inorganic compounds. Along this way, bioelectronics and biophotonics have shown considerable promise. A number of proteins have been explored in terms of bioelectronic device applications, but bacteriorhodopsin (bR, a photosensitive membrane protein from purple membranes of the bacterium Halobacterium salinarum) and bacterial photosynthetic reaction centres have received the most attention. The energy harvesting in plants has a maximum efficiency of 5%, whereas bR, in the absence of a specific light-harvesting system, allows bacteria to utilize only 0.1-0.5% of the solar light. Recent nano-bioengineering approaches employing colloidal semiconductor and metal nanoparticles conjugated with biosystems permit the enhancement of the light-harvesting capacity of photosensitive proteins, thus providing a strong impetus to protein-based device optimisation. Fabrication of ultrathin and highly oriented films from biological membranes and photosensitive proteins is the key task for prospective bioelectronic and biophotonic applications. In this review, the main advances in techniques of preparation of such films are analyzed. Comparison of the techniques for obtaining thin films leads to the conclusion that the homogeneity and orientation of biomembrane fragments or proteins in these films depend on the method of their fabrication and increase in the following order: electrophoretic sedimentation < Langmuir-Blodgett and

  11. Chitosan-whey protein edible films produced in the absence or presence of transglutaminase: analysis of their mechanical and barrier properties.

    PubMed

    Di Pierro, Prospero; Chico, Belkis; Villalonga, Reynaldo; Mariniello, Loredana; Damiao, Angelo E; Masi, Paolo; Porta, Raffaele

    2006-03-01

    Chitosan-whey protein edible films with different protein concentrations were prepared in the absence or presence of microbial transglutaminase as cross-linking agent. The films prepared in the presence of the enzyme showed low solubility at a wide range of pH, a lower degree of swelling, and good biodegradability following protease treatments. The presence of transglutaminase induced also an enhancement in film mechanical resistance and a reduction in their deformability. Finally, the barrier efficiency toward oxygen and carbon dioxide was found to be markedly improved in the cross-linked films which showed also a lower permeability to water vapor. Some potential practical applications of transglutaminase-treated chitosan-whey protein films are suggested.

  12. Allantoin as a solid phase adsorbent for removing endotoxins.

    PubMed

    Vagenende, Vincent; Ching, Tim-Jang; Chua, Rui-Jing; Gagnon, Pete

    2013-10-04

    In this study we present a simple and robust method for removing endotoxins from protein solutions by using crystals of the small-molecule compound 2,5-dioxo-4-imidazolidinyl urea (allantoin) as a solid phase adsorbent. Allantoin crystalline powder is added to a protein solution at supersaturated concentrations, endotoxins bind and undissolved allantoin crystals with bound endotoxins are removed by filtration or centrifugation. This method removes an average of 99.98% endotoxin for 20 test proteins. The average protein recovery is ∼80%. Endotoxin binding is largely independent of pH, conductivity, reducing agent and various organic solvents. This is consistent with a hydrogen-bond based binding mechanism. Allantoin does not affect protein activity and stability, and the use of allantoin as a solid phase adsorbent provides better endotoxin removal than anion exchange, polymixin affinity and biological affinity methods for endotoxin clearance.

  13. Innovative bionanocomposite films of edible proteins containing liposome-encapsulated nisin and halloysite nanoclay.

    PubMed

    Boelter, Juliana Ferreira; Brandelli, Adriano

    2016-09-01

    Films and coatings based on natural polymers have gained increased interest for food packaging applications. In this work, halloysite and phosphatidylcholine liposomes encapsulating nisin were used to develop nanocomposite films of gelatin and casein. Liposomes prepared with either soybean lecithin or Phospholipon(®) showed particle size ranging from 124 to 178nm and high entrapment efficiency (94-100%). Considering their stability, Phospholipon(®) liposomes with 1.0mg/ml nisin were selected for incorporation into nanocomposite films containing 0.5g/l halloysite. The films presented antimicrobial activity against Listeria monocytogenes, Clostridium perfringens and Bacillus cereus. Scanning electron microscopy revealed that the films had a smooth surface, but showed increased roughness with addition of liposomes and halloysite. Casein films were thinner and slightly yellowish, less rigid and very elastic as compared with gelatin films. Thermogravimetric analysis showed a decrease of the degradation temperature for casein films added with liposomes. The glass transition temperature decreased with addition of liposomes and halloysite. Gelatin and casein films containing nisin-loaded liposomes and halloysite represent an interesting alternative for development of active food packaging.

  14. Protein characterisation of Brosimum gaudichaudii Trécul latex and study of nanostructured latex film formation.

    PubMed

    Barbosa, Eduardo F; Monge-Fuentes, Victoria; Oliveira, Natiela B; Tavares, Rebecca; Xavier, Mary-Ann E; Bemquerer, Marcelo Porto; Silva, Luciano P

    2014-12-01

    Brosimum gaudichaudii Tréc. (Moraceae) is a common Brazilian Cerrado plant known by its pharmaceutical industry relevance. The authors investigated the latex protein components and potential biotechnological applications. Some protein fragments had their sequences elucidated, presenting similarities to jacalin and Kunitz-type trypsin inhibitors. Amino acid residue modifications were found, such as glutamine N-terminal residue cyclisation into pyroglutamic acid residue, and mass differences corresponding to hexoses and N-acetylhexosamine presence. The latex was used to produce a nanoscale structured film, which presented an increased attraction and reduced adhesion behaviours. The film presented high homogeneity, as observed by low nanoroughness values, probably because of its intrinsic components, such as the jacalin-like protein that has known agglutination properties. The immobilised Kunitz-type trypsin inhibitor presence in the latex film allow us to point out to applications related to this inhibition, as in active food packaging, since these peptidase inhibitors are able to inhibit pests and microorganism proliferation.

  15. Solution-based Syntheses of Iron Pyrite Thin Films for Photovoltaic and Protein Foot-printing Applications

    NASA Astrophysics Data System (ADS)

    El Makkaoui, Mohammed

    Iron pyrite (cubic FeS2) is a non-toxic, earth abundant semiconductor possessing a set of excellent optical/electronic properties for serving as an absorber layer in PV devices. Additionally, pyrite is a very efficient hydroxyl radical generator via Fenton chemistry and has shown promise in oxidative protein and DNA foot-printing application. The main focus of this thesis is on fabricating phase and elementally pure iron pyrite thin films using a solution-based approach that employs hydrazine as a solvent. A precursor ink is formed at room temperature by mixing elemental iron and sulfur in anhydrous hydrazine and then deposited on Mo-coated glass substrates, via spin coating, to yield amorphous iron sulfide films that are then annealed in H2S (340°C) and sulfur gas (≤ 500 °C) to form uniform, polycrystalline and phase pure pyrite films with densely packed grains. This approach is likely to yield the most elementally pure pyrite thin films made to date, through a very simple and scalable process. The ink has shown to be very sensitive to environmental conditions and has a very short shelf life (˜1 day). Additionally, the film microstructure is greatly influenced by the S:Fe concentration ratio that when tuned to 3:1, yielded uniform, robust and optically flat iron sulfide thin films with an optimal thickness (˜320 nm) for PV application. The results however were not reproducible, mainly due to failure in applying multiple layers without compromising film morphology. Thinner (< 100 nm) iron sulfide films, on the other hand, are reproducibly produced, but are too thin to be employed in PV devices. Direct annealing in sulfur gas at 475°C for 4 hours, bypassing the > 12 hour H2S annealing step, yielded phase pure pyrite films, with good morphology, at lower processing time and annealing temperatures (< 500°C). The latter part of this thesis regards the use of pyrite nano-crystals in conjunction with high surface area polymer laminates for protein foot

  16. Mysterious Lattice Rotations in Adsorbed Monolayers

    NASA Astrophysics Data System (ADS)

    Diehl, Renee D.

    1997-03-01

    Lattice rotations due to a mismatch in structure have been observed in film growth for many years, probably beginning in the 1930's with the Nishiyama-Wasserman and Kurdjumov-Sachs orientations observed when fcc(111) films grow on bcc(110) surfaces, or vice versa. Early analysis of this problem was carried out with the aid of Moiré patterns and the observation that the preferred lattice orientations are those which maximize the Moiré fringe spacing. Later energy calculations indicated that the structures which were predicted by the the Moiré technique actually do correspond to energy minima. Epitaxial rotation in adsorbed monolayers is a conceptually simpler problem since in principle it involves only two planes of atoms, and it was first observed in 1977 for Ar on a graphite surface(C. G. Shaw, M. D. Chinn, S. C. Fain, Jr. Phys. Rev. Lett. 41 (1978) 955.). This observation came only a few months after a new theory, based on the expected elastic behavior of an overlayer, was developed by A. D. Novaco and J. P. McTague(A. D. Novaco and J. P. McTague, Phys. Rev. Lett. 38 (1977) 1286.), and the agreement with the experimental results was remarkable. It was later shown that a few symmetry principles similar to those used for the film growth studies sometimes can also predict the observed structures. However, the situation for incommensurate layers physisorbed on metal surfaces currently looks bleak. None of the existing theories or models appears to describe the experimental results. New data for physisorbed gases on metal surfaces will be presented, along with some half-baked (and probably wrong) ideas for what might be happening. This work was supported by NSF.

  17. Development of SH-SAW sensors for measurement of the properties of protein solutions

    NASA Astrophysics Data System (ADS)

    Roh, Yongrae; Kwon, Yongjun; Kim, Kyungho; Koh, Kwangnak; Kim, Jaeho

    2003-07-01

    We developed new surface acoustic wave (SAW) sensors to measure the properties of protein solutions applying a particular organic thin film on the delay line of transverse type SAW devices. Each delay line is configured as an oscillator. The delay line for a sensing channel is coated with a gold film on which an antibody layer is immobilized by protein A. The sensing delay line selectively adsorbs antigens when exposed to a protein solution, which results in phase delay changes due to the mass loading effects induced by the adsorbed antigens. The other delay line is uncoated for use as a stable reference. The relative change in the frequency of the two oscillators is monitored to measure the concentration of the antigens. Sensor properties investigated include selectivity, sensitivity, response time and stability in response to the antigen concentration as well as the viscosity and electrical conductivity of the protein solution.

  18. Fabricating electrospun cellulose nanofibre adsorbents for ion-exchange chromatography.

    PubMed

    Dods, Stewart R; Hardick, Oliver; Stevens, Bob; Bracewell, Daniel G

    2015-01-09

    Protein separation is an integral step in biopharmaceutical manufacture with diffusion-limited packed bed chromatography remaining the default choice for industry. Rapid bind-elute separation using convective mass transfer media offers advantages in productivity by operating at high flowrates. Electrospun nanofibre adsorbents are a non-woven fibre matrix of high surface area and porosity previously investigated as a bioseparation medium. The effects of compression and bed layers, and subsequent heat treatment after electrospinning cellulose acetate nanofibres were investigated using diethylaminoethyl (DEAE) or carboxylate (COO) functionalisations. Transbed pressures were measured and compared by compression load, COO adsorbents were 30%, 70% and 90% higher than DEAE for compressions 1, 5 and 10MPa, respectively, which was attributed to the swelling effect of hydrophilic COO groups. Dynamic binding capacities (DBCs) at 10% breakthrough were measured between 2000 and 12,000CV/h (2s and 0.3s residence times) under normal binding conditions, and DBCs increased with reactant concentration from 4 to 12mgBSA/mL for DEAE and from 10 to 21mglysozyme/mL for COO adsorbents. Comparing capacities of compression loads applied after electrospinning showed that the lowest load tested, 1MPa, yielded the highest DBCs for DEAE and COO adsorbents at 20mgBSA/mL and 27mglysozyme/mL, respectively. At 1MPa, DBCs were the highest for the lowest flowrate tested but stabilised for flowrates above 2000CV/h. For compression loads of 5MPa and 10MPa, adsorbents recorded lower DBCs than 1MPa as a result of nanofibre packing and reduced surface area. Increasing the number of bed layers from 4 to 12 showed decreasing DBCs for both adsorbents. Tensile strengths were recorded to indicate the mechanical robustness of the adsorbent and be related to packing the nanofibre adsorbents in large scale configurations such as pleated cartridges. Compared with an uncompressed adsorbent, compressions of 1, 5

  19. Fabricating electrospun cellulose nanofibre adsorbents for ion-exchange chromatography

    PubMed Central

    Dods, Stewart R.; Hardick, Oliver; Stevens, Bob; Bracewell, Daniel G.

    2015-01-01

    Protein separation is an integral step in biopharmaceutical manufacture with diffusion-limited packed bed chromatography remaining the default choice for industry. Rapid bind-elute separation using convective mass transfer media offers advantages in productivity by operating at high flowrates. Electrospun nanofibre adsorbents are a non-woven fibre matrix of high surface area and porosity previously investigated as a bioseparation medium. The effects of compression and bed layers, and subsequent heat treatment after electrospinning cellulose acetate nanofibres were investigated using diethylaminoethyl (DEAE) or carboxylate (COO) functionalisations. Transbed pressures were measured and compared by compression load, COO adsorbents were 30%, 70% and 90% higher than DEAE for compressions 1, 5 and 10 MPa, respectively, which was attributed to the swelling effect of hydrophilic COO groups. Dynamic binding capacities (DBCs) at 10% breakthrough were measured between 2000 and 12,000 CV/h (2 s and 0.3 s residence times) under normal binding conditions, and DBCs increased with reactant concentration from 4 to 12 mg BSA/mL for DEAE and from 10 to 21 mg lysozyme/mL for COO adsorbents. Comparing capacities of compression loads applied after electrospinning showed that the lowest load tested, 1 MPa, yielded the highest DBCs for DEAE and COO adsorbents at 20 mg BSA/mL and 27 mg lysozyme/mL, respectively. At 1 MPa, DBCs were the highest for the lowest flowrate tested but stabilised for flowrates above 2000 CV/h. For compression loads of 5 MPa and 10 MPa, adsorbents recorded lower DBCs than 1 MPa as a result of nanofibre packing and reduced surface area. Increasing the number of bed layers from 4 to 12 showed decreasing DBCs for both adsorbents. Tensile strengths were recorded to indicate the mechanical robustness of the adsorbent and be related to packing the nanofibre adsorbents in large scale configurations such as pleated cartridges. Compared with an

  20. The Investigation and Characterization of the Group 3 [Nickel-Iron]-Hydrogenases Using Protein Film Electrochemistry

    NASA Astrophysics Data System (ADS)

    McIntosh, Chelsea Lee

    Hydrogenases, the enzymes that reversibly convert protons and electrons to hydrogen, are used in all three domains of life. [NiFe]-hydrogenases are considered best suited for biotechnological applications because of their reversible inactivation with oxygen. Phylogenetically, there are four groups of [NiFe]-hydrogenases. The best characterized group, "uptake" hydrogenases, are membrane-bound and catalyze hydrogen oxidation in vivo. In contrast, the group 3 [NiFe]-hydrogenases are heteromultimeric, bifunctional enzymes that fulfill various cellular roles. In this dissertation, protein film electrochemistry (PFE) is used to characterize the catalytic properties of two group 3 [NiFe]-hydrogenases: HoxEFUYH from Synechocystsis sp. PCC 6803 and SHI from Pyrococcus furiosus. First, HoxEFUYH is shown to be biased towards hydrogen production. Upon exposure to oxygen, HoxEFUYH inactivates to two states, both of which can be reactivated on the timescale of seconds. Second, we show that PfSHI is the first example of an oxygen tolerant [NiFe]-hydrogenase that produces two inactive states upon exposure to oxygen. Both inactive states are analogous to those characterized for HoxEFUYH, but oxygen exposed PfSHI produces a greater fraction that reactivates at high potentials, enabling hydrogen oxidation in the presence of oxygen. Third, it is shown that removing the NAD(P)-reducing subunits from PfSHI leads to a decrease in bias towards hydrogen oxidation and renders the enzyme oxygen sensitive. Both traits are likely due to impaired intramolecular electron transfer. Mechanistic hypotheseses for these functional differences are considered.

  1. Deoxynivalenol impairs hepatic and intestinal gene expression of selected oxidative stress, tight junction and inflammation proteins in broiler chickens, but addition of an adsorbing agent shifts the effects to the distal parts of the small intestine.

    PubMed

    Osselaere, Ann; Santos, Regiane; Hautekiet, Veerle; De Backer, Patrick; Chiers, Koen; Ducatelle, Richard; Croubels, Siska

    2013-01-01

    Broiler chickens are rather resistant to deoxynivalenol and thus, clinical signs are rarely seen. However, effects of subclinical concentrations of deoxynivalenol on both the intestine and the liver are less frequently studied at the molecular level. During our study, we investigated the effects of three weeks of feeding deoxynivalenol on the gut wall morphology, intestinal barrier function and inflammation in broiler chickens. In addition, oxidative stress was evaluated in both the liver and intestine. Besides, the effect of a clay-based mycotoxin adsorbing agent on these different aspects was also studied. Our results show that feeding deoxynivalenol affects the gut wall morphology both in duodenum and jejenum of broiler chickens. A qRT-PCR analysis revealed that deoxynivalenol acts in a very specific way on the intestinal barrier, since only an up-regulation in mRNA expression of claudin 5 in jejunum was observed, while no effects were seen on claudin 1, zona occludens 1 and 2. Addition of an adsorbing agent resulted in an up-regulation of all the investigated genes coding for the intestinal barrier in the ileum. Up-regulation of Toll-like receptor 4 and two markers of oxidative stress (heme-oxigenase or HMOX and xanthine oxidoreductase or XOR) were mainly seen in the jejunum and to a lesser extent in the ileum in response to deoxynivalenol, while in combination with an adsorbing agent main effect was seen in the ileum. These results suggest that an adsorbing agent may lead to higher concentrations of deoxynivalenol in the more distal parts of the small intestine. In the liver, XOR was up-regulated due to DON exposure. HMOX and HIF-1α (hypoxia-inducible factor 1α) were down-regulated due to feeding DON but also due to feeding the adsorbing agent alone or in combination with DON.

  2. Modification of polyetherurethane for biomedical application by radiation induced grafting. II. Water sorption, surface properties, and protein adsorption of grafted films

    SciTech Connect

    Jansen, B.; Ellinghorst, G.

    1984-07-01

    A series of polyetherurethane films grafted by means of gamma radiation with hydrophilic or reactive monomers (2-hydroxyethyl methacrylate, 2,3-epoxypropyl methacrylate, 2,3-dihydroxypropyl methacrylate, and acrylamide) and partially chemically modified were subjected to various physico-chemical investigation methods involving water sorption, contact angle, and protein adsorption measurements. From contact angle data the interfacial free energy gamma sw between grafted films and water was calculated. It was found that the water uptake of grafted films increases with grafting yield or, in the case of grafted and afterwards chemically modified films, with reaction yield; the diffusion coefficient of water in the modified films also increases with grafting yield. Contact angle studies revealed all grafted films to have surfaces more hydrophilic than the ungrafted trunk polymer. The degree of hydrophilicity--especially of HEMA-grafted films--strongly depends on grafting conditions. For some grafted samples with high surface hydrophilicity very low interfacial free energies approaching zero were measured. The study of the competitive adsorption of bovine serum albumin, gamma-globulin, and fibrinogen from a synthetic protein solution onto modified films showed that the adsorption of albumin increases markedly with increasing grafting yields, whereas the fibrinogen and gamma-globulin adsorption only slightly increases. A correlation between interfacial free energy and protein adsorption in the sense of the minimum interfacial free energy hypothesis was found only for samples with grafting yields below 5%. At higher grafting yields the increased surface area complicates the analysis.

  3. Interactions of organic contaminants with mineral-adsorbed surfactants.

    PubMed

    Zhu, Lizhong; Chen, Baoliang; Tao, Shu; Chiou, Cary T

    2003-09-01

    Sorption of organic contaminants (phenol, p-nitrophenol, and naphthalene) to natural solids (soils and bentonite) with and without myristylpyridinium bromide (MPB) cationic surfactant was studied to provide novel insightto interactions of contaminants with the mineral-adsorbed surfactant. Contaminant sorption coefficients with mineral-adsorbed surfactants, Kss, show a strong dependence on surfactant loading in the solid. At low surfactant levels, the Kss values increased with increasing sorbed surfactant mass, reached a maximum, and then decreased with increasing surfactant loading. The Kss values for contaminants were always higher than respective partition coefficients with surfactant micelles (Kmc) and natural organic matter (Koc). At examined MPB concentrations in water the three organic contaminants showed little solubility enhancement by MPB. At low sorbed-surfactant levels, the resulting mineral-adsorbed surfactant via the cation-exchange process appears to form a thin organic film, which effectively "adsorbs" the contaminants, resulting in very high Kss values. At high surfactant levels, the sorbed surfactant on minerals appears to form a bulklike medium that behaves essentially as a partition phase (rather than an adsorptive surface), with the resulting Kss being significantly decreased and less dependent on the MPB loading. The results provide a reference to the use of surfactants for remediation of contaminated soils/sediments or groundwater in engineered surfactant-enhanced washing.

  4. Interactions of organic contaminants with mineral-adsorbed surfactants

    USGS Publications Warehouse

    Zhu, L.; Chen, B.; Tao, S.; Chiou, C.T.

    2003-01-01

    Sorption of organic contaminants (phenol, p-nitrophenol, and naphthalene) to natural solids (soils and bentonite) with and without myristylpyridinium bromide (MPB) cationic surfactant was studied to provide novel insight to interactions of contaminants with the mineral-adsorbed surfactant. Contaminant sorption coefficients with mineral-adsorbed surfactants, Kss, show a strong dependence on surfactant loading in the solid. At low surfactant levels, the Kss values increased with increasing sorbed surfactant mass, reached a maximum, and then decreased with increasing surfactant loading. The Kss values for contaminants were always higher than respective partition coefficients with surfactant micelles (Kmc) and natural organic matter (Koc). At examined MPB concentrations in water the three organic contaminants showed little solubility enhancement by MPB. At low sorbed-surfactant levels, the resulting mineral-adsorbed surfactant via the cation-exchange process appears to form a thin organic film, which effectively "adsorbs" the contaminants, resulting in very high Kss values. At high surfactant levels, the sorbed surfactant on minerals appears to form a bulklike medium that behaves essentially as a partition phase (rather than an adsorptive surface), with the resulting Kss being significantly decreased and less dependent on the MPB loading. The results provide a reference to the use of surfactants for remediation of contaminated soils/sediments or groundwater in engineered surfactant-enhanced washing.

  5. Optimizing heterosurface adsorbent synthesis for liquid chromatography

    NASA Astrophysics Data System (ADS)

    Bogoslovskii, S. Yu.; Serdan, A. A.

    2016-03-01

    The structural and geometric parameters of a silica matrix (SM) for the synthesis of heterosurface adsorbents (HAs) are optimized. Modification is performed by shielding the external surfaces of alkyl-modified silica (AS) using human serum albumin and its subsequent crosslinking. The structural and geometric characteristics of the SM, AS, and HA are measured via low-temperature nitrogen adsorption. It is found that the structural characteristics of AS pores with diameters D < 6 nm do not change during HA synthesis, while the volume of pores with diameters of 6 nm < D < 9 nm shrinks slightly due to the adsorption of albumin in the pore orifices. It is established that the volume of pores with diameters D > 9 nm reduces significantly due to adsorption of albumin. It is concluded that silica gel with a maximum pore size distribution close to 5 nm and a minimal proportion of pores with D > 9 nm is optimal for HA synthesis; this allows us to achieve the greatest similarity between the chromatographic retention parameters for HA and AS. The suitability of the synthesized adsorbents for analyzing drugs in biological fluids through direct sample injection is confirmed by chromatography. It was found that the percentage of the protein fraction detected at the outlet of the chromatographic column is 98%.

  6. Effect of Sodium Sulfite, Sodium Dodecyl Sulfate, and Urea on the Molecular Interactions and Properties of Whey Protein Isolate-Based Films

    PubMed Central

    Schmid, Markus; Prinz, Tobias K.; Stäbler, Andreas; Sängerlaub, Sven

    2017-01-01

    Whey protein coatings and cast films are promising for use as food packaging materials. Ongoing research is endeavoring to reduce their permeability. The intention of this study was to evaluate the effect of the reactive additives sodium sulfite, sodium dodecyl sulfate (SDS), and urea on the oxygen barrier, water vapor barrier, and protein solubility of whey protein cast films. The concentration of the reactive additives was 1 to 20 wt.-%. Dried whey protein cast films were used as substrate materials. The water vapor transmission rate, the oxygen permeability, and the protein solubility were measured. Effective diffusion coefficients and effective sorption coefficients were calculated from the results of the water vapor sorption experiments. The presence of sodium sulfite resulted in an increased number of hydrophobic interactions and hydrogen bonds and a slightly decreased number of disulfide bonds. The oxygen permeability decreased from 68 to 46 cm3 (STP/standard temperature and pressure) 100 μm (m2 d bar)−1 for 1 wt.-% SDS in the whey protein cast film. The water vapor transmission rate decreased from 165 to 44 g 100 μm (m2 d)−1 measured at 50 to 0% r. h. for 20 wt.-% SDS in the whey protein cast film. The reduction in the water vapor transmission rate correlated with the lower effective diffusion coefficient. PMID:28149835

  7. Analysis and modeling of moisture sorption behavior for antimicrobial composite protein films.

    PubMed

    Lei, Qiao; Pan, Jiazhen; Bao, Jianqiang; Huang, Zhiying; Zhang, Yuting

    2014-01-01

    The WPI-NaCas-GLY antimicrobial film takes full advantage of the controlled release of active or antimicrobial agents as well as demonstrates a great potential for functioning as an alternative biodegradable polymer in practical applications. The moisture sorption kinetics of the film as an important carrier of active agents was investigated at various relative humidities (RH). The results indicated that the moisture sorption characterization and procedure of this film can be described well by the empirical Peleg model with higher confidence and concordance. The model could predict the film's moisture content at any time (Mt), the time to reach any given level of R (tR), the equilibrium moisture at any RH condition (Me), and isotherm trend based upon experimental data and modeled constants k(1), k(2), a, b, c, and d without giving consideration to their physical meaning. The water vapor transmission rate of the WPI-NaCas-GLY antimicrobial film increased exponentially with increasing RH due to its hydrophilicity, which was primarily caused by the presence of glycerol in a higher content. The results also suggested that aw predominately affects the film's Me values compared with the temperature factor by fixed nonlinear multiple regression analyses.

  8. Monte Carlo lattice models for adsorbed polymer conformation

    NASA Technical Reports Server (NTRS)

    Good, B. S.

    1985-01-01

    The adhesion between a polymer film and a metal surface is of great technological interest. However, the prediction of adhesion and wear properties of polymer coated metals is quite difficult because a fundamental understanding of the polymer surface interaction does not yet exist. A computer model for the conformation of a polymer molecule adsorbed on a surface is discussed. The chain conformation is assumed to be described by a partially directed random walk on a three dimensional simple cubic lattice. An attractive surface potential is incorporated into the model through the use of a random walk step probability distribution that is anisotropic in the direction normal to the attractive surface. The effects of variations in potential characteristics are qualitatively included by varying both the degree of anisotropy of the step distribution and the range of the anisotropy. Polymer conformation is characterized by the average end to end distance, average radius of gyration, and average number of chain segments adsorbed on the surface.

  9. Properties and microstructure of protein-based film from round scad (Decapterus maruadsi) muscle as affected by palm oil and chitosan incorporation.

    PubMed

    Prodpran, Thummanoon; Benjakul, Soottawat; Artharn, Anuchit

    2007-12-01

    The properties of protein-based film prepared from round scad (Decapterus maruadsi) muscle in the absence and the presence of palm oil and/or chitosan were investigated. Films added with 25% palm oil (as glycerol substitiution) had the slight decrease in water vapor permeability (WVP) and elongation at break (EAB) (p<0.05). WVP and tensile strength (TS) of films increased but EAB decreased when 10-40% chitosan (as protein substitution) was incorporated (p<0.05). Hydrophobic interactions and hydrogen bonds, together with disulfide and non-disulfide covalent bonds, played an important role in stabilizing the film matrix. The a* and b*-values increased with increasing chitosan levels (p<0.05). Films added with chitosan were less transparent and had the lowered transmission in the visible range. The incorporation of 25% palm oil and 40% chitosan yielded the films with the improved TS but decreased water vapor barrier property. Apart from film strengthening effect, chitosan inconjunction with Tween-20 most likely functioned as the emulsifier/stabilizer in film forming solution containing palm oil.

  10. Use of /γ-irradiation cross-linking to improve the water vapor permeability and the chemical stability of milk protein films

    NASA Astrophysics Data System (ADS)

    Ouattara, B.; Canh, L. T.; Vachon, C.; Mateescu, M. A.; Lacroix, M.

    2002-03-01

    γ-irradiation was used to produce free-standing cross-linked milk proteins. Film forming solutions were prepared according to a method previously developed in our laboratory using calcium caseinate (cas) with various proportions of whey protein isolate (wpi) or whey protein concentrate (wpc). The following caseinate-whey protein (cas:wp) ratio were prepared: 100:0, 75:25, 50:50, 25:75, and 0:100. The WVP of the films was determined gravimetrically at 23°C using a modified ASTM procedure. Molecular properties characterization was performed by size exclusion chromatography (SEC). Results showed significant ( p⩽0.05) reduction of the WVP of protein films for the following formulations: cas:wpi or cas:wpc (100:0); cas:wpi (25:75); cas:wpc (25:75); and cas:wpc (0:100). Mixture of cas and wpi produced a synergistic effect. The strongest combined effect was obtained for cas:wpi (25:75) formulation with permeability values of 2.07 and 1.38 g mm/m 2 d mm Hg for unirradiated and irradiated samples, respectively. γ-irradiation also induced a substantial increase of high molecular weight protein components in film forming solutions. The predominant fraction was ⩾10×10 6 Da for irradiated film forming solutions, compared to less than 0.2×10 6 Da for native unirradiated solutions.

  11. Effect of film thickness on the antifouling performance of poly(hydroxy-functional methacrylates) grafted surfaces.

    PubMed

    Zhao, Chao; Li, Lingyan; Wang, Qiuming; Yu, Qiuming; Zheng, Jie

    2011-04-19

    The development of nonfouling biomaterials to prevent nonspecific protein adsorption and cell/bacterial adhesion is critical for many biomedical applications, such as antithrombogenic implants and biosensors. In this work, we polymerize two types of hydroxy-functional methacrylates monomers of 2-hydroxyethyl methacrylate (HEMA) and hydroxypropyl methacrylate (HPMA) into polymer brushes on the gold substrate via surface-initiated atom transfer radical polymerization (SI-ATRP). We systematically examine the effect of the film thickness of polyHEMA and polyHPMA brushes on their antifouling performance in a wide range of biological media including single-protein solution, both diluted and undiluted human blood serum and plasma, and bacteria culture. Surface plasmon resonance (SPR) results show a strong correlation between antifouling property and film thickness. Too thin or too thick polymer brushes lead to large protein adsorption. Surfaces with the appropriate film thickness of ∼25-45 nm for polyHPMA and ∼20-45 nm for polyHEMA can achieve almost zero protein adsorption (<0.3 ng/cm(2)) from single-protein solution and diluted human blood plasma and serum. For undiluted human blood serum and plasma, polyHEMA brushes at a film thickness of ∼20-30 nm adsorb only ∼3.0 and ∼3.5 ng/cm(2) proteins, respectively, while polyHPMA brushes at a film thickness of ∼30 nm adsorb more proteins of ∼13.5 and ∼50.0 ng/cm(2), respectively. Moreover, both polyHEMA and polyHPMA brushes with optimal film thickness exhibit very low bacteria adhesion. The excellent antifouling ability and long-term stability of polyHEMA and polyHPMA brushes make them, especially for polyHEMA, effective and stable antifouling materials for usage in blood-contacting devices.

  12. Chitosan-whey protein isolate composite films for encapsulation and stabilization of fish oil containing ultra pure omega-3 fatty acids.

    PubMed

    Duan, Jingyun; Jiang, Yan; Zhao, Yanyun

    2011-01-01

    Chitosan (1.5%, w/v)-whey protein isolate (WPI, 5% w/v) composite films were developed for encapsulating and stabilizing fish oil (FO) containing 93.7% eicosapentaenoic acid (EPA). Chitosan-WPI film-forming solutions (FFS) were incorporated with 1.5% or 2% FO (w/v), 2% (w/v) glycerol, Tween 80 (3 times weight of FO), and 0.5% (w/v) oregano or rosemary essential oil (EO), and cast for films at room conditions. Dried films were stored at 2 °C for 30 d for evaluating encapsulation efficiency (EE), lipid stability, and film functionality. Total oil contents in films from FFS incorporating 1.5% or 2% FO were 28.1% to 32.5% and 33.4% to 37.3%, respectively, and free oil contents were 13.5% to 14.7% and 15.5% to 16.3%, respectively. EE, moisture content, and water activity of the films were 47.8% to 66%, 18.7% to 24.9%, and 0.42% to 0.50%, respectively, without significant difference among differently formulated films. Increasing FO concentration from 1.5% to 2% in FFS decreased tensile strength of the films from 0.57-0.73 to 0.34-0.44 MPa, but not the film elongation. Addition of oregano EO in FFS retarded lipid oxidation of the fish oil encapsulated in the films, in which a 43% to 53% reduction in thiobarbituric acid-reactive substances value and 39% to 51% reduction in peroxide value were achieved. Chitosan-WPI composite films with incorporation of oregano essential oil could be applied as a simple and economic means for encapsulating and stabilizing fish oil for fortifying omega-3 fatty acids in various applications.

  13. SPR-MS: from identifying adsorbed molecules to image tissues

    NASA Astrophysics Data System (ADS)

    Masson, Jean-François; Breault-Turcot, Julien; Forest, Simon; Chaurand, Pierre

    2015-03-01

    Surface plasmon resonance (SPR) sensors have become valuable analytical sensors for biomolecule detection. While SPR is heralded with high sensitivity, label-free and real-time detection, nonspecific adsorption and detection of ultralow concentrations remain issues. Nonspecific adsorption can be minimized using adequate surface chemistry. For example, we have employed peptide monolayers to reduce nonspecific adsorption of crude serum or cell lysate. It is important to uncover the nature of molecules nonspecifically adsorbing to surfaces in these biofluids, to further improve understanding of the nonspecific adsorption processes. Mass spectrometry (MS) provides a complementary tool to SPR to identify biomolecule adsorbed to surface. Trypsic digestion of the proteins adsorbed to surfaces led to identification of characteristic peptides from the proteins involved in nonspecific adsorption. Nonspecific adsorption in crude cell lysate results mainly from lipids, as confirmed with SPR and MS but proteins were observed on some surfaces. In another application of SPR and MS, imaging SPR can be used in combination to imaging MS to image tissue sections. Thin sections of mouse liver were inserted in the fluidic chamber of a SPRi instrument and proteins were transferred to the SPRi chip. The SPR chip was then imaged using MALDI imaging MS to identify the biomolecules that were transferred to the SPRi chip.

  14. Parallel pore and surface diffusion of levulinic acid in basic polymeric adsorbents.

    PubMed

    Liu, Baojian; Yang, Yiwen; Ren, Qilong

    2006-11-03

    The equilibrium and kinetics of levulinic acid (LA) adsorption on two basic polymeric adsorbents, 335 (highly porous gel) and D315 (macroreticular), were investigated. Experimental adsorption rates in batch stirred vessels under a variety of operating conditions were described successfully by the parallel pore and surface diffusion model taking into account external mass transfer and nonlinear Toth isotherm. The film-pore diffusion model was matched with the rate data and the resulting apparent pore diffusivities were strongly concentration-dependent and approached to a constant value for 335 adsorbent. Thus, the constant value was taken as the accurate pore diffusivity, while the pore diffusivity in D315 was estimated from the particle porosity. The surface diffusivities decreased with increasing initial bulk concentration for both adsorbents. The inverse concentration dependence was correlated reasonably well to the change of isosteric heat of adsorption as amount adsorbed.

  15. In vitro investigation of protein adsorption and platelet adhesion on inorganic biomaterial surfaces

    NASA Astrophysics Data System (ADS)

    Huang, Yan; Lü, Xiaoying; Jingwu, Ma; Huang, Nan

    2008-11-01

    The aim of this paper was to study the surface properties, protein adsorption and platelet adhesion behaviors of diamond-like carbon (DLC) and titanium (Ti) films. The surface energy and microstructures of these films were characterized by contact angle measurement and atomic force microscopy (AFM). A modified Coomassie brilliant blue (CBB) protein assay was used to study the amount of adsorbed proteins. Platelet adhesion was assessed by scanning electron microscopy (SEM). The AFM results show that the DLC film is smoother than Ti. Protein adsorption results from CBB protein assay show that the ratio of adsorbed albumin (Alb) to IgG ( RA/I) on DLC is larger than Ti, which coincide with the sequence of the ratio of interfacial tension between solid surface and Alb ( γS,Alb) to interfacial tension between surface and IgG ( γS,IgG) ( γS,Alb/ γS,IgG). The DLC film has a preferential adsorption for Alb. The results suggest that the ratio of γS,Alb/ γS,IgG may indicate an Alb/IgG affinity ratio of materials. More platelets adhere on Ti film than on DLC, which may correspond to the surface roughness of materials. The conclusion is the blood compatibility of DLC seems to be better than Ti.

  16. Physical properties of emulsion-based hydroxypropyl methylcellulose/whey protein isolate (HPMC/WPI) edible films.

    PubMed

    Rubilar, Javiera F; Zúñiga, Rommy N; Osorio, Fernando; Pedreschi, Franco

    2015-06-05

    The objective of this research was to study the effect of the film microstructure of oil-in-water emulsions stabilized by hydroxypropyl methyl cellulose/whey protein isolate (HPMC/WPI) with or without sodium dodecyl sulfate (SDS) over physical properties of HPMC/WPI emulsion-based films. The films were prepared with different HPMC/WPI-oil-SDS combinations (%w/w for 100g of dispersion): HPMC; WPI; HPMC/1WPI-0.5-SDS; HPMC/1WPI-1; HPMC/2WPI-0.5; HPMC/2WPI-1-SDS. Physical properties of films were evaluated. The results showed no statistical differences (p>0.05) between the thicknesses of EFs (0.156 ± 0.004 mm). The effect of oil content and incorporation of SDS showed the inverse trend for WI and ΔE, the increasing order of change, for WI and ΔE, among the formulation evaluated was: HPMC/1WPI-1>HPMC/2WPI-0.5>HPMC/2WPI-1.0-SDS≈HPMC/1WPI-0.5-SDS≈WPI>HPMC for WI and HPMC/1WPI-0.5-SDS>HPMC/2WPI-1.0-SDS>HPMC/2WPI-0.5>HPMC/1WPI-1 for ΔE, respectively. The addition of oil and SDS decreased the TS and EB, because oil addition into EF induces the development of structural discontinuities, producing an EF with less chain mobility, and consequently, with less flexibility and resistance to fracture.

  17. Protein adsorption on surfaces: dynamic contact-angle (DCA) and quartz-crystal microbalance (QCM) measurements.

    PubMed

    Stadler, H; Mondon, M; Ziegler, C

    2003-01-01

    Adsorption of the protein bovine serum albumin (BSA) on gold has been tested at various concentrations in aqueous solution by dynamic contact-angle analysis (DCA) and quartz-crystal microbalance (QCM) measurements. With the Wilhelmy plate technique advancing and receding contact angles and the corresponding hysteresis were measured and correlated with the hydrophilicity and the homogeneity of the surface. With electrical admittance measurements of a gold-coated piezoelectrical quartz crystal, layer mass and viscoelastic contributions to the resonator's frequency shift during adsorption could be separated. A correlation was found between the adsorbed mass and the homogeneity and hydrophilicity of the adsorbed film.

  18. Physicochemical properties of soy protein isolate/carboxymethyl cellulose blend films crosslinked by Maillard reactions: color, transparency and heat-sealing ability.

    PubMed

    Su, Jun-Feng; Yuan, Xiao-Yan; Huang, Zhen; Wang, Xin-Yu; Lu, Xu-Zhen; Zhang, Li-Dan; Wang, Sheng-Bao

    2012-01-01

    Soy protein isolate (SPI) films have many potential applications in the biomaterial field as surgical dressings for burns, films for reduction of wound inflammation, and facial masks. The appearance and the sealing ability are important physicochemical properties that greatly influence consumer acceptance of such protein-based films. The aim of the present work was to investigate the chemical structure and the physical properties associated with color, transparency and heat-sealing ability for SPI/carboxymethyl cellulose (CMC) blend films prepared by solution casting, with weight proportions 90/10, 80/20, 70/30 and 60/40. Fourier transform infra-red (FTIR) and solid-state (13)C nuclear magnetic resonance (NMR) spectra confirmed that Maillard reactions occurred between SPI and CMC. The Hunter color value (L, a, b) and transparency of films were affected by varying the proportions of SPI and CMC. With increasing degree of crosslinking of SPI and CMC, the yellow color of the films was diluted and transparency was improved. Peel strength and tensile strength measurements showed that the Maillard reactions had the main effect of enhancing the heat-sealing ability above the melting temperature. These results indicated that the structure and properties of SPI-based films could be modified and improved by blending with CMC.

  19. Guiding Principles of Hydrogenase Catalysis Instigated and Clarified by Protein Film Electrochemistry.

    PubMed

    Armstrong, Fraser A; Evans, Rhiannon M; Hexter, Suzannah V; Murphy, Bonnie J; Roessler, Maxie M; Wulff, Philip

    2016-05-17

    Protein film electrochemistry (PFE) is providing cutting-edge insight into the chemical principles underpinning biological hydrogen. Attached to an electrode, many enzymes exhibit "reversible" electrocatalytic behavior, meaning that a catalyzed redox reaction appears reversible or quasi-reversible when viewed by cyclic voltammetry. This efficiency is most relevant for enzymes that are inspiring advances in renewable energy, such as hydrogen-activating and CO2-reducing enzymes. Exploiting the rich repertoire of available instrumental methods, PFE experiments yield both a general snapshot and fine detail, all from tiny samples of enzyme. The dynamic electrochemical investigations blaze new trails and add exquisite detail to the information gained from structural and spectroscopic studies. This Account describes recent investigations of hydrogenases carried out in Oxford, including ideas initiated with PFE and followed through with complementary techniques, all contributing to an eventual complete picture of fast and efficient H2 activation without Pt. By immobilization of an enzyme on an electrode, catalytic electron flow and the chemistry controlling it can be addressed at the touch of a button. The buried nature of the active site means that structures that have been determined by crystallography or spectroscopy are likely to be protected, retained, and fully relevant in a PFE experiment. An electrocatalysis model formulated for the PFE of immobilized enzymes predicts interesting behavior and gives insight into why some hydrogenases are H2 producers and others are H2 oxidizers. Immobilization also allows for easy addition and removal of inhibitors along with precise potential control, one interesting outcome being that formaldehyde forms a reversible complex with reduced [FeFe]-hydrogenases, thereby providing insight into the order of electron and proton transfers. Experiments on O2-tolerant [NiFe]-hydrogenases show that O2 behaves like a reversible inhibitor: it

  20. Measuring interactions between polydimethylsiloxane and serum proteins at the air-water interface.

    PubMed

    Liao, Zhengzheng; Hsieh, Wan-Ting; Baumgart, Tobias; Dmochowski, Ivan J

    2013-07-30

    The interaction between synthetic polymers and proteins at interfaces is relevant to basic science as well as a wide range of applications in biotechnology and medicine. One particularly common and important interface is the air-water interface (AWI). Due to the special energetics and dynamics of molecules at the AWI, the interplay between synthetic polymer and protein can be very different from that in bulk solution. In this paper, we applied the Langmuir-Blodgett technique and fluorescence microscopy to investigate how the compression state of polydimethylsiloxane (PDMS) film at the AWI affects the subsequent adsorption of serum protein [e.g., human serum albumin (HSA) or immunoglobulin G (IgG)] and the interaction between PDMS and protein. Of particular note is our observation of circular PDMS domains with micrometer diameters that form at the AWI in the highly compressed state of the surface film: proteins were shown to adsorb preferentially to the surface of these circular PDMS domains, accompanied by a greater than 4-fold increase in protein found in the interfacial film. The PDMS-only film and the PDMS-IgG composite film were transferred to cover glass, and platinum-carbon replicas of the transferred films were further characterized by scanning electron microscopy and atomic force microscopy. We conclude that the structure of the PDMS film greatly affects the amount and distribution of protein at the interface.

  1. Structure of collagen adsorbed on a model implant surface resolved by polarization modulation infrared reflection-absorption spectroscopy

    NASA Astrophysics Data System (ADS)

    Brand, Izabella; Habecker, Florian; Ahlers, Michael; Klüner, Thorsten

    2015-03-01

    The polarization modulation infrared reflection-absorption spectra of collagen adsorbed on a titania surface and quantum chemical calculations are used to describe components of the amide I mode to the protein structure at a sub-molecular level. In this study, imino acid rich and poor fragments, representing the entire collagen molecule, are taken into account. The amide I mode of the collagen triple helix is composed of three absorption bands which involve: (i) (∼1690 cm-1) the Cdbnd O stretching modes at unhydrated groups, (ii) (1655-1673 cm-1) the Cdbnd O stretching at carbonyl groups at imino acids and glycine forming intramolecular hydrogen bonds with H atoms at both NH2 and, unusual for proteins, CH2 groups at glycine at a neighbouring chain and (iii) (∼1640 cm-1) the Cdbnd O stretching at carbonyl groups forming hydrogen bonds between two, often charged, amino acids as well as hydrogen bonds to water along the entire helix. The IR spectrum of films prepared from diluted solutions (c < 50 μg ml-1) corresponds to solution spectra indicating that native collagen molecules interact with water adsorbed on the titania surface. In films prepared from solutions (c ⩾ 50 μg ml-1) collagen multilayers are formed. The amide I mode is blue-shifted by 18 cm-1, indicating that intramolecular hydrogen bonds at imino acid rich fragments are weakened. Simultaneous red-shift of the amide A mode implies that the strength of hydrogen bonds at the imino acid poor fragments increases. Theoretically predicted distortion of the collagen structure upon adsorption on the titania surface is experimentally confirmed.

  2. Structure of collagen adsorbed on a model implant surface resolved by polarization modulation infrared reflection-absorption spectroscopy.

    PubMed

    Brand, Izabella; Habecker, Florian; Ahlers, Michael; Klüner, Thorsten

    2015-03-05

    The polarization modulation infrared reflection-absorption spectra of collagen adsorbed on a titania surface and quantum chemical calculations are used to describe components of the amide I mode to the protein structure at a sub-molecular level. In this study, imino acid rich and poor fragments, representing the entire collagen molecule, are taken into account. The amide I mode of the collagen triple helix is composed of three absorption bands which involve: (i) (∼1690cm(-1)) the CO stretching modes at unhydrated groups, (ii) (1655-1673cm(-1)) the CO stretching at carbonyl groups at imino acids and glycine forming intramolecular hydrogen bonds with H atoms at both NH2 and, unusual for proteins, CH2 groups at glycine at a neighbouring chain and (iii) (∼1640cm(-1)) the CO stretching at carbonyl groups forming hydrogen bonds between two, often charged, amino acids as well as hydrogen bonds to water along the entire helix. The IR spectrum of films prepared from diluted solutions (c<50μgml(-1)) corresponds to solution spectra indicating that native collagen molecules interact with water adsorbed on the titania surface. In films prepared from solutions (c⩾50μgml(-1)) collagen multilayers are formed. The amide I mode is blue-shifted by 18cm(-1), indicating that intramolecular hydrogen bonds at imino acid rich fragments are weakened. Simultaneous red-shift of the amide A mode implies that the strength of hydrogen bonds at the imino acid poor fragments increases. Theoretically predicted distortion of the collagen structure upon adsorption on the titania surface is experimentally confirmed.

  3. A novel fiber-based adsorbent technology

    SciTech Connect

    Reynolds, T.A.

    1997-10-01

    In this Phase I Small Business Innovation Research program, Chemica Technologies, Inc. is developing an economical, robust, fiber-based adsorbent technology for removal of heavy metals from contaminated water. The key innovation is the development of regenerable adsorbent fibers and adsorbent fiber cloths that have high capacity and selectivity for heavy metals and are chemically robust. The process has the potential for widespread use at DOE facilities, mining operations, and the chemical process industry.

  4. Molecular mobility in the monolayers of foam films stabilized by porcine lung surfactant.

    PubMed Central

    Lalchev, Z I; Todorov, R K; Christova, Y T; Wilde, P J; Mackie, A R; Clark, D C

    1996-01-01

    Certain physical properties of a range of foam film types that are believed to exist in vivo in the lung have been investigated. The contribution of different lung surfactant components found in porcine lung surfactant to molecular surface diffusion in the plane of foam films has been investigated for the first time. The influence of the type and thickness of black foam films, temperature, electrolyte concentration, and extract composition on surface diffusion has been studied using the fluorescence recovery after photobleaching technique. Fluorescent phospholipid probe molecules in foam films stabilized by porcine lung surfactant samples or their hydrophobic extracts consisting of surfactant lipids and hydrophobic lung surfactant proteins, SP-B and SP-C, exhibited more rapid diffusion than observed in films of its principal lipid component alone, L-alpha-phosphatidylcholine dipalmitoyl. This effect appears to be due to contributions from minor lipid components present in the total surfactant lipid extracts. The minor lipid components influence the surface diffusion in foam films both by their negative charge and by lowering the phase transition temperature of lung surfactant samples. In contrast, the presence of high concentrations of the hydrophillic surfactant protein A (SP-A) and non-lung-surfactant proteins in the sample reduced the diffusion coefficient (D) of the lipid analog in the adsorbed layer of the films. Hysteresis behavior of D was observed during temperature cycling, with the cooling curve lying above the heating curve. However, in cases where some surface molecular aggregation and surface heterogeneity were observed during cooling, the films became more rigid and molecules at the interfaces became immobilized. The thickness, size, capillary pressure, configuration, and composition of foam films of lung surfactant prepared in vitro support their investigation as realistic structural analogs of the surface films that exist in vivo in the lung

  5. Molecularly Imprinted Filtering Adsorbents for Odor Sensing

    PubMed Central

    Shinohara, Sho; Chiyomaru, You; Sassa, Fumihiro; Liu, Chuanjun; Hayashi, Kenshi

    2016-01-01

    Versatile odor sensors that can discriminate among huge numbers of environmental odorants are desired in many fields, including robotics, environmental monitoring, and food production. However, odor sensors comparable to an animal’s nose have not yet been developed. An animal’s olfactory system recognizes odor clusters with specific molecular properties and uses this combinatorial information in odor discrimination. This suggests that measurement and clustering of odor molecular properties (e.g., polarity, size) using an artificial sensor is a promising approach to odor sensing. Here, adsorbents composed of composite materials with molecular recognition properties were developed for odor sensing. The selectivity of the sensor depends on the adsorbent materials, so specific polymeric materials with particular solubility parameters were chosen to adsorb odorants with various properties. The adsorption properties of the adsorbents could be modified by mixing adsorbent materials. Moreover, a novel molecularly imprinted filtering adsorbent (MIFA), composed of an adsorbent substrate covered with a molecularly imprinted polymer (MIP) layer, was developed to improve the odor molecular recognition ability. The combination of the adsorbent and MIP layer provided a higher specificity toward target molecules. The MIFA thus provides a useful technique for the design and control of adsorbents with adsorption properties specific to particular odor molecules. PMID:27886070

  6. Capillary-channeled polymer (C-CP) films as processing platforms for protein analysis by matrix-assisted laser/desorption ionization mass spectrometry (MALDI-MS).

    PubMed

    Pittman, Jennifer J; Manard, Benjamin T; Kowalski, Paul J; Marcus, R Kenneth

    2012-01-01

    Polypropylene (PP) capillary-channeled polymer (C-CP) films have parallel, μm-sized channels that induce solution wicking via capillary action. Efficient mass transport from the solution phase to the channel surface leads to adsorption of hydrophobic protein solutes. The basic premise by which C-CP films can be used as media to manipulate analyte solutions (e.g., proteins in buffer), for the purpose of desalting or chromatographic separation prior to MALDI-MS analysis is presented here. Cytochrome c and myoglobin prepared in a Tris-HCl buffer, and ribonuclease A, lysozyme, and transferrin prepared in phosphate buffered saline (PBS), are used as the test solutions to demonstrate the desalting concept. Protein analysis is performed after deposition on a C-CP film with and without a water washing step, followed by spray deposition of a typical sinapinic acid matrix. Extracted MALDI mass spectra exhibit much improved signal-to-noise characteristics after water washing. A mixture of cytochrome c and myoglobin (2 μL of 2.5 μM each in Tris-HCl buffer) was applied, washed with water and spatially separated via simple capillary action (wicking) using a reversed-phase solvent composition of 0.1% trifluoroacetic acid (TFA) in 50:50 acetonitrile (ACN):H(2)O. Subsequent application of sinapinic acid followed by imaging of the film using MALDI-MS reveals that as the protein solution is wicked down the film, separation occurs.

  7. Capillary-Channeled Polymer (C-CP) Films as Processing Platforms for Protein Analysis by Matrix-Assisted Laser/Desorption Ionization Mass Spectrometry (MALDI-MS)

    NASA Astrophysics Data System (ADS)

    Pittman, Jennifer J.; Manard, Benjamin T.; Kowalski, Paul J.; Marcus, R. Kenneth

    2012-01-01

    Polypropylene (PP) capillary-channeled polymer (C-CP) films have parallel, μm-sized channels that induce solution wicking via capillary action. Efficient mass transport from the solution phase to the channel surface leads to adsorption of hydrophobic protein solutes. The basic premise by which C-CP films can be used as media to manipulate analyte solutions (e.g., proteins in buffer), for the purpose of desalting or chromatographic separation prior to MALDI-MS analysis is presented here. Cytochrome c and myoglobin prepared in a Tris-HCl buffer, and ribonuclease A, lysozyme, and transferrin prepared in phosphate buffered saline (PBS), are used as the test solutions to demonstrate the desalting concept. Protein analysis is performed after deposition on a C-CP film with and without a water washing step, followed by spray deposition of a typical sinapinic acid matrix. Extracted MALDI mass spectra exhibit much improved signal-to-noise characteristics after water washing. A mixture of cytochrome c and myoglobin (2 μL of 2.5 μM each in Tris-HCl buffer) was applied, washed with water and spatially separated via simple capillary action (wicking) using a reversed-phase solvent composition of 0.1% trifluoroacetic acid (TFA) in 50:50 acetonitrile (ACN):H2O. Subsequent application of sinapinic acid followed by imaging of the film using MALDI-MS reveals that as the protein solution is wicked down the film, separation occurs.

  8. Improved surface properties of polyaniline films by blending with Pluronic polymers without the modification of the other characteristics.

    PubMed

    Li, Z F; Ruckenstein, E

    2003-08-15

    Films of conductive polyaniline and amphiphilic Pluronic (P105) copolymer blends were prepared by dissolving the two polymers in N-methylpyrrolidinone (NMP) followed by a slow solvent evaporation at 55 degrees C. The characteristics of both doped and undoped films were determined by X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), water droplet contact angles, differential scanning calorimetry (DSC), thermal gravimetry analysis (TG), wide-angle X-ray diffraction (WAXD), and tensile strength measurements. The surface of the blends became more hydrophilic than that of the hydrophobic PANI film, but the other properties of the blends did not change appreciably for Pluronic content lower than 50 wt%. Compared to PANI films, the more hydrophilic surfaces decreased the amount of bovine serum albumin protein adsorbed. By preventing biofouling, the polyaniline-Pluronic blends can become more useful as biosensors than the polyaniline films.

  9. The uranium from seawater program at PNNL: Overview of marine testing, adsorbent characterization, adsorbent durability, adsorbent toxicity, and deployment studies

    DOE PAGES

    Gill, Gary A.; Kuo, Li -Jung; Janke, Christopher James; ...

    2016-02-07

    The Pacific Northwest National Laboratory's (PNNL) Marine Science Laboratory (MSL) located along the coast of Washington State is evaluating the performance of uranium adsorption materials being developed for seawater extraction under realistic marine conditions with natural seawater. Two types of exposure systems were employed in this program: flow-through columns for testing of fixed beds of individual fibers and pellets and a recirculating water flume for testing of braided adsorbent material. Testing consists of measurements of the adsorption of uranium and other elements from seawater as a function of time, typically 42 to 56 day exposures, to determine the adsorbent capacitymore » and adsorption rate (kinetics). Analysis of uranium and other trace elements collected by the adsorbents was conducted following strong acid digestion of the adsorbent with 50% aqua regia using either Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES) or Inductively Coupled Plasma Mass Spectrometer (ICP-MS). The ORNL 38H adsorbent had a 56 day adsorption capacity of 3.30 ± 0.68 g U/ kg adsorbent (normalized to a salinity of 35 psu), a saturation adsorption capacity of 4.89 ± 0.83 g U/kg of adsorbent material (normalized to a salinity of 35 psu) and a half-saturation time of 28 10 days. The AF1 adsorbent material had a 56 day adsorption capacity of 3.9 ± 0.2 g U/kg adsorbent material (normalized to a salinity of 35 psu), a saturation capacity of 5.4 ± 0.2 g U/kg adsorbent material (normalized to a salinity of 35 psu) and a half saturation time of 23 2 days. The ORNL amidoxime-based adsorbent materials are not specific for uranium, but also adsorb other elements from seawater. The major doubly charged cations in seawater (Ca and Mg) account for a majority of the cations adsorbed (61% by mass and 74% by molar percent). For the ORNL AF1 adsorbent material, U is the 4th most abundant element adsorbed by mass and 7th most abundant by molar percentage. Marine testing

  10. The uranium from seawater program at PNNL: Overview of marine testing, adsorbent characterization, adsorbent durability, adsorbent toxicity, and deployment studies

    SciTech Connect

    Gill, Gary A.; Kuo, Li -Jung; Janke, Christopher James; Park, Jiyeon; Jeters, Robert T.; Bonheyo, George T.; Pan, Horng -Bin; Wai, Chien; Khangaonkar, Tarang P.; Bianucci, Laura; Wood, Jordana R.; Warner, Marvin G.; Peterson, Sonja; Abrecht, David G.; Mayes, Richard T.; Tsouris, Costas; Oyola, Yatsandra; Strivens, Jonathan E.; Schlafer, Nicholas J.; Addleman, Shane R.; Chouyyok, Wilaiwan; Das, Sadananda; Kim, Jungseung; Buesseler, Ken; Breier, Crystal; D'Alessandro, Evan

    2016-02-07

    The Pacific Northwest National Laboratory's (PNNL) Marine Science Laboratory (MSL) located along the coast of Washington State is evaluating the performance of uranium adsorption materials being developed for seawater extraction under realistic marine conditions with natural seawater. Two types of exposure systems were employed in this program: flow-through columns for testing of fixed beds of individual fibers and pellets and a recirculating water flume for testing of braided adsorbent material. Testing consists of measurements of the adsorption of uranium and other elements from seawater as a function of time, typically 42 to 56 day exposures, to determine the adsorbent capacity and adsorption rate (kinetics). Analysis of uranium and other trace elements collected by the adsorbents was conducted following strong acid digestion of the adsorbent with 50% aqua regia using either Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES) or Inductively Coupled Plasma Mass Spectrometer (ICP-MS). The ORNL 38H adsorbent had a 56 day adsorption capacity of 3.30 ± 0.68 g U/ kg adsorbent (normalized to a salinity of 35 psu), a saturation adsorption capacity of 4.89 ± 0.83 g U/kg of adsorbent material (normalized to a salinity of 35 psu) and a half-saturation time of 28 10 days. The AF1 adsorbent material had a 56 day adsorption capacity of 3.9 ± 0.2 g U/kg adsorbent material (normalized to a salinity of 35 psu), a saturation capacity of 5.4 ± 0.2 g U/kg adsorbent material (normalized to a salinity of 35 psu) and a half saturation time of 23 2 days. The ORNL amidoxime-based adsorbent materials are not specific for uranium, but also adsorb other elements from seawater. The major doubly charged cations in seawater (Ca and Mg) account for a majority of the cations adsorbed (61% by mass and 74% by molar percent). For the ORNL AF1 adsorbent material, U is the 4th most abundant element adsorbed by mass and 7th most abundant by molar percentage. Marine testing at Woods

  11. Complete braided adsorbent for marine testing to demonstrate 3g-U/kg-adsorbent

    SciTech Connect

    Janke, Chris; Yatsandra, Oyola; Mayes, Richard; none,; Gill, Gary; Li-Jung, Kuo; Wood, Jordana; Sadananda, Das

    2014-04-30

    ORNL has manufactured four braided adsorbents that successfully demonstrated uranium adsorption capacities ranging from 3.0-3.6 g-U/kg-adsorbent in marine testing at PNNL. Four new braided and leno woven fabric adsorbents have also been prepared by ORNL and are currently undergoing marine testing at PNNL.

  12. Infrared Analysis Of Enzymes Adsorbed Onto Model Surfaces

    NASA Astrophysics Data System (ADS)

    Story, Gloria M.; Rauch, Deborah S.; Brode, Philip F.; Marcott, Curtis A.

    1989-12-01

    The adsorption of the enzymes, subtilisin BPN' and lysozyme, onto model surfaces was examined using attenuated total reflectance (ATR) infrared (IR) spectroscopy. Using a cylindrical internal reflection (CIRcle) cell with a Germanium (Ge) internal reflection element (IRE), model hydrophilic surfaces were made by plasma cleaning the IRE and model hydrophobic surfaces were made by precoating the IRE with a thin film of polystyrene. Gas chromatography (GC)-IR data collection software was used to monitor adsorption kinetics during the first five minutes after injection of the enzyme into the CIRcle cell. It was found that for both lysozyme and BPN', most of the enzyme that was going to adsorb onto the model surface did so within ten seconds after injection. Nearly an order-of-magnitude more BPN' adsorbed on the hydrophobic Ge surface than the hydrophilic one, while lysozyme adsorbed somewhat more strongly to the hydrophilic Ge surface. Overnight, the lysozyme layer continued to increase in thickness, while BPN' maintained its initial coverage. The appearance of carboxylate bands in some of the adsorbed BPN' spectra suggests the occurrence of peptide bond hydrolysis. A Au/Pd coating on the CIRcle cell o-rings had a significant effect on the adsorption of BPN'. (This coating was applied in an attempt to eliminate interfering Teflon absorption bands.) An apparent electrochemical reaction occurred, involving BPN', Ge, Au/Pd, and the salt solution used to stabilize BPN'. The result of this reaction was enhanced adsorption of the enzyme around the coated o-rings, etching of the Ge IRE at the o-ring site, and some autolysis of the enzyme. No such reaction was observed with lysozyme.

  13. Heterogeneous Reactions of Surface-Adsorbed Catechol: A Comparison of Tropospheric Aerosol Surrogates

    NASA Astrophysics Data System (ADS)

    Hinrichs, R. Z.; Woodill, L. A.

    2009-12-01

    reactivity of surface-adsorbed catechol contrasts prior observations which found thin films of pure catechol unreactive with NO2, indicating that thin films do not always serve as reliable models for surface-adsorbed species.

  14. Thin Film Research. Volume 1

    DTIC Science & Technology

    1985-05-30

    isotherm expected for nonwetting adsorbate-adsorbent systems. 8 Type I depicts monolayer adsorption. Type II is very common in the case of physical...5.11 show the isothermal growth rate as a function of relative humidity. The shape can be classified as a type IV adsorption isotherm that corresponds...by following the fringes generated by interference effects in the growing film. The Balzers 760 evaporation system was supplied with both types of

  15. Adsorbate Diffusion on Transition Metal Nanoparticles

    DTIC Science & Technology

    2015-01-01

    systematically studied adsorption and diffusion of atomic and diatomic species (H, C, N, O, CO, and NO) on nanometer-sized Pt and Cu nanoparticles with...species and two diatomic molecules (H, C, N, O, CO, and NO) as adsorbates and study the adsorption and diffusion of these adsorbates across the edges

  16. Langmuir-Blodgett films of cholesterol oxidase and S-layer proteins onto screen-printed electrodes

    NASA Astrophysics Data System (ADS)

    Guimarães, Juliana Aguilar; Ferraz, Helen Conceição; Alves, Tito Lívio Moitinho

    2014-04-01

    Stable Langmuir monolayers of cholesterol oxidase (ChOx) and S-layer proteins were produced at the water-air interface and subsequently transferred onto the surface of screen-printed carbon electrodes by the Langmuir-Blodgett (LB) technique. The modified electrode surface was characterized by atomic force microscopy (AFM) and cyclic voltammetry (CV). AFM indicated the presence of deposited layers, showing reduction of surface roughness (RMS and Rt parameters). Significant changes in the shape of CVs were observed in modified electrodes compared to bare electrodes. The anodic peaks could be observed in cyclic voltammograms (CV), at a scan rate equal to 25 mV s-1, using electrodes with Z-type LB deposition. The presence of S-layer proteins in the ChOx LB film increases the oxidation peak intensity and reduces the oxidation potential. Altogether, these results demonstrate the feasibility of producing a cholesterol biosensor based on the immobilization of ChOx and S-layer proteins by LB technique.

  17. Stable protein device platform based on pyridine dicarboxylic acid-bound cubic-nanostructured mesoporous titania films.

    PubMed

    Kim, Hwajeong; Park, Sung Soo; Seo, Jooyeok; Ha, Chang-Sik; Moon, Cheil; Kim, Youngkyoo

    2013-08-14

    Here we shortly report a protein device platform that is extremely stable in a buffer condition similar to human bodies. The protein device platform was fabricated by covalently attaching cytochrome c (cyt c) protein molecules to organic coupler molecules (pyridine dicarboxylic acid, PDA) that were already covalently bound to an electron-transporting substrate. A cubic nanostructured mesoporous titania film was chosen as an electron-transporting substrate because of its large-sized cubic holes (∼7 nm) and highly crystalline cubic titania walls (∼0.4 nm lattice). Binding of PDA molecules to the mesoporous titania surface was achieved by esterification reaction between carboxylic acid groups (PDA) and hydroxyl groups (titania) in the presence of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) mediator, whereas the immobilization of cyt c to the PDA coupler was carried out by the EDC-mediated amidation reaction between carboxylic acid groups (PDA) and amine groups (cyt c). Results showed that the 2,4-position isomer among several PDAs exhibited the highest oxidation and reduction peak currents. The cyt c-immobilized PDA-bound titania substrates showed stable and durable electrochemical performances upon continuous current-voltage cycling for 240 times (the final current change was less than 3%) and could detect superoxide that is a core indicator for various diseases including cancers.

  18. Database of Novel and Emerging Adsorbent Materials

    National Institute of Standards and Technology Data Gateway

    SRD 205 NIST/ARPA-E Database of Novel and Emerging Adsorbent Materials (Web, free access)   The NIST/ARPA-E Database of Novel and Emerging Adsorbent Materials is a free, web-based catalog of adsorbent materials and measured adsorption properties of numerous materials obtained from article entries from the scientific literature. Search fields for the database include adsorbent material, adsorbate gas, experimental conditions (pressure, temperature), and bibliographic information (author, title, journal), and results from queries are provided as a list of articles matching the search parameters. The database also contains adsorption isotherms digitized from the cataloged articles, which can be compared visually online in the web application or exported for offline analysis.

  19. NOx adsorber and method of regenerating same

    DOEpatents

    Endicott, Dennis L.; Verkiel, Maarten; Driscoll, James J.

    2007-01-30

    New technologies, such as NOx adsorber catalytic converters, are being used to meet increasingly stringent regulations on undesirable emissions, including NOx emissions. NOx adsorbers must be periodically regenerated, which requires an increased fuel consumption. The present disclosure includes a method of regenerating a NOx adsorber within a NOx adsorber catalytic converter. At least one sensor positioned downstream from the NOx adsorber senses, in the downstream exhaust, at least one of NOx, nitrous oxide and ammonia concentrations a plurality of times during a regeneration phase. The sensor is in communication with an electronic control module that includes a regeneration monitoring algorithm operable to end the regeneration phase when a time rate of change of the at least one of NOx, nitrous oxide and ammonia concentrations is after an expected plateau region begins.

  20. Picosecond adsorbate dynamics at condensed phase interfaces

    SciTech Connect

    Scott, T.W.; Chang, Y.J.; Martorell, J.

    1993-12-31

    Picosecond surface second harmonic generation has been used to probe a variety of elementary adsorbate reactions at liquid-solid interfaces. Electron transfer reactions at semiconductor-liquid junctions, geminate recombination of photogenerated free radical pairs and the orientational dynamics of dipolar adsorbates have all been explored in varying degrees of detail. These kinetic studies have led to a detailed analysis of adsorbate detection on the surface of non-centrosymmetric substrates as well as the use of total internal reflection geometries for signal enhancement from optically absorbing liquids. Particular emphasis has been placed on the static and dynamic characterization of adsorbate orientational distribution functions and how these are determined from the torque exerted on adsorbates by the angular part of the molecule-surface interaction potential.

  1. Fluorescence dynamics of microsphere-adsorbed sunscreens

    NASA Astrophysics Data System (ADS)

    Krishnan, R.

    2005-03-01

    Sunscreens are generally oily substances which are prepared in organic solvents, emulsions or dispersions with micro- or nanoparticles. These molecules adsorb to and integrate into skin cells. In order to understand the photophysical properties of the sunscreen, we compare steady-state and time-resolved fluorescence in organic solvent of varying dielectric constant ɛ and adsorbed to polystyrene microspheres and dispersed in water. Steady-state fluorescence is highest and average fluorescence lifetime longest in toluene, the solvent of lowest ɛ. However, there is no uniform dependence on ɛ. Sunscreens PABA and padimate-O show complex emission spectra. Microsphere-adsorbed sunscreens exhibit highly non-exponential decay, illustrative of multiple environments of the adsorbed molecule. The heterogeneous fluorescence dynamics likely characterizes sunscreen adsorbed to cells.

  2. Nanovalved Adsorbents for CH4 Storage.

    PubMed

    Song, Zhuonan; Nambo, Apolo; Tate, Kirby L; Bao, Ainan; Zhu, Minqi; Jasinski, Jacek B; Zhou, Shaojun J; Meyer, Howard S; Carreon, Moises A; Li, Shiguang; Yu, Miao

    2016-05-11

    A novel concept of utilizing nanoporous coatings as effective nanovalves on microporous adsorbents was developed for high capacity natural gas storage at low storage pressure. The work reported here for the first time presents the concept of nanovalved adsorbents capable of sealing high pressure CH4 inside the adsorbents and storing it at low pressure. Traditional natural gas storage tanks are thick and heavy, which makes them expensive to manufacture and highly energy-consuming to carry around. Our design uses unique adsorbent pellets with nanoscale pores surrounded by a coating that functions as a valve to help manage the pressure of the gas and facilitate more efficient storage and transportation. We expect this new concept will result in a lighter, more affordable product with increased storage capacity. The nanovalved adsorbent concept demonstrated here can be potentially extended for the storage of other important gas molecules targeted for diverse relevant functional applications.

  3. Titanium-silicon oxide film structures for polarization-modulated infrared reflection absorption spectroscopy

    PubMed Central

    Dunlop, Iain E.; Zorn, Stefan; Richter, Gunther; Srot, Vesna; Kelsch, Marion; van Aken, Peter A.; Skoda, Maximilian; Gerlach, Alexander; Spatz, Joachim P.; Schreiber, Frank

    2010-01-01

    We present a titanium-silicon oxide film structure that permits polarization modulated infrared reflection absorption spectroscopy on silicon oxide surfaces. The structure consists of a ~6 nm sputtered silicon oxide film on a ~200 nm sputtered titanium film. Characterization using conventional and scanning transmission electron microscopy, electron energy loss spectroscopy, X-ray photoelectron spectroscopy and X-ray reflectometry is presented. We demonstrate the use of this structure to investigate a selectively protein-resistant self-assembled monolayer (SAM) consisting of silane-anchored, biotin-terminated poly(ethylene glycol) (PEG). PEG-associated IR bands were observed. Measurements of protein-characteristic band intensities showed that this SAM adsorbed streptavidin whereas it repelled bovine serum albumin, as had been expected from its structure. PMID:20418963

  4. Surface modification of diamond-like carbon films with protein via polydopamine inspired coatings

    NASA Astrophysics Data System (ADS)

    Tao, Caihong; Yang, Shengrong; Zhang, Junyan; Wang, Jinqing

    2009-10-01

    In this paper, we report a facile two-step approach to immobilize proteins onto DLC surfaces. The first step was a simple immersion of DLC in a solution of dopamine. Polydopamine was deposited on DLC as a stable anchor to present protein molecules. Then the protein ad-layer was deposited on it. The chemical components of the modified DLC surfaces were characterized by Fourier transform infrared spectra and X-ray photoelectron spectroscopy. The biocompatibility of it was evaluated in vitro by the tetrazolium salt method. And it was indicated that the BSA modified surface had good haemocompatibility properties, and was cytocompatible to PC-12 cells.

  5. Micropatterned atmospheric pressure discharge surface modification of fluorinated polymer films for mammalian cell adhesion and protein binding

    NASA Astrophysics Data System (ADS)

    Graz, Ingrid; Ebner, Andreas; Bauer, Siegfried; Romanin, Christoph; Gruber, Hermann

    2008-08-01

    Micropatterning by an easily accessible atmospheric pressure discharge setup was performed on fluorinated polymer surfaces. Two conductively-coated glass slides are employed, together with the polymer foils to be modified and shadow masks for defining the microstructures. Surface angle measurements indicated the presence of charged groups in the surface of the fluoropolymer, enabling the growth of mammalian cells on arrays of spots with 600 μm diameter. XPS and FTIR spectra revealed the incorporation of oxygen in the surface, while the generation of aldehyde groups on the surface of fluorinated polymer films was demonstrated by selective coupling of fluorescence-labeled aminodextrane to the activated spots. The described method paves the way for producing protein microarray chips on flexible fluoropolymer substrates with standard laboratory equipment.

  6. Interfacial properties, thin film stability and foam stability of casein micelle dispersions.

    PubMed

    Chen, Min; Sala, Guido; Meinders, Marcel B J; van Valenberg, Hein J F; van der Linden, Erik; Sagis, Leonard M C

    2017-01-01

    Foam stability of casein micelle dispersions (CMDs) strongly depends on aggregate size. To elucidate the underlying mechanism, the role of interfacial and thin film properties was investigated. CMDs were prepared at 4°C and 20°C, designated as CMD4°C and CMD20°C. At equal protein concentrations, foam stability of CMD4°C (with casein micelle aggregates) was markedly higher than CMD20°C (without aggregates). Although the elastic modulus of CMD4°C was twice as that of CMD20°C at 0.005Hz, the protein adsorbed amount was slightly higher for CMD20°C than for CMD4°C, which indicated a slight difference in interfacial composition of the air/water interface. Non-linear surface dilatational rheology showed minor differences between mechanical properties of air/water interfaces stabilized by two CMDs. These differences in interfacial properties could not explain the large difference in foam stability between two CMDs. Thin film analysis showed that films made with CMD20°C drained to a more homogeneous film compared to films stabilized by CMD4°C. Large casein micelle aggregates trapped in the thin film of CMD4°C made the film more heterogeneous. The rupture time of thin films was significantly longer for CMD4°C (>1h) than for CMD20°C (<600s) at equal protein concentration. After homogenization, which broke down the aggregates, the thin films of CMD4°C became much more homogeneous, and both the rupture time of thin films and foam stability decreased significantly. In conclusion, the increased stability of foam prepared with CMD4°C appears to be the result of entrapment of casein micelle aggregates in the liquid films of the foam.

  7. Controlling Protein Oligomerization with Surface Curvature on the Nanoscale

    NASA Astrophysics Data System (ADS)

    Kurylowicz, Marty; Dutcher, John

    2011-03-01

    We investigate the effect of surface curvature on the conformation of beta-lactoglobulin (β LG) using Single Molecule Force Spectroscopy. β LG is a model interfacial protein which stabilizes oil droplets in milk and is known to undergo structural rearrangement when adsorbed onto a surface. We reliably control nanoscale surface curvature by creating close-packed monolayers of monodisperse polystyrene (PS) nanoparticles with diameters of 20, 40, 60, 80 and 140 nm, which are stable in aqueous buffer. By adsorbing β LG onto these hydrophobic surfaces and collecting force-extension curves in the fluid phase we can compare the conformation of β LG on 5 different surface curvatures with that on a flat PS film. We demonstrate a transition from oligomeric to monomeric β LG as the surface curvature is increased. Histograms of contour length from fits to peaks in the force-extension curves show a single maximum near 30 nm for β LG adsorbed onto nanoparticles with diameters less than 80 nm. For the larger nanoparticles, the histogram approaches that observed for β LG adsorbed onto a flat PS film, with maxima indicative of β LG dimers and trimers.

  8. Mechanical properties of hexadecane-water interfaces with adsorbed hydrophobic bacteria

    NASA Astrophysics Data System (ADS)

    Kang, Zhewen

    Certain strains of hydrophobic bacteria are known to play critical roles in petroleum-related applications. The aim of this study was to investigate how hydrophobic bacteria in their stationary phase could adsorb onto the hexadecane-water interface and alter its mechanical properties. The two strains of bacteria used in forming the interfacial films were Acinetobacter venetianus RAG-1 (a Gram-negative bacterium) and Rhodococcus erythropolis 20S-E1-c (Gram-positive). Experiments at two different length scales (millimetre and micrometre) were conducted and the results were compared. In addition, a simple flow experiment was designed in a constricted channel and the results were related to the intrinsic mechanical properties of bacteria-adsorbed films. On the millimetre scale, using the pendant drop technique, the film interfacial tension was monitored as the surface area was made to undergo changes. Under static conditions, both types of bacteria showed no significant effect on the interfacial tension. When subjected to transient excitations, the two bacterial films exhibited qualitatively similar, yet quantitative distinct rheological properties (including film elasticities and relaxation times). Under continuous reduction of surface area, the RAG-1 system showed a "paper-like" interface, while the interface of the 20S-E1-c system was "soap film-like." These macroscopic observations could be explained by the surface ultrastructures of the two cell strains. On the micrometre scale, using the micropipette technique, colloidal stability of the bacteria-coated oil droplets was examined through direct-contact experiments. Both types of bacteria were seen to function as effective stabilizers. In addition, the adsorbed bacteria also interacted with one another at the interface, giving rise to higher order 2-D rheological properties. A technique of directly probing the mechanical properties of the emulsion drop surfaces revealed that (a) the films behaved as purely elastic

  9. Low-Friction Adsorbed Layers of a Triblock Copolymer Additive in Oil-Based Lubrication.

    PubMed

    Yamada, Shinji; Fujihara, Ami; Yusa, Shin-ichi; Tanabe, Tadao; Kurihara, Kazue

    2015-11-10

    The tribological properties of the dilute solution of an ABA triblock copolymer, poly(11-acrylamidoundecanoic acid)-block-poly(stearyl methacrylate)-block-poly(11-acrylamidoundecanoic acid (A5S992A5), in poly(α-olefin) (PAO) confined between mica surfaces were investigated using the surface forces apparatus (SFA). Friction force was measured as a function of applied load and sliding velocity, and the film thickness and contact geometry during sliding were analyzed using the fringes of equal chromatic order (FECO) in the SFA. The results were contrasted with those of confined PAO films; the effects of the addition of A5S992A5 on the tribological properties were discussed. The thickness of the A5S992A5/PAO system varied with time after surface preparation and with repetitive sliding motions. The thickness was within the range from 40 to 70 nm 1 day after preparation (the Day1 film), and was about 20 nm on the following day (the Day2 film). The thickness of the confined PAO film was thinner than 1.4 nm, indicating that the A5S992A5/PAO system formed thick adsorbed layers on mica surfaces. The friction coefficient was about 0.03 to 0.04 for the Day1 film and well below 0.01 for the Day2 film, which were 1 or 2 orders of magnitude lower than the values for the confined PAO films. The time dependent changes of the adsorbed layer thickness and friction properties should be caused by the relatively low solubility of A5S992A5 in PAO. The detailed analysis of the contact geometry and friction behaviors implies that the particularly low friction of the Day2 film originates from the following factors: (i) shrinkage of the A5S992A5 molecules (mainly the poly(stearyl methacrylate) blocks) that leads to a viscoelastic properties of the adsorbed layers; and (ii) the intervening PAO layer between the adsorbed polymer layers that constitutes a high-fluidity sliding interface. Our results suggest that the block copolymer having relatively low solubility in a lubricant base oil is

  10. Inorganic chemically active adsorbents (ICAAs)

    SciTech Connect

    Ally, M.R.; Tavlarides, L.

    1997-10-01

    Oak Ridge National Laboratory (ORNL) researchers are developing a technology that combines metal chelation extraction technology and synthesis chemistry. They begin with a ceramic substrate such as alumina, titanium oxide or silica gel because they provide high surface area, high mechanical strength, and radiolytic stability. One preparation method involves silylation to hydrophobize the surface, followed by chemisorption of a suitable chelation agent using vapor deposition. Another route attaches newly designed chelating agents through covalent bonding by the use of coupling agents. These approaches provide stable and selective, inorganic chemically active adsorbents (ICAAs) tailored for removal of metals. The technology has the following advantages over ion exchange: (1) higher mechanical strength, (2) higher resistance to radiation fields, (3) higher selectivity for the desired metal ion, (4) no cation exchange, (5) reduced or no interference from accompanying anions, (6) faster kinetics, and (7) easy and selective regeneration. Target waste streams include metal-containing groundwater/process wastewater at ORNL`s Y-12 Plant (multiple metals), Savannah River Site (SRS), Rocky Flats (multiple metals), and Hanford; aqueous mixed wastes at Idaho National Engineering Laboratory (INEL); and scrubber water generated at SRS and INEL. Focus Areas that will benefit from this research include Mixed Waste, and Subsurface Contaminants.

  11. Fluorescence behavior of globular proteins from their bulk and thin film conformations in presence of mono-, di- and tri-valent ions.

    PubMed

    Bhowal, Ashim Chandra; Das, Kaushik; Kundu, Sarathi

    2015-09-01

    Photoluminescence behavior of globular proteins, lysozyme and bovine serum albumin (BSA), from their bulk and thin film conformations have been studied in presence of mono-, di- and tri-valent ions by using fluorescence and UV-Vis spectroscopy at two different temperatures and the morphology of the protein thin films have been studied by using atomic force microscopy. Protein- and ion-dependent dynamic and static quenching behaviors have been identified. While dynamic quenching is observed for lysozyme for all the three different valent ions, BSA shows no quenching for mono-valent (Na(+)) ions, dynamic quenching for di-valent (Ni(2+)) ions and static quenching for tri-valent (Fe(3+)) ions at pH≈5.5. After heat treatment, as the conformation of the protein molecules changes, the quenching efficiency for lysozyme in presence of ions decreases but shows enhancement for BSA. In thin film geometry, the molecular conformation of both lysozyme and BSA modifies on the solid surfaces and hence quenching efficiency also modifies in comparison with that of bulk and as a result the quenching efficiency for lysozyme increases but decreases for the BSA film.

  12. Effect of type and content of modified montmorillonite on the structure and properties of bio-nanocomposite films based on soy protein isolate and montmorillonite

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The non-biodegradable and non-renewable nature of plastic packaging has led to a renewed interest in packaging materials based on bio-nanocomposites (biopolymer matrix reinforced with nanoparticles such as layered silicates). Bio-nanocomposite films based on soy protein isolate (SPI) and modified mo...

  13. Plasma-induced graft-polymerization of polyethylene glycol acrylate on polypropylene films: chemical characterization and evaluation of the protein adsorption.

    PubMed

    Zanini, Stefano; Riccardi, Claudia; Grimoldi, Elisa; Colombo, Claudia; Villa, Anna Maria; Natalello, Antonino; Gatti-Lafranconi, Pietro; Lotti, Marina; Doglia, Silvia Maria

    2010-01-01

    This work deals with the optimization of argon plasma-induced graft-polymerization of polyethylene glycol acrylate (PEGA) on polypropylene (PP) films in order to obtain surfaces with a reduced protein adsorption for possible biomedical applications. To this end, we examined the protein adsorption on the treated and untreated surfaces. The graft-polymerization process consisted of four steps: (a) plasma pre-activation of the PP substrates; (b) immersion in a PEGA solution; (c) argon plasma-induced graft-polymerization; (d) washing and drying of the samples. The efficiency of these processes was evaluated in terms of the amount of grafted polymer, coverage uniformity and substrates wettability. The process was monitored by contact angle measurements, attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR), X-ray Photoelectron Spectroscopy (XPS) and atomic force microscopy (AFM) analyses. The stability of the obtained thin films was evaluated in water and in Phosphate Buffer Saline (PBS) at 37 degrees C. The adsorption of fibrinogen and green fluorescent protein (GFP)--taken as model proteins--on the differently prepared surfaces was evaluated through a fluorescence approach using laser scanning confocal microscopy with photon counting detection. After plasma treatments of short duration, the protein adsorption decreases by about 60-70% with respect to that of the untreated film, while long plasma exposure resulted in a higher protein adsorption, due to damaging of the grafted polymer.

  14. Fabrication of PP-g-PEGMA-g-heparin and its hemocompatibility: From protein adsorption to anticoagulant tendency

    NASA Astrophysics Data System (ADS)

    Jin, Jing; Jiang, Wei; shi, Qiang; Zhao, Jie; Yin, Jinghua; Stagnaro, Paola

    2012-05-01

    We described a two-step process to fabricate the heparinized polypropylene (PP) film using cyanuric chloride (CC) as a trifunctional reagent and poly (ethylene glycol) methacrylate (PEGMA) as a spacer. The modified PP films were characterized by attenuated total reflectance FT-IR and X-ray photoelectron spectroscopy; the content of PEGMA and heparin were determined by gravimetric method and a toluidine blue assay, respectively. For the PP-g-PEGMA films, it was found that small size protein BSA tended to adsorb on the surface of low molecular weight monomer grafted PP, whereas big spindle-shaped fibrinogen tended to adsorb on the surface of high molecular weight monomer grafted PP. We gave a definition of anti-protein adsorptive factor r with two model proteins, albumin and fibrinogen. The results by platelet adhesion and plasma recalcification time (PRT) experiments indicated that the factor r could be used to quantitatively evaluate the anticoagulant tendency of PP-g-PEGMA modified films. For the PP-g-PEGMA-g-heparin modified films, the surface was proved to have a high bioactivity by the adsorption of AT III assay and very low platelet adhesion. It indicated that immobilization of heparin on the PP film with PEGMA as a spacer was an effective way to improve the hemocompatibility of PP.

  15. Heat transfer to the adsorbent in solar adsorption cooling device

    NASA Astrophysics Data System (ADS)

    Pilat, Peter; Patsch, Marek; Papucik, Stefan; Vantuch, Martin

    2014-08-01

    The article deals with design and construction of solar adsorption cooling device and with heat transfer problem in adsorber. The most important part of adsorption cooling system is adsorber/desorber containing adsorbent. Zeolith (adsorbent) type was chosen for its high adsorption capacity, like a coolant was used water. In adsorber/desorber occur, at heating of adsorbent, to heat transfer from heat change medium to the adsorbent. The time required for heating of adsorber filling is very important, because on it depend flexibility of cooling system. Zeolith has a large thermal resistance, therefore it had to be adapted the design and construction of adsorber. As the best shows the tube type of adsorber with double coat construction. By this construction is ensured thin layer of adsorbent and heating is quick in all volume of adsorbent. The process of heat transfer was experimentally measured, but for comparison simulated in ANSYS, too.

  16. Adsorbed natural gas storage with activated carbon

    SciTech Connect

    Sun, Jian; Brady, T.A.; Rood, M.J.

    1996-12-31

    Despite technical advances to reduce air pollution emissions, motor vehicles still account for 30 to 70% emissions of all urban air pollutants. The Clean Air Act Amendments of 1990 require 100 cities in the United States to reduce the amount of their smog within 5 to 15 years. Hence, auto emissions, the major cause of smog, must be reduced 30 to 60% by 1998. Natural gas con be combusted with less pollutant emissions. Adsorbed natural gas (ANG) uses adsorbents and operates with a low storage pressure which results in lower capital costs and maintenance. This paper describes the production of an activated carbon adsorbent produced from an Illinois coal for ANG.

  17. Improvement of blood compatibility on polysulfone-polyvinylpyrrolidone blend films as a model membrane of dialyzer by physical adsorption of recombinant soluble human thrombomodulin (ART-123).

    PubMed

    Omichi, Masaaki; Matsusaki, Michiya; Maruyama, Ikuro; Akashi, Mitsuru

    2012-01-01

    ART-123 is a recombinant soluble human thrombomodulin (hTM) with potent anticoagulant activity, and is available for developing antithrombogenic surfaces by immobilization. We focused on improving blood compatibility on the dialyzer surface by the physical adsorption of ART-123 as a safe yet simple method without using chemical reagents. The physical adsorption mechanism and anticoagulant activities of adsorbed hTM on the surface of a polysulfone (PSF) membrane containing polyvinylpyrrolidone (PVP) as a model dialyzer were investigated in detail. The PVP content of the PSF-PVP films was saturated at 20 wt% after immersion in Tris-HCl buffer, even with the addition of over 20 wt% PVP. The surface morphology of the PSF-PVP films was strongly influenced by the PVP content, because PVP covered the outermost surface of the PSF-PVP films. The adsorption speed of hTM slowed dramatically with increasing PVP content up to 10 wt%, but the maximum adsorption amount of hTM onto the PSF-PVP film surface was almost the same, regardless of the PVP content. The PSF-PVP film with the physically adsorbed hTM showed higher protein C activity as compared to the PSF film, it showed excellent blood compatibility due to the protein C activity and the inhibition properties of platelet adhesion. The physical adsorption of hTM can be useful as a safe yet simple method to improve the blood compatibility of a dialyzer surface.

  18. Surface Acoustic Wave (SAW) Resonators for Monitoring Conditioning Film Formation.

    PubMed

    Hohmann, Siegfried; Kögel, Svea; Brunner, Yvonne; Schmieg, Barbara; Ewald, Christina; Kirschhöfer, Frank; Brenner-Weiß, Gerald; Länge, Kerstin

    2015-05-21

    We propose surface acoustic wave (SAW) resonators as a complementary tool for conditioning film monitoring. Conditioning films are formed by adsorption of inorganic and organic substances on a substrate the moment this substrate comes into contact with a liquid phase. In the case of implant insertion, for instance, initial protein adsorption is required to start wound healing, but it will also trigger immune reactions leading to inflammatory responses. The control of the initial protein adsorption would allow to promote the healing process and to suppress adverse immune reactions. Methods to investigate these adsorption processes are available, but it remains difficult to translate measurement results into actual protein binding events. Biosensor transducers allow user-friendly investigation of protein adsorption on different surfaces. The combination of several transduction principles leads to complementary results, allowing a more comprehensive characterization of the adsorbing layer. We introduce SAW resonators as a novel complementary tool for time-resolved conditioning film monitoring. SAW resonators were coated with polymers. The adsorption of the plasma proteins human serum albumin (HSA) and fibrinogen onto the polymer-coated surfaces were monitored. Frequency results were compared with quartz crystal microbalance (QCM) sensor measurements, which confirmed the suitability of the SAW resonators for this application.

  19. Surface Acoustic Wave (SAW) Resonators for Monitoring Conditioning Film Formation

    PubMed Central

    Hohmann, Siegfried; Kögel, Svea; Brunner, Yvonne; Schmieg, Barbara; Ewald, Christina; Kirschhöfer, Frank; Brenner-Weiß, Gerald; Länge, Kerstin

    2015-01-01

    We propose surface acoustic wave (SAW) resonators as a complementary tool for conditioning film monitoring. Conditioning films are formed by adsorption of inorganic and organic substances on a substrate the moment this substrate comes into contact with a liquid phase. In the case of implant insertion, for instance, initial protein adsorption is required to start wound healing, but it will also trigger immune reactions leading to inflammatory responses. The control of the initial protein adsorption would allow to promote the healing process and to suppress adverse immune reactions. Methods to investigate these adsorption processes are available, but it remains difficult to translate measurement results into actual protein binding events. Biosensor transducers allow user-friendly investigation of protein adsorption on different surfaces. The combination of several transduction principles leads to complementary results, allowing a more comprehensive characterization of the adsorbing layer. We introduce SAW resonators as a novel complementary tool for time-resolved conditioning film monitoring. SAW resonators were coated with polymers. The adsorption of the plasma proteins human serum albumin (HSA) and fibrinogen onto the polymer-coated surfaces were monitored. Frequency results were compared with quartz crystal microbalance (QCM) sensor measurements, which confirmed the suitability of the SAW resonators for this application. PMID:26007735

  20. Effects of thermally induced denaturation on technological-functional properties of whey protein isolate-based films.

    PubMed

    Schmid, M; Krimmel, B; Grupa, U; Noller, K

    2014-09-01

    This study examined how and to what extent the degree of denaturation affected the technological-functional properties of whey protein isolate (WPI)-based coatings. It was observed that denaturation affected the material properties of WPI-coated films significantly. Surface energy decreased by approximately 20% compared with native coatings. Because the surface energy of a coating should be lower than that of the substrate, this might result in enhanced wettability characteristics between WPI-based solution and substrate surface. Water vapor barrier properties increased by about 35% and oxygen barrier properties increased by approximately 33%. However, significant differences were mainly observed between coatings made of fully native WPI and ones with a degree of denaturation of 25%. Higher degrees of denaturation did not lead to further improvement of material properties. This observation offers cost-saving potential: a major share of denatured whey proteins may be replaced by fully native ones that are not exposed to energy-intensive heat treatment. Furthermore, native WPI solutions can be produced with higher dry matter content without gelatinizing. Hence, less moisture has to be removed through drying, resulting in reduced energy consumption.

  1. Pennycress protein isolate: Pilot plant production and application in films polymeric composites

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This work scaled up the process of producing pennycress protein isolates (PPI) using 5 kg starting material (previously 100 g in bench-scale research). Defatted press cake, produced by prepressing and hexane extraction, was mixed with preheated 50 L of aqueous NaOH (pH 10) for 90 min in a jacketed k...

  2. Unconventional superconductivity induced in Nb films by adsorbed chiral molecules

    NASA Astrophysics Data System (ADS)

    Alpern, H.; Katzir, E.; Yochelis, S.; Katz, N.; Paltiel, Y.; Millo, O.

    2016-11-01

    Motivated by recent observations of chiral-induced magnetization and spin-selective transport we studied the effect of chiral molecules on conventional BCS superconductors. By applying scanning tunneling spectroscopy, we demonstrate that the singlet-pairing s-wave order parameter of Nb is significantly altered upon adsorption of chiral polyalanine alpha-helix molecules on its surface. The tunneling spectra exhibit zero-bias conductance peaks embedded inside gaps or gap-like features, suggesting the emergence of unconventional triplet-pairing components with either d-wave or p-wave symmetry at the Nb organic-molecules interface, as corroborated by simulations. These results may open a way for realizing simple superconducting spintronics devices.

  3. Protein Adsorption in Three Dimensions

    PubMed Central

    Vogler, Erwin A.

    2011-01-01

    Recent experimental and theoretical work clarifying the physical chemistry of blood-protein adsorption from aqueous-buffer solution to various kinds of surfaces is reviewed and interpreted within the context of biomaterial applications, especially toward development of cardiovascular biomaterials. The importance of this subject in biomaterials surface science is emphasized by reducing the “protein-adsorption problem” to three core questions that require quantitative answer. An overview of the protein-adsorption literature identifies some of the sources of inconsistency among many investigators participating in more than five decades of focused research. A tutorial on the fundamental biophysical chemistry of protein adsorption sets the stage for a detailed discussion of the kinetics and thermodynamics of protein adsorption, including adsorption competition between two proteins for the same adsorbent immersed in a binary-protein mixture. Both kinetics and steady-state adsorption can be rationalized using a single interpretive paradigm asserting that protein molecules partition from solution into a three-dimensional (3D) interphase separating bulk solution from the physical-adsorbent surface. Adsorbed protein collects in one-or-more adsorbed layers, depending on protein size, solution concentration, and adsorbent surface energy (water wettability). The adsorption process begins with the hydration of an adsorbent surface brought into contact with an aqueous-protein solution. Surface hydration reactions instantaneously form a thin, pseudo-2D interface between the adsorbent and protein solution. Protein molecules rapidly diffuse into this newly-formed interface, creating a truly 3D interphase that inflates with arriving proteins and fills to capacity within milliseconds at mg/mL bulk-solution concentrations CB. This inflated interphase subsequently undergoes time-dependent (minutes-to-hours) decrease in volume VI by expulsion of either-or-both interphase water and

  4. X-ray fluorescence studies for the elemental composition and molecular organization of protein films on the surface of the liquid subphase

    SciTech Connect

    Zheludeva, S. I.; Novikova, N. N. Kovalchuk, M. V.; Stepina, N. D.; Konovalov, O. V.; Yurieva, E. A.

    2009-11-15

    This paper reports on the results of the investigation of protein films that are based on alkaline phosphatase and glucose oxidase enzymes and formed on the surface of the liquid subphase. The experimental studies have been performed using total external reflection X-ray fluorescence spectrometry at the European Synchrotron Radiation Facility (Grenoble, France). The self-organization processes that occur in protein systems on the surface of the liquid subphase under the conditions where the protein molecules retain their mobility have been investigated using X-ray fluorescence measurements for the first time.

  5. PERVAPORATION USING ADSORBENT-FILLED MEMBRANES

    EPA Science Inventory

    Membranes containing selective fillers, such as zeolites and activated carbon, can improve the separation by pervaporation. Applications of adsorbent-filled membranes in pervaporation have been demonstrated by a number of studies. These applications include removal of organic co...

  6. Chitin Adsorbents for Toxic Metals: A Review

    PubMed Central

    Anastopoulos, Ioannis; Bhatnagar, Amit; Bikiaris, Dimitrios N.; Kyzas, George Z.

    2017-01-01

    Wastewater treatment is still a critical issue all over the world. Among examined methods for the decontamination of wastewaters, adsorption is a promising, cheap, environmentally friendly and efficient procedure. There are various types of adsorbents that have been used to remove different pollutants such as agricultural waste, compost, nanomaterials, algae, etc., Chitin (poly-β-(1,4)-N-acetyl-d-glucosamine) is the second most abundant natural biopolymer and it has attracted scientific attention as an inexpensive adsorbent for toxic metals. This review article provides information about the use of chitin as an adsorbent. A list of chitin adsorbents with maximum adsorption capacity and the best isotherm and kinetic fitting models are provided. Moreover, thermodynamic studies, regeneration studies, the mechanism of adsorption and the experimental conditions are also discussed in depth. PMID:28067848

  7. Monitoring by Control Technique - Activated Carbon Adsorber

    EPA Pesticide Factsheets

    Stationary source emissions monitoring is required to demonstrate that a source is meeting the requirements in Federal or state rules. This page is about Activated Carbon Adsorber control techniques used to reduce pollutant emissions.

  8. IR investigations of surfaces and adsorbates

    SciTech Connect

    Gwyn Williams

    2001-12-10

    Synchrotron infrared reflection-absorption measurements on single crystal metal surfaces with adsorbates have led to the determination of many key parameters related to the bonding vibrational modes and the dynamics of adsorbates. In particular, energy couplings between electrons and adsorbate motion have been shown to be a dominant mechanism on metal surfaces. Excellent agreement has been obtained with calculations for many of the observations, and the synergy between theory and experiment has led to a deeper understanding of the roles of electrons and phonons in determining the properties of interfaces and their roles in phenomena as diverse as friction, lubrication, catalysis and adhesion. Nonetheless, as the experiments are pushed harder, to describe such effects as co-adsorbed systems, disagreements continue to challenge the theory and our comprehension also is still evolving.

  9. Bombyx mori silk protein films microprocessing with a nanosecond ultraviolet laser and a femtosecond laser workstation: theory and experiments

    NASA Astrophysics Data System (ADS)

    Lazare, S.; Sionkowska, A.; Zaborowicz, M.; Planecka, A.; Lopez, J.; Dijoux, M.; Louména, C.; Hernandez, M.-C.

    2012-01-01

    Laser microprocessing of several biopolymers from renewable resources is studied. Three proteinic materials were either extracted from the extracellular matrix like Silk Fibroin/Sericin and collagen, or coming from a commercial source like gelatin. All can find future applications in biomedical experimentation, in particular for cell scaffolding. Films of ˜hundred of microns thick were made by aqueous solution drying and laser irradiation. Attention is paid to the properties making them processable with two laser sources: the ultraviolet and nanosecond (ns) KrF (248 nm) excimer and the infrared and femtosecond (fs) Yb:KGW laser. The UV radiation is absorbed in a one-photon resonant process to yield ablation and the surface foaming characteristics of a laser-induced pressure wave. To the contrary, resonant absorption of the IR photons of the fs laser is not possible and does not take place. However, the high field of the intense I>˜1012 W/cm2 femtosecond laser pulse ionizes the film by the multiphoton absorption followed by the electron impact mechanism, yielding a dense plasma capable to further absorb the incident radiation of the end of the pulse. The theoretical model of this absorption is described in detail, and used to discuss the presented experimental effects (cutting, ablation and foaming) of the fs laser. The ultraviolet laser was used to perform simultaneous multiple spots experiments in which energetic foaming yields melt ejection and filament spinning. Airborne nanosize filaments "horizontally suspended by both ends" (0.25 μm diameter and 10 μm length) of silk biopolymer were observed upon irradiation with large fluences.

  10. Hydrophobic Porous Material Adsorbs Small Organic Molecules

    NASA Technical Reports Server (NTRS)

    Sharma, Pramod K.; Hickey, Gregory S.

    1994-01-01

    Composite molecular-sieve material has pore structure designed specifically for preferential adsorption of organic molecules for sizes ranging from 3 to 6 angstrom. Design based on principle that contaminant molecules become strongly bound to surface of adsorbent when size of contaminant molecules is nearly same as that of pores in adsorbent. Material used to remove small organic contaminant molecules from vacuum systems or from enclosed gaseous environments like closed-loop life-support systems.

  11. Regenerable activated bauxite adsorbent alkali monitor probe

    DOEpatents

    Lee, Sheldon H. D.

    1992-01-01

    A regenerable activated bauxite adsorber alkali monitor probe for field applications to provide reliable measurement of alkali-vapor concentration in combustion gas with special emphasis on pressurized fluidized-bed combustion (PFBC) off-gas. More particularly, the invention relates to the development of a easily regenerable bauxite adsorbent for use in a method to accurately determine the alkali-vapor content of PFBC exhaust gases.

  12. Regenerable activated bauxite adsorbent alkali monitor probe

    SciTech Connect

    Lee, S.H.D.

    1991-01-22

    This invention relates to a regenerable activated bauxite adsorber alkali monitor probe for field applications to provide reliable measurement of alkali-vapor 5 concentration in combustion gas with special emphasis on pressurized fluidized-bed combustion (PFBC) off-gas. More particularly, the invention relates to the development of a easily regenerable bauxite adsorbent for use in a method to accurately determine the alkali-vapor content of PFBC 10 exhaust gases.

  13. Regenerable activated bauxite adsorbent alkali monitor probe

    DOEpatents

    Lee, S.H.D.

    1992-12-22

    A regenerable activated bauxite adsorber alkali monitor probe for field applications to provide reliable measurement of alkali-vapor concentration in combustion gas with special emphasis on pressurized fluidized-bed combustion (PFBC) off-gas. More particularly, the invention relates to the development of a easily regenerable bauxite adsorbent for use in a method to accurately determine the alkali-vapor content of PFBC exhaust gases. 6 figs.

  14. Segregated ordered lipid phases and protein-promoted membrane cohesivity are required for pulmonary surfactant films to stabilize and protect the respiratory surface.

    PubMed

    Bernardino de la Serna, Jorge; Vargas, Rodolfo; Picardi, Victoria; Cruz, Antonio; Arranz, Rocío; Valpuesta, José M; Mateu, Leonardo; Pérez-Gil, Jesús

    2013-01-01

    Pulmonary surfactant is a lipid-protein complex essential to stabilize alveoli, by forming surface active films able to reach and sustain very low surface tensions (< 2 mN m(-1)) during the film compression that occurs at end-expiration. The particular lipid composition of surfactant, including a high proportion of dipalmitoylphosphatidylcholine (DPPC), induces segregation of fluid ordered and disordered phases in surfactant membranes and films at physiological temperatures. The segregation of DPPC-enriched ordered phase has been related with the ability of surfactant films to produce very low tensions, while the presence in surfactant of two specific hydrophobic polypeptides, SP-B and SP-C, is absolutely required to facilitate surfactant dynamics, including film formation and re-spreading during expansion at inspiration. In the present study, we have used X-ray scattering to analyze the structure of (1) whole native surfactant membranes purified from porcine lungs, (2) membranes reconstituted from the organic extract of surfactant containing the full lipid complement and the physiological proportion of SP-B and SP-C, and (3) membranes reconstituted from the lipid fraction of surfactant depleted of proteins. Small angle X-ray scattering data from whole surfactant or from membranes reconstituted from surfactant organic extract indicated the co-existence of two lamellar phases with different thicknesses. Such phase coexistence disappeared upon heating of the samples at temperatures above physiological values. When assessed in a captive bubble surfactometer, which mimics interfacial compression-expansion dynamics, the ability of surfactant films to produce very low tensions is only maintained at temperatures permitting the coexistence of the two lamellar phases. On the other hand, membranes reconstituted in the absence of proteins produced diffractograms indicative of the existence of a single dominant lamellar phase at all temperatures. These data suggest that SP

  15. Performance of a membrane adsorber for trace impurity removal in biotechnology manufacturing.

    PubMed

    Phillips, Michael; Cormier, Jason; Ferrence, Jennifer; Dowd, Chris; Kiss, Robert; Lutz, Herbert; Carter, Jeffrey

    2005-06-17

    Membrane adsorbers provide an attractive alternative to traditional bead-based chromatography columns used to remove trace impurities in downstream applications. A linearly scalable novel membrane adsorber family designed for the efficient removal of trace impurities from biotherapeutics, are capable of reproducibly achieving greater than 4 log removal of mammalian viruses, 3 log removal of endotoxin and DNA, and greater than 1 log removal of host cell protein. Single use, disposable membrane adsorbers eliminate the need for costly and time consuming column packing and cleaning validation associated with bead-based chromatography systems, and minimize the required number and volume of buffers. A membrane adsorber step reduces process time, floor space, buffer usage, labor cost, and improves manufacturing flexibility. This "process compression" effect is commonly associated with reducing the number of processing steps. The rigid microporous structure of the membrane layers allows for high process flux operation and uniform bed consistency at all processing scales.

  16. Microcalorimetric study of adsorption of glycomacropeptide on anion-exchange chromatography adsorbent.

    PubMed

    Lira, Rafael A; Minim, Luis A; Bonomo, Renata C F; Minim, Valéria P R; da Silva, Luis H M; da Silva, Maria C H

    2009-05-15

    The adsorption of glycomacropeptide (GMP) from cheese whey on an anion-exchange adsorbent was investigated using isothermal titration microcalorimetry to measure thermodynamic information regarding such processes. Isotherms data were measured at temperatures of 25 and 45 degrees C, pH 8.2 and various ionic strengths (0-0.08 molL(-1) NaCl). The equilibrium data were fit using the Langmuir model and the process was observed to be reversible. Temperature was observed to positively affect the interaction of the protein and adsorbent. Microcalorimetric studies indicated endothermic adsorption enthalpy in all cases, except at 45 degrees C and 0.0 molL(-1) NaCl. The adsorption process was observed to be entropically driven at all conditions studied. It was concluded that the increase in entropy, attributed to the release of hydration waters as well as bounded ions from the adsorbent and protein surface due to interactions of the protein and adsorbent, was a major driving force for the adsorption of GMP on the anion-exchange adsorbent. These results could allow for design of more effective ion-exchange separation processes for proteins.

  17. Immobilization of carboxymethylated polyethylenimine-metal-ion complexes in porous membranes to selectively capture his-tagged protein.

    PubMed

    Ning, Wenjing; Wijeratne, Salinda; Dong, Jinlan; Bruening, Merlin L

    2015-02-04

    Membrane adsorbers rapidly capture tagged proteins because flow through membrane pores efficiently conveys proteins to binding sites. Effective adsorbers, however, require membrane pores coated with thin films that bind multilayers of proteins. This work employs adsorption of polyelectrolytes that chelate metal ions to create functionalized membranes that selectively capture polyhistidine-tagged (His-tagged) proteins with binding capacities equal to those of high-binding commercial beads. Adsorption of functional polyelectrolytes is simpler than previous membrane-modification strategies such as growth of polymer brushes or derivatization of adsorbed layers with chelating moieties. Sequential adsorption of protonated poly(allylamine) (PAH) and carboxymethylated branched polyethylenimine (CMPEI) leads to membranes that bind Ni(2+) and capture ∼60 mg of His-tagged ubiquitin per mL of membrane. Moreover, these membranes enable isolation of His-tagged protein from cell lysates in <15 min. The backbone amine groups in CMPEI likely increase swelling in water to double protein binding compared to films composed of PAH and the chelating polymer poly[(N,N-dicarboxymethyl)allylamine] (PDCMAA), which has a hydrocarbon backbone. Metal leaching from PAH/CMPEI- and PAH/PDCMAA-modified membranes is similar to that from GE Hitrap FF columns. Eluates with 0.5 M imidazole contain <10 ppm of Ni(2+).

  18. Grafting of a model protein on lactide and caprolactone based biodegradable films for biomedical applications

    PubMed Central

    Larrañaga, Aitor; Guay-Bégin, Andrée-Anne; Chevallier, Pascale; Sabbatier, Gad; Fernández, Jorge; Laroche, Gaétan; Sarasua, Jose-Ramon

    2014-01-01

    Thermoplastic biodegradable polymers displaying elastomeric behavior and mechanical consistency are greatly appreciated for the regeneration of soft tissues and for various medical devices. However, while the selection of a suitable base material is determined by mechanical and biodegradation considerations, it is the surface properties of the biomaterial that are responsible for the biological response. In order to improve the interaction with cells and modulate their behavior, biologically active molecules can be incorporated onto the surface of the material. With this aim, the surface of a lactide and caprolactone based biodegradable elastomeric terpolymer was modified in two stages. First, the biodegradable polymer surface was aminated by atmospheric pressure plasma treatment and second a crosslinker was grafted in order to covalently bind the biomolecule. In this study, albumin was used as a model protein. According to X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM), albumin was efficiently immobilized on the surface of the terpolymer, the degree of albumin surface coverage (ΓBSA) reached ~35%. Moreover, gel permeation chromatography (GPC) studies showed that the hydrolytic degradation kinetic of the synthesized polymer was slightly delayed when albumin was grafted. However, the degradation process in the bulk of the material was unaffected, as demonstrated by Fourier transform infrared (FTIR) analyses. Furthermore, XPS analyses showed that the protein was still present on the surface after 28 days of degradation, meaning that the surface modification was stable, and that there had been enough time for the biological environment to interact with the modified material. PMID:24509417

  19. Insight into the adsorption of PPCPs by porous adsorbents: Effect of the properties of adsorbents and adsorbates.

    PubMed

    Zhu, Zengyin; Xie, Jiawen; Zhang, Mancheng; Zhou, Qing; Liu, Fuqiang

    2016-07-01

    Adsorption is an efficient method for removal of pharmaceuticals and personal care products (PPCPs). Magnetic resins are efficient adsorbents for water treatment and exhibit potential for PPCP removal. In this study, the magnetic hypercrosslinked resin Q100 was used for adsorption of PPCPs. The adsorption behavior of this resin was compared with those of two activated carbons, namely, Norit and F400D. Norit exhibited the fastest adsorption kinetics, followed by Q100. Norit featured a honeycomb shape and long-range ordered pore channels, which facilitated the diffusion of PPCPs. Moreover, the large average pore size of Q100 reduced diffusion resistance. The adsorbed amounts of 11 PPCPs on the three adsorbents increased with increasing adsorbate hydrophobicity. For Q100, a significant linear correlation was observed between the adsorption performance for PPCPs and hydrophobicity (logD value) of adsorbates (R(2) = 0.8951); as such, PPCPs with high logD values (>1.69) could be efficiently removed. Compared with those of Norit and F400D, the adsorption performance of Q100 was less affected by humic acid because of the dominant hydrophobic interaction. Furthermore, Q100 showed improved regeneration performance, which renders it promising for PPCP removal in practical applications.

  20. Controlling spins in adsorbed molecules by a chemical switch

    PubMed Central

    Wäckerlin, Christian; Chylarecka, Dorota; Kleibert, Armin; Müller, Kathrin; Iacovita, Cristian; Nolting, Frithjof; Jung, Thomas A.; Ballav, Nirmalya

    2010-01-01

    The development of chemical systems with switchable molecular spins could lead to the architecture of materials with controllable magnetic or spintronic properties. Here, we present conclusive evidence that the spin of an organometallic molecule coupled to a ferromagnetic substrate can be switched between magnetic off and on states by a chemical stimulus. This is achieved by nitric oxide (NO) functioning as an axial ligand of cobalt(II)tetraphenylporphyrin (CoTPP) ferromagnetically coupled to nickel thin-film (Ni(001)). On NO addition, the coordination sphere of Co2+ is modified and a NO–CoTPP nitrosyl complex is formed, which corresponds to an off state of the Co spin. Thermal dissociation of NO from the nitrosyl complex restores the on state of the Co spin. The NO-induced reversible off–on switching of surface-adsorbed molecular spins observed here is attributed to a spin trans effect. PMID:20975713

  1. Fabrication of Nanometer- and Micrometer-Scale Protein Structures by Site-Specific Immobilization of Histidine-Tagged Proteins to Aminosiloxane Films with Photoremovable Protein-Resistant Protecting Groups

    PubMed Central

    2016-01-01

    The site-specific immobilization of histidine-tagged proteins to patterns formed by far-field and near-field exposure of films of aminosilanes with protein-resistant photolabile protecting groups is demonstrated. After deprotection of the aminosilane, either through a mask or using a scanning near-field optical microscope, the amine terminal groups are derivatized first with glutaraldehyde and then with N-(5-amino-1-carboxypentyl)iminodiacetic acid to yield a nitrilo-triacetic-acid-terminated surface. After complexation with Ni2+, this surface binds histidine-tagged GFP and CpcA-PEB in a site-specific fashion. The chemistry is simple and reliable and leads to extensive surface functionalization. Bright fluorescence is observed in fluorescence microscopy images of micrometer- and nanometer-scale patterns. X-ray photoelectron spectroscopy is used to study quantitatively the efficiency of photodeprotection and the reactivity of the modified surfaces. The efficiency of the protein binding process is investigated quantitatively by ellipsometry and by fluorescence microscopy. We find that regions of the surface not exposed to UV light bind negligible amounts of His-tagged proteins, indicating that the oligo(ethylene glycol) adduct on the nitrophenyl protecting group confers excellent protein resistance; in contrast, exposed regions bind His-GFP very effectively, yielding strong fluorescence that is almost completely removed on treatment of the surface with imidazole, confirming a degree of site-specific binding in excess of 90%. This simple strategy offers a versatile generic route to the spatially selective site-specific immobilization of proteins at surfaces. PMID:26820378

  2. Using in situ X-ray reflectivity to study protein adsorption on hydrophilic and hydrophobic surfaces: benefits and limitations.

    PubMed

    Richter, Andrew G; Kuzmenko, Ivan

    2013-04-30

    We have employed in situ X-ray reflectivity (IXRR) to study the adsorption of a variety of proteins (lysozyme, cytochrome c, myoglobin, hemoglobin, serum albumin, and immunoglobulin G) on model hydrophilic (silicon oxide) and hydrophobic surfaces (octadecyltrichlorosilane self-assembled monolayers), evaluating this recently developed technique for its applicability in the area of biomolecular studies. We report herein the highest resolution depiction of adsorbed protein films, greatly improving on the precision of previous neutron reflectivity (NR) results and previous IXRR studies. We were able to perform complete scans in 5 min or less with the maximum momentum transfer of at least 0.52 Å(-1), allowing for some time-resolved information about the evolution of the protein film structure. The three smallest proteins (lysozyme, cytochrome c, and myoglobin) were seen to deposit as fully hydrated, nondenatured molecules onto hydrophilic surfaces, with indications of particular preferential orientations. Time evolution was observed for both lysozyme and myoglobin films. The larger proteins were not observed to deposit on the hydrophilic substrates, perhaps because of contrast limitations. On hydrophobic surfaces, all proteins were seen to denature extensively in a qualitatively similar way but with a rough trend that the larger proteins resulted in lower coverage. We have generated high-resolution electron density profiles of these denatured films, including capturing the growth of a lysozyme film. Because the solution interface of these denatured films is diffuse, IXRR cannot unambiguously determine the film extent and coverage, a drawback compared to NR. X-ray radiation damage was systematically evaluated, including the controlled exposure of protein films to high-intensity X-rays and exposure of the hydrophobic surface to X-rays before adsorption. Our analysis showed that standard measuring procedures used for XRR studies may lead to altered protein films

  3. Analysis of protein film voltammograms as Michaelis-Menten saturation curves yield the electron cooperativity number for deconvolution.

    PubMed

    Heering, Hendrik A

    2012-10-01

    Deconvolution of protein film voltammetric data by fitting multiple components (sigmoids, derivative peaks) often is ambiguous when features are partially overlapping, due to exchangeability between the width and the number of components. Here, a new method is presented to obtain the width of the components. This is based on the equivalence between the sigmoidal catalytic response as function of electrode potential, and the classical saturation curve obtained for the enzyme activity as function of the soluble substrate concentration, which is also sigmoidal when plotted versus log[S]. Thus, analysis of the catalytic voltammogram with Lineweaver-Burk, Eadie-Hofstee, and Hanes-Woolf plots is feasible. This provides a very sensitive measure of the cooperativity number (Hill coefficient), which for electrons equals the apparent (fractional) number of electrons that determine the width, and thereby the number of components (kinetic phases). This analysis is applied to the electrocatalytic oxygen reduction by Paracoccus denitrificans cytochrome aa(3) (cytochrome c oxidase). Four partially overlapping kinetic phases are observed that (stepwise) increase the catalytic efficiency with increasingly reductive potential. Translated to cell biology, the activity of the terminal oxidase stepwise adapts to metabolic demand for oxidative phosphorylation.

  4. Black Molecular Adsorber Coatings for Spaceflight Applications

    NASA Technical Reports Server (NTRS)

    Abraham, Nithin Susan; Hasegawa, Mark Makoto; Straka, Sharon A.

    2014-01-01

    The molecular adsorber coating is a new technology that was developed to mitigate the risk of on-orbit molecular contamination on spaceflight missions. The application of this coating would be ideal near highly sensitive, interior surfaces and instruments that are negatively impacted by outgassed molecules from materials, such as plastics, adhesives, lubricants, epoxies, and other similar compounds. This current, sprayable paint technology is comprised of inorganic white materials made from highly porous zeolite. In addition to good adhesion performance, thermal stability, and adsorptive capability, the molecular adsorber coating offers favorable thermal control characteristics. However, low reflectivity properties, which are typically offered by black thermal control coatings, are desired for some spaceflight applications. For example, black coatings are used on interior surfaces, in particular, on instrument baffles for optical stray light control. Similarly, they are also used within light paths between optical systems, such as telescopes, to absorb light. Recent efforts have been made to transform the white molecular adsorber coating into a black coating with similar adsorptive properties. This result is achieved by optimizing the current formulation with black pigments, while still maintaining its adsorption capability for outgassing control. Different binder to pigment ratios, coating thicknesses, and spray application techniques were explored to develop a black version of the molecular adsorber coating. During the development process, coating performance and adsorption characteristics were studied. The preliminary work performed on black molecular adsorber coatings thus far is very promising. Continued development and testing is necessary for its use on future contamination sensitive spaceflight missions.

  5. Size selective hydrophobic adsorbent for organic molecules

    NASA Technical Reports Server (NTRS)

    Sharma, Pramod K. (Inventor); Hickey, Gregory S. (Inventor)

    1997-01-01

    The present invention relates to an adsorbent formed by the pyrolysis of a hydrophobic silica with a pore size greater than 5 .ANG., such as SILICALITE.TM., with a molecular sieving polymer precursor such as polyfurfuryl alcohol, polyacrylonitrile, polyvinylidene chloride, phenol-formaldehyde resin, polyvinylidene difluoride and mixtures thereof. Polyfurfuryl alcohol is the most preferred. The adsorbent produced by the pyrolysis has a silicon to carbon mole ratio of between about 10:1 and 1:3, and preferably about 2:1 to 1:2, most preferably 1:1. The pyrolysis is performed as a ramped temperature program between about 100.degree. and 800.degree. C., and preferably between about 100.degree. and 600.degree. C. The present invention also relates to a method for selectively adsorbing organic molecules having a molecular size (mean molecular diameter) of between about 3 and 6 .ANG. comprising contacting a vapor containing the small organic molecules to be adsorbed with the adsorbent composition of the present invention.

  6. Membrane Perturbation Induced by Interfacially Adsorbed Peptides

    PubMed Central

    Zemel, Assaf; Ben-Shaul, Avinoam; May, Sylvio

    2004-01-01

    The structural and energetic characteristics of the interaction between interfacially adsorbed (partially inserted) α-helical, amphipathic peptides and the lipid bilayer substrate are studied using a molecular level theory of lipid chain packing in membranes. The peptides are modeled as “amphipathic cylinders” characterized by a well-defined polar angle. Assuming two-dimensional nematic order of the adsorbed peptides, the membrane perturbation free energy is evaluated using a cell-like model; the peptide axes are parallel to the membrane plane. The elastic and interfacial contributions to the perturbation free energy of the “peptide-dressed” membrane are evaluated as a function of: the peptide penetration depth into the bilayer's hydrophobic core, the membrane thickness, the polar angle, and the lipid/peptide ratio. The structural properties calculated include the shape and extent of the distorted (stretched and bent) lipid chains surrounding the adsorbed peptide, and their orientational (C-H) bond order parameter profiles. The changes in bond order parameters attendant upon peptide adsorption are in good agreement with magnetic resonance measurements. Also consistent with experiment, our model predicts that peptide adsorption results in membrane thinning. Our calculations reveal pronounced, membrane-mediated, attractive interactions between the adsorbed peptides, suggesting a possible mechanism for lateral aggregation of membrane-bound peptides. As a special case of interest, we have also investigated completely hydrophobic peptides, for which we find a strong energetic preference for the transmembrane (inserted) orientation over the horizontal (adsorbed) orientation. PMID:15189858

  7. Coating of peanuts with edible whey protein film containing alpha-tocopherol and ascorbyl palmitate.

    PubMed

    Han, J H; Hwang, H-M; Min, S; Krochta, J M

    2008-10-01

    Physical properties of whey protein isolate (WPI) coating solution incorporating ascorbic palmitate (AP) and alpha-tocopherol (tocopherol) were characterized, and the antioxidant activity of dried WPI coatings against lipid oxidation in roasted peanuts were investigated. The AP and tocopherol were mixed into a 10% (w/w) WPI solution containing 6.7% glycerol. Process 1 (P1) blended an AP and tocopherol mixture directly into the WPI solution using a high-speed homogenizer. Process 2 (P2) used ethanol as a solvent for dissolving AP and tocopherol into the WPI solution. The viscosity and turbidity of the WPI coating solution showed the Newtonian fluid behavior, and 0.25% of critical concentration of AP in WPI solution rheology. After peanuts were coated with WPI solutions, color changes of peanuts were measured during 16 wk of storage at 25 degrees C, and the oxidation of peanuts was determined by hexanal analysis using solid-phase micro-extraction samplers and GC-MS. Regardless of the presence of antioxidants in the coating layer, the formation of hexanal from the oxidation of peanut lipids was reduced by WPI coatings, which indicates WPI coatings protected the peanuts from oxygen permeation and oxidation. However, the incorporation of antioxidants in the WPI coating layer did not show a significant difference in hexanal production from that of WPI coating treatment without incorporation of antioxidants.

  8. Fluorescence properties of dyes adsorbed to silver islands, investigated by picosecond techniques

    NASA Astrophysics Data System (ADS)

    Leitner, A.; Lippitsch, M. E.; Draxler, S.; Riegler, M.; Aussenegg, F. R.

    1985-02-01

    The fluorescence properties of dye molecules (rhodamine 6G and erythrosin) adsorbed on pure glass surfaces and on silver islands films are investigated by cw and picosecond time-resolved methods. On pure glass surfaces we observe concentration quenching below a critical intermolecular distance (reduction of the fluorescence power per molecule as well as shortened and non-exponential fluorescence decay). On silver islands films the shortening in fluorescence lifetime is more drastic and is nearly independent of the intermolecular distance. This behavior suggests an electrodynamic interaction between dye monomers and plasmons in the metal particles, modified by a damping influence of dye dimers.

  9. Adsorbent catalytic nanoparticles and methods of using the same

    DOEpatents

    Slowing, Igor Ivan; Kandel, Kapil

    2017-01-31

    The present invention provides an adsorbent catalytic nanoparticle including a mesoporous silica nanoparticle having at least one adsorbent functional group bound thereto. The adsorbent catalytic nanoparticle also includes at least one catalytic material. In various embodiments, the present invention provides methods of using and making the adsorbent catalytic nanoparticles. In some examples, the adsorbent catalytic nanoparticles can be used to selectively remove fatty acids from feedstocks for biodiesel, and to hydrotreat the separated fatty acids.

  10. Mechanical properties of protein adsorption layers at the air/water and oil/water interface: a comparison in light of the thermodynamical stability of proteins.

    PubMed

    Mitropoulos, Varvara; Mütze, Annekathrin; Fischer, Peter

    2014-04-01

    Over the last decades numerous studies on the interfacial rheological response of protein adsorption layers have been published. The comparison of these studies and the retrieval of a common parameter to compare protein interfacial activity are hampered by the fact that different boundary conditions (e.g. physico-chemical, instrumental, interfacial) were used. In the present work we review previous studies and attempt a unifying approach for the comparison between bulk protein properties and their adsorption films. Among many common food grade proteins we chose bovine serum albumin, β-lactoglobulin and lysozyme for their difference in thermodynamic stability and studied their adsorption at the air/water and limonene/water interface. In order to achieve this we have i) systematically analyzed protein adsorption kinetics in terms of surface pressure rise using a drop profile analysis tensiometer and ii) we addressed the interfacial layer properties under shear stress using an interfacial shear rheometer under the same experimental conditions. We could show that thermodynamically less stable proteins adsorb generally faster and yield films with higher shear rheological properties at air/water interface. The same proteins showed an analog behavior when adsorbing at the limonene/water interface but at slower rates.

  11. Time Resolved Studies Of Adsorbed Species

    NASA Astrophysics Data System (ADS)

    Howard, J.; Nicol, J. M.

    1985-12-01

    A time-resolved Fourier transform IR study of ethyne adsorbed on ZnNaA zeolite yields results very different from those reported for related systems. Initially two species (A and B) are formed by the interaction of C2H2 with the cations. Whereas species A (π-bonded C2H2) was found to be removed immediately on evacuation, species B (probably Zn-acetylide) was not fully removed after 60 mins evacuation. In the presence of the gas phase, bands due to Species A decreased slowly in intensity as new bands due to adsorbed ethanal were observed.

  12. Incorporation of vitamin E in poly(3hydroxybutyrate)/Bioglass composite films: effect on surface properties and cell attachment.

    PubMed

    Misra, Superb K; Philip, Sheryl E; Chrzanowski, Wojciech; Nazhat, Showan N; Roy, Ipsita; Knowles, Jonathan C; Salih, Vehid; Boccaccini, Aldo R

    2009-04-06

    This study investigated the possibility of incorporating alpha-tocopherol (vitamin E) into poly(3hydroxybutyrate) (P(3HB))/Bioglass composites, which are being developed for bone tissue engineering matrices. P(3HB) films with 20 wt% Bioglass and 10 wt% vitamin E were prepared using the solvent casting technique. Addition of vitamin E significantly improved the hydrophilicity of the composites along with increasing the total protein adsorption. The presence of protein adsorbed on the composite surface was further confirmed using X-ray photoelectron spectroscopy analysis. Preliminary cell culture studies using MG-63 human osteoblasts showed that the addition of vitamin E in the P(3HB)/20 wt% Bioglass films significantly increased cell proliferation. The results achieved in this study confirmed the possibility of incorporating vitamin E as a suitable additive in P(3HB)/Bioglass composites to engineer the surface of the composites by promoting higher protein adsorption and increasing the hydrophilicity.

  13. Assembly of poly(dopamine)/poly(N-isopropylacrylamide) mixed films and their temperature-dependent interaction with proteins, liposomes, and cells.

    PubMed

    Zhang, Yan; Panneerselvam, Karthiga; Ogaki, Ryosuke; Hosta-Rigau, Leticia; van der Westen, Rebecca; Jensen, Bettina E B; Teo, Boon M; Zhu, Meifang; Städler, Brigitte

    2013-08-13

    Many biomedical applications benefit from responsive polymer coatings. The properties of poly(dopamine) (PDA) films can be affected by codepositing dopamine (DA) with the temperature-responsive polymer poly(N-isopropylacrylamide) (pNiPAAm). We characterize the film assembly at 24 and 39 °C using DA and aminated or carboxylated pNiPAAm by a quartz crystal microbalance with dissipation monitoring (QCM-D), X-ray photoelectron spectroscopy, UV-vis, ellipsometry, and atomic force microscopy. It was found that pNiPAAm with both types of end groups are incorporated into the films. We then identified a temperature-dependent adsorption behavior of proteins and liposomes to these PDA and pNiPAAm containing coatings by QCM-D and optical microscopy. Finally, a difference in myoblast cell response was found when these cells were allowed to adhere to these coatings. Taken together, these fundamental findings considerably broaden the potential biomedical applications of PDA films due to the added temperature responsiveness.

  14. A comparative study of fibrinogen adsorption onto metal oxide thin films

    NASA Astrophysics Data System (ADS)

    Silva-Bermudez, P.; Muhl, S.; Rodil, S. E.

    2013-10-01

    One of the first events occurring upon foreign material-biological medium contact is the adsorption of proteins, which evolution greatly determines the cells response to the material. Protein-surface interactions are a complex phenomenon driven by the physicochemical properties of the surface, protein(s) and liquid medium involve in the interaction. In this article the adsorption of fibrinogen (Fbg) onto Ta2O5, Nb2O5, TiO2 and ZrO2 thin films is reported. The adsorption kinetics and characteristics of the adsorbed fibrinogen layer were studied in situ using dynamic and spectroscopic ellipsometry. The films wettability, surface energy (γLW/AB) and roughness were characterized aiming to elucidate their correlations with Fbg adsorption. The adsorption rate changed accordingly to the film; the fastest adsorption rate and highest Fbg surface mass concentration (Γ) was observed on ZrO2. The hydrophobic/hydrophilic character of the oxide highly influenced Fbg adsorption. On Ta2O5, Nb2O5 and TiO2, which were either hydrophilic or in the breaking-point between hydrophilicity and hydrophobicity, Γ was correlated to the polar component of γLW/AB and roughness of the surface. On ZrO2, clearly hydrophobic, Γ increased significantly off the correlation observed for the other films. The results indicated different adsorption dynamics and orientations of the Fbg molecules dependent on the surface hydrophobic/hydrophilic character.

  15. Development and testing of molecular adsorber coatings

    NASA Astrophysics Data System (ADS)

    Abraham, Nithin S.; Hasegawa, Mark M.; Straka, Sharon A.

    2012-10-01

    The effect of on-orbit molecular contamination has the potential to degrade the performance of spaceflight hardware and diminish the lifetime of the spacecraft. For example, sensitive surfaces, such as optical surfaces, electronics, detectors, and thermal control surfaces, are vulnerable to the damaging effects of contamination from outgassed materials. The current solution to protect these surfaces is through the use of zeolite coated ceramic adsorber pucks. However, these pucks and its additional complex mounting hardware requirements result in several disadvantages, such as size, weight, and cost related concerns, that impact the spacecraft design and the integration and test schedule. As a result, a new innovative molecular adsorber coating was developed as a sprayable alternative to mitigate the risk of on-orbit molecular contamination. In this study, the formulation for molecular adsorber coatings was optimized using various binders, pigment treatment methods, binder to pigment ratios, thicknesses, and spray application techniques. The formulas that passed coating adhesion and vacuum thermal cycling were further tested for its adsorptive capacity. Accelerated molecular capacitance tests were performed in an innovatively designed multi-unit system containing idealized contaminant sources. This novel system significantly increased the productivity of the testing phase for the various formulations that were developed. Work performed during the development and testing phases has demonstrated successful application of molecular adsorber coatings onto metallic substrates, as well as, very promising results for the adhesion performance and the molecular capacitance of the coating. Continued testing will assist in the qualification of molecular adsorber coatings for use on future contamination sensitive spaceflight missions.

  16. Development and Testing of Molecular Adsorber Coatings

    NASA Technical Reports Server (NTRS)

    Abraham, Nithin; Hasegawa, Mark; Straka, Sharon

    2012-01-01

    The effect of on-orbit molecular contamination has the potential to degrade the performance of spaceflight hardware and diminish the lifetime of the spacecraft. For example, sensitive surfaces, such as optical surfaces, electronics, detectors, and thermal control surfaces, are vulnerable to the damaging effects of contamination from outgassed materials. The current solution to protect these surfaces is through the use of zeolite coated ceramic adsorber pucks. However, these pucks and its additional complex mounting hardware requirements result in several disadvantages, such as size, weight, and cost related concerns, that impact the spacecraft design and the integration and test schedule. As a result, a new innovative molecular adsorber coating was developed as a sprayable alternative to mitigate the risk of on-orbit molecular contamination. In this study, the formulation for molecular adsorber coatings was optimized using various binders, pigment treatment methods, binder to pigment ratios, thicknesses, and spray application techniques. The formulations that passed coating adhesion and vacuum thermal cycling tests were further tested for its adsorptive capacity. Accelerated molecular capacitance tests were performed in an innovatively designed multi-unit system containing idealized contaminant sources. This novel system significantly increased the productivity of the testing phase for the various formulations that were developed. Work performed during the development and testing phases has demonstrated successful application of molecular adsorber coatings onto metallic substrates, as well as, very promising results for the adhesion performance and the molecular capacitance of the coating. Continued testing will assist in the qualification of molecular adsorber coatings for use on future contamination sensitive spaceflight missions.

  17. Use of immobilized metal ions as a negative adsorbent for purification of enzymes: application to phosphoglycerate mutase from chicken muscle extract and horseradish peroxidase.

    PubMed

    Chaga, G; Andersson, L; Ersson, B; Berg, M

    1992-01-01

    Two enzymes, phosphoglycerate mutase and peroxidase, were purified by using an immobilized metal ion adsorbent for the removal of unwanted proteins. The mutase was obtained pure from a single column, whereas the purification of peroxidase required the use of a thiophilic adsorbent in a tandem. The capacity was 2.5 mg pure peroxidase per mL gel.

  18. Nanohardness, corrosion and protein adsorption properties of CuAlO2 films deposited on 316L stainless steel for biomedical applications

    NASA Astrophysics Data System (ADS)

    Chang, Shih-Hang; Chen, Jian-Zhang; Hsiao, Sou-Hui; Lin, Guan-Wei

    2014-01-01

    This study preliminarily assesses the biomedical applications of CuAlO2 coatings according to nanoindentation, electrochemical, and protein adsorption tests. Nanoindentation results revealed that the surface hardness of 316L stainless steel increased markedly after coating with CuAlO2 films. Electrochemical tests of corrosion potential, breakdown potential, and corrosion current density showed that the corrosion resistance properties of 316L stainless steel are considerably improved by CuAlO2 coatings. Bicinchoninic acid (BCA) protein assay results revealed that the protein adsorption behavior of 316L stainless steel did not exhibit notable differences with or without CuAlO2 coatings. A CuAlO2 coating of 100 nm thickness improved the surface nanohardness and corrosion resistance ability of 316L stainless steel. CuAlO2 is a potential candidate for biomaterial coating applications, particularly for surface modification of fine, delicate implants.

  19. Investigation of interparticle forces in natural waters: effects of adsorbed humic acids on iron oxide and alumina surface properties.

    PubMed

    Sander, Sylvia; Mosley, Luke M; Hunter, Keith A

    2004-09-15

    The nature of interparticle forces acting on colloid particle surfaces with adsorbed surface films of the internationally used humic acid standard material, Suwannee River Humic Acid (SHA), has been investigated using an atomic force microscope (AFM). Two particle surfaces were used, alumina and a hydrous iron oxide film coated onto silica particles. Adsorbed SHA dominated the interactive forces for both surface types when present. At low ionic strength and pH > 4, the force curves were dominated by electrostatic repulsion of the electrical double layers, with the extent of repulsion decreasing as electrolyte (NaCl) concentration increased, scaling with the Debye length (kappa(-1)) of the electrolyte according to classical theory. At pH approximately 4, electrostatic forces were largely absent, indicating almost complete protonation of carboxylic acid (-COOH) functional groups on the adsorbed SHA. Under these conditions and also at high electrolyte concentration ([NaCl] > 0.1 M), the absence of electrostatic forces allowed observation of repulsion forces arising from steric interaction of adsorbed SHA as the oxide surfaces approached closely to each other (separation < 10 nm). This steric barrier shrank as electrolyte concentration increased, implying tighter coiling of the adsorbed SHA molecules. In addition, adhesive bridging between surfaces was observed only in the presence of SHA films, implying a strong energy barrier to spontaneous detachment of the surfaces from each other once joined. This adhesion was especially strong in the presence of Ca2+ which appears to bridge SHA layers on each surface. Overall, our results show that SHA is a good model for the NOM adsorbed on colloids.

  20. Dynamic analysis of a closed-cycle solar adsorption refrigerator using two adsorbent-adsorbate pairs

    SciTech Connect

    Hajji, A. ); Worek, W. ); Lavan, Z. )

    1991-05-01

    In this paper a dynamic analysis of a closed-cycle, solar adsorption refrigerator is presented. The instantaneous and daily system performance are studied using two adsorbent-adsorbate pairs, Zeolite 13X-Water and Chabazite-Methanol. The effect of design and operating parameters, including inert material thermal capacitance, matrix porosity, and evaporation and condenser temperatures on the solar and cycle coefficients of performance are evaluated.

  1. Protein and polysaccharide-composite sol-gel silicate film for an interference-free amperometric glucose biosensor.

    PubMed

    Matsuhisa, Hironori; Tsuchiya, Munenori; Hasebe, Yasushi

    2013-11-01

    A novel permselective, organic-inorganic-hybrid, sol-gel silicate-film was chemically modified on an anodized platinum (Pt) electrode surface to form a selective, sensitive and interference-free amperometric glucose biosensor. This permselective hybrid sol-gel film consists of three organo-silanes [i.e., 3-aminopropyltriethoxysilane (APTES); tetraethoxysilane (TEOS); triethoxy-1H,1H,2H,2H-tridecafluoro-n-octylsilane (FAS)] and two biomacromolecules [i.e., bovine serum albumin (BSA) and a chitosan (CHIT)]. After the addition of the film to the Pt electrode, glucose oxidase (GOx) was covalently immobilized within the film with glutaraldehyde. The incorporation of the BSA and CHIT not only enhanced the permselectivity of H2O2 but also improved the activity of the immobilized GOx. The CHIT effectively suppressed any swelling of the film. Moreover, the conjugation of the FAS was especially effective in reducing the interference currents of AA and UA to levels less than 1/400 and 1/300 of the current of H2O2. The resulting organic-inorganic-hybrid sol-gel-film-based amperometric glucose biosensor exhibited rapid and sensitive responses to glucose (100% response in <3s, sensitivity: 1.84 μA mM(-1), detection limit: 0.032 mM), and the highly selective determination of glucose was possible, even in the presence of 0.1mM AA and UA.

  2. Characterization and protein-adsorption behavior of deposited organic thin film onto titanium by plasma polymerization with hexamethyldisiloxane.

    PubMed

    Hayakawa, Tohru; Yoshinari, Masao; Nemoto, Kimiya

    2004-01-01

    Plasma polymerized hexamethyldisiloxane (HMDSO) thin film was deposited onto titanium using a radio-frequency apparatus for the surface modification of titanium. A titanium disk was first polished using colloidal silica at pH=9.8. Plasma-polymerized HMDSO films were firmly attached to the titanium by heating the titanium to a temperature of approximately 250 degrees C. The thickness of the deposited film was 0.07-0.35mum after 10-60min of plasma polymerization. The contact angle with respect to double distilled water significantly increased after HMDSO coating. X-ray photoelectron spectroscopy revealed that the deposited thin film consisted of Si, C, and O atoms. No Ti peaks were observed on the deposited surface. The deposited HMDSO film was stable during 2-weeks immersion in phosphate buffer saline solution. Fourier transform reflection-absorption spectroscopy showed the formation of Si-H, Si-C, C-H, and Cz.dbnd6;O bonds in addition to Si-O-Si bonds. Quartz crystal microbalance-dissipation measurement demonstrated that the deposition of HMDSO thin films on titanium has a benefit for fibronectin adsorption at the early stage. In conclusion, plasma polymerization is a promising technique for the surface modification of titanium. HMDSO-coated titanium has potential application as a dental implant material.

  3. Thin films, asphaltenes, and reservoir wettability

    SciTech Connect

    Kaminsky, R.; Bergeron, V.; Radke, C.J. |

    1993-04-01

    Reservoir wettability impacts the success of oil recovery by waterflooding and other methods. To understand wettability and its alteration, thin-film forces in solid-aqueous-oil systems must be elucidated. Upon rupture of thick aqueous films separating the oil and rock phases, asphaltene components in the crude oil adsorb irreversibly on the solid surface, changing it from water-wet to oil-wet. Conditions of wettability alteration can be found by performing adhesion tests, in which an oil droplet is brought into contact with a solid surface. Exceeding a critical capillary pressure destabilizes the film, causing spontaneous film rupture to a molecularly adsorbed layer and oil adhesion accompanied by pinning at the three-phase contact line. The authors conduct adhesion experiments similar to those of Buckley and Morrow and simultaneously examine the state of the underlying thin film using optical microscopy and microinterferometry. Aqueous thin films between an asphaltic Orcutt crude oil and glass surfaces are studied as a function of aqueous pH and salinity. For the first time, they prove experimentally that strongly water-wet to strongly oil-wet wettability alteration and contact-angle pinning occur when thick aqueous films thin to molecularly adsorbed films and when the oil phase contains asphaltene molecules.

  4. EMERGING TECHNOLOGY SUMMARY: DEMONSTRATION OF AMBERSORB 563 ADSORBENT TECHNOLOGY

    EPA Science Inventory

    A field pilot study was conducted to demonstrate the technical feasibility and cost-effectiveness of Ambersorb® 5631 carbonaceous adsorbent for remediating groundwater contaminated with volatile organic compounds (VOCs). The Ambersorb adsorbent technology demonstration consist...

  5. Cleaning of biomaterial surfaces: protein removal by different solvents.

    PubMed

    Kratz, Fabian; Grass, Simone; Umanskaya, Natalia; Scheibe, Christian; Müller-Renno, Christine; Davoudi, Neda; Hannig, Matthias; Ziegler, Christiane

    2015-04-01

    The removal of biofilms or protein films from biomaterials is still a challenging task. In particular, for research investigations on real (applied) surfaces the reuse of samples is of high importance, because reuse allows the comparison of the same sample in different experiments. The aim of the present study was to evaluate the cleaning efficiency of different solvents (SDS, water, acetone, isopropanol, RIPA-buffer and Tween-20) on five different biomaterials (titanium, gold, PMMA (no acetone used), ceramic, and PTFE) with different wettability which were covered by layers of two different adsorbed proteins (BSA and lysozyme). The presence of a protein film after adsorption was confirmed by transmission electron microscopy (TEM). After treatment of the surfaces with the different solvents, the residual proteins on the surface were determined by BCA-assay (bicinchoninic acid assay). Data of the present study indicate that SDS is an effective solvent, but for several protein-substrate combinations it does not show the cleaning efficiency often mentioned in literature. RIPA-buffer and Tween-20 were more effective. They showed very low residual protein amounts after cleaning on all examined material surfaces and for both proteins, however, with small differences for the respective substrate-protein combinations. RIPA-buffer in combination with ultrasonication completely removed the protein layer as confirmed by TEM.

  6. Preparation of adsorbents for affinity chromatography using TSKgel Tresyl-Toyopearl 650M.

    PubMed

    Nakamura, K; Toyoda, K; Kato, Y; Shimura, K; Kasai, K

    1989-09-08

    The optimum conditions for the coupling of proteins were investigated using TSKgel Tresyl-Toyopearl 650M. They were dependent on the proteins coupled. For example, when soybean trypsin inhibitor was coupled at pH 8 the coupling was completed within 1 h and the subsequent adsorption capacity for trypsin was maximal. Longer coupling times decreased the adsorption capacity due to multi-point attachment. The adsorbents obtained were successfully used for affinity chromatography in a short time.

  7. An adsorbent monolith device to augment the removal of uraemic toxins during haemodialysis.

    PubMed

    Sandeman, Susan R; Howell, Carol A; Phillips, Gary J; Zheng, Yishan; Standen, Guy; Pletzenauer, Robert; Davenport, Andrew; Basnayake, Kolitha; Boyd, Owen; Holt, Stephen; Mikhalovsky, Sergey V

    2014-06-01

    Adsorbents designed with porosity which allows the removal of protein bound and high molecular weight uraemic toxins may improve the effectiveness of haemodialysis treatment of chronic kidney disease (CKD). A nanoporous activated carbon monolith prototype designed for direct blood contact was first assessed for its capacity to remove albumin bound marker toxins indoxyl sulphate (IS), p-cresyl sulphate (p-CS) and high molecular weight cytokine interleukin-6 in spiked healthy donor studies. Haemodialysis patient blood samples were then used to measure the presence of these markers in pre- and post-dialysis blood and their removal by adsorbent recirculation of post-dialysis blood samples. Nanopores (20-100 nm) were necessary for marker uraemic toxin removal during in vitro studies. Limited removal of IS and p-CS occurred during haemodialysis, whereas almost complete removal occurred following perfusion through the carbon monoliths suggesting a key role for such adsorbent therapies in CKD patient care.

  8. Analytical supercritical fluid extraction of adsorbent materials

    SciTech Connect

    Wright, B.W.; Wright, C.W.; Gale, R.W.; Smith, R.D.

    1987-01-01

    The use of supercritical fluids for the analytical extraction of semivolatile and higher molecular weight materials from various adsorbent and particulate matrices was investigated. Instrumentation was designed to allow gram quantities of the matrix to be extracted at pressures up to 400 bar and temperatures to 235 /sup 0/C with collection of the effluent in a sealed liquid-nitrogen-cooled flask. Carbon dioxide, isobutane, and methanol modified (20 mol %) carbon dioxide fluid systems were evaluated and compared to liquid Soxhlet extraction. Supercritical fluid extraction (SFE) provided very rapid (approx. =30 min) extraction with comparable efficiency to the Soxhlet methods, and both more rapid and more efficient extractions appear feasible. The more polar carbon dioxide-methanol fluid system gave higher extraction efficiencies for the more polar adsorbates and the isobutane system was more efficient for the higher molecular weight and less polar compounds.

  9. Efficient adsorbate transport on graphene by electromigration

    NASA Astrophysics Data System (ADS)

    Velizhanin, Kirill; Solenov, Dmitry

    2012-02-01

    Chemical functionalization of the surface of graphene holds promise for various applications ranging from nanoelectronics to surface catalysis and nano-assembling. In many practical situations it would be beneficial to be able to propel adsorbates along the graphene sheet in a controlled manner. We propose to use electromigration as an efficient means to transport adsorbates along the graphene surface. Within the tight-binding approximation for graphene, parametrized by density functional theory calculations, we estimate the contributions of the direct force and the electron wind force to the drift velocity of electromigration and demonstrate that the electromigration can be rather efficient. In particular, we show that the drift velocity of atomic oxygen covalently bound to graphene can reach up to 4 cm/s for realistic graphene samples. Further, we discuss ways to dynamically, i.e., during experiment, control the efficiency of electromigration by charging and/or local heating of graphene.

  10. Sand consolidation methods using adsorbable catalysts

    SciTech Connect

    Friedman, R. H.

    1985-04-23

    Methods are provided for selectively consolidating sand grains within a subterranean formation. First an acidic zirconium salt catalyst, such as ZrOCl/sub 2/, Zr(SO/sub 4/)/sub 2/, or ZrCl/sub 4/, is injected into the subterranean formation, wherein the acidic salt catalyst is adsorbed to the surface of the sand grains. Next a polymerizable resin composition such as furfuryl alcohol oligomer is introduced into the well formation. Polymerization of the resin occurs upon exposure to the elevated well temperatures and contact with the acid salt catalyst adsorbed to the sand grains. The polymerized resin serves to consolidate the surfaces of the sand grains while retaining permeability through the pore spaces. An ester of a weak organic acid is included with the resin compositions to control the extent of a polymerization by consuming the water by-product formed during the polymerization reaction.

  11. Gas storage using fullerene based adsorbents

    NASA Technical Reports Server (NTRS)

    Loutfy, Raouf O. (Inventor); Lu, Xiao-Chun (Inventor); Li, Weijiong (Inventor); Mikhael, Michael G. (Inventor)

    2000-01-01

    This invention is directed to the synthesis of high bulk density high gas absorption capacity adsorbents for gas storage applications. Specifically, this invention is concerned with novel gas absorbents with high gravimetric and volumetric gas adsorption capacities which are made from fullerene-based materials. By pressing fullerene powder into pellet form using a conventional press, then polymerizing it by subjecting the fullerene to high temperature and high inert gas pressure, the resulting fullerene-based materials have high bulk densities and high gas adsorption capacities. By pre-chemical modification or post-polymerization activation processes, the gas adsorption capacities of the fullerene-based adsorbents can be further enhanced. These materials are suitable for low pressure gas storage applications, such as oxygen storage for home oxygen therapy uses or on-board vehicle natural gas storage. They are also suitable for storing gases and vapors such as hydrogen, nitrogen, carbon dioxide, and water vapor.

  12. Analysis of Adsorbed Natural Gas Tank Technology

    NASA Astrophysics Data System (ADS)

    Knight, Ernest; Schultz, Conrad; Rash, Tyler; Dohnke, Elmar; Stalla, David; Gillespie, Andrew; Sweany, Mark; Seydel, Florian; Pfeifer, Peter

    With gasoline being an ever decreasing finite resource and with the desire to reduce humanity's carbon footprint, there has been an increasing focus on innovation of alternative fuel sources. Natural gas burns cleaner, is more abundant, and conforms to modern engines. However, storing compressed natural gas (CNG) requires large, heavy gas cylinders, which limits space and fuel efficiency. Adsorbed natural gas (ANG) technology allows for much greater fuel storage capacity and the ability to store the gas at a much lower pressure. Thus, ANG tanks are much more flexible in terms of their size, shape, and weight. Our ANG tank employs monolithic nanoporous activated carbon as its adsorbent material. Several different configurations of this Flat Panel Tank Assembly (FPTA) along with a Fuel Extraction System (FES) were examined to compare with the mass flow rate demands of an engine.

  13. Phase transitions in films of lung surfactant at the air-water interface.

    PubMed Central

    Nag, K; Perez-Gil, J; Ruano, M L; Worthman, L A; Stewart, J; Casals, C; Keough, K M

    1998-01-01

    45 mN/m. It also induced formation of large amounts of novel, nearly circular domains containing probe above pi of 50 mN/m, these domains being different in appearance than any seen at lower pressures with calcium or higher pressures in the absence of calcium. Surfactant protein-A (SP-A) adsorbed from the subphase onto solvent-spread LSE films, and aggregated condensed domains in presence of calcium. This study indicates that spread or adsorbed lung surfactant films can undergo expanded to condensed, and possibly other, phase transitions at the air-water interface as lateral packing density increases. These phase transitions are affected by divalent cations and SP-A in the subphase, and possibly by loss of material from the surface upon cyclic compression and expansion. PMID:9635752

  14. Remediation of AMD using industrial waste adsorbents

    NASA Astrophysics Data System (ADS)

    Mohammed, Nuur Hani Bte; Yaacob, Wan Zuhairi Wan

    2016-11-01

    The study investigates the characteristic of industrial waste as adsorbents and its potential as heavy metals absorbents in AMD samples. The AMD sample was collected from active mine pond and the pH was measured in situ. The metal contents were analyzed by ICP-MS. The AMD water was very acidic (pH< 3.5), and the average heavy metals content in AMD were high especially in Fe (822.029 mg/l). Fly ash was found to be the most effective absorbent material containing high percentage of CaO (57.24%) and SiO2 (13.88%), followed by ladle furnace slag containing of high amount of CaO (51.52%) and Al2O3 (21.23%), while biomass ash consists of SiO2 (43.07%) and CaO (12.97%). Tank analysis display a huge changes due to pH value change from acidity to nearly neutral phases. After 50 days, fly ash remediation successfully increase the AMD pH values from pH 2.57-7.09, while slag change from acidity to nearly alkaline phase from pH 2.60-7.3 and biomass has change to pH 2.54-6.8. Fly ash has successfully remove Fe, Mn, Cu, and Ni. Meanwhile, slag sample displays as an effective adsorbent to adsorb more Pb and Cd in acid mine drainage.

  15. Local anesthetics adsorbed onto infusion balloon.

    PubMed

    Mizogami, Maki; Tsuchiya, Hironori; Takakura, Ko

    2004-09-01

    We compared the adsorption of different local anesthetics onto infusion balloons and studied one of the possible mechanisms for adsorption. After injection of lidocaine, bupivacaine, ropivacaine, and mepivacaine solutions (1 mM each; pH 7.4) into balloons of 100-mL volume, their concentrations in effluents flowing out at 4 mL/h were determined over time by high-performance liquid chromatography. All were adsorbed in a structure-dependent manner, and the concentration decreased by 6%-14% within 5 min. Bupivacaine was most strongly adsorbed, followed by lidocaine, ropivacaine, and mepivacaine. QX-314, a quaternary ammonium derivative of lidocaine, was only weakly adsorbed compared with the parent compound lidocaine. The extent of adsorption of local anesthetics was related to their hydrophobicity (evaluated by reversed-phase chromatography) and was much more at pH 7.4 than at pH 6.0. A hydrophobic interaction with balloon materials appears to be responsible for the adsorption of local anesthetics. When infusion balloons are used for the continuous administration of local anesthetics, attention should be paid to the possibility that their actual concentrations in effluents are smaller than those present when they are initially prepared.

  16. Adsorbed Natural Gas Storage in Optimized High Surface Area Microporous Carbon

    NASA Astrophysics Data System (ADS)

    Romanos, Jimmy; Rash, Tyler; Nordwald, Erik; Shocklee, Joshua Shawn; Wexler, Carlos; Pfeifer, Peter

    2011-03-01

    Adsorbed natural gas (ANG) is an attractive alternative technology to compressed natural gas (CNG) or liquefied natural gas (LNG) for the efficient storage of natural gas, in particular for vehicular applications. In adsorbants engineered to have pores of a few molecular diameters, a strong van der Walls force allows reversible physisorption of methane at low pressures and room temperature. Activated carbons were optimized for storage by varying KOH:C ratio and activation temperature. We also consider the effect of mechanical compression of powders to further enhance the volumetric storage capacity. We will present standard porous material characterization (BET surface area and pore-size distribution from subcritical N2 adsorption) and methane isotherms up to 250 bar at 293K. At sufficiently high pressure, specific surface area, methane binding energy and film density can be extracted from supercritical methane adsorption isotherms. Research supported by the California Energy Commission (500-08-022).

  17. Energy of adhesion of human T cells to adsorption layers of monoclonal antibodies measured by a film trapping technique.

    PubMed Central

    Ivanov, I B; Hadjiiski, A; Denkov, N D; Gurkov, T D; Kralchevsky, P A; Koyasu, S

    1998-01-01

    A novel method for studying the interaction of biological cells with interfaces (e.g., adsorption monolayers of antibodies) is developed. The method is called the film trapping technique because the cell is trapped within an aqueous film of equilibrium thickness smaller than the cell diameter. A liquid film of uneven thickness is formed around the trapped cell. When observed in reflected monochromatic light, this film exhibits an interference pattern of concentric bright and dark fringes. From the radii of the fringes one can restore the shape of interfaces and the cell. Furthermore, one can calculate the adhesive energy between the cell membrane and the aqueous film surface (which is covered by a layer of adsorbed proteins and/or specific ligands), as well as the disjoining pressure, representing the force of interaction per unit area of the latter film. The method is applied to two human T cell lines: Jurkat and its T cell receptor negative (TCR-) derivative. The interaction of these cells with monolayers of three different monoclonal antibodies adsorbed at a water-air interface is studied. The results show that the adhesive energy is considerable (above 0.5 mJ/m2) when the adsorption monolayer contains antibodies acting as specific ligands for the receptors expressed on the cell surface. In contrast, the adhesive energy is close to zero in the absence of such a specific ligand-receptor interaction. In principle, the method can be applied to the study of the interaction of a variety of biological cells (B cells, natural killer cells, red blood cells, etc.) with adsorption monolayers of various biologically active molecules. In particular, film trapping provides a tool for the gentle micromanipulation of cells and for monitoring of processes (say the activation of a T lymphocyte) occurring at the single-cell level. PMID:9649417

  18. The effect of nanoscale surface curvature on the oligomerization of surface-bound proteins

    PubMed Central

    Kurylowicz, M.; Paulin, H.; Mogyoros, J.; Giuliani, M.; Dutcher, J. R.

    2014-01-01

    The influence of surface topography on protein conformation and association is used routinely in biological cells to orchestrate and coordinate biomolecular events. In the laboratory, controlling the surface curvature at the nanoscale offers new possibilities for manipulating protein–protein interactions and protein function at surfaces. We have studied the effect of surface curvature on the association of two proteins, α-lactalbumin (α-LA) and β-lactoglobulin (β-LG), which perform their function at the oil–water interface in milk emulsions. To control the surface curvature at the nanoscale, we have used a combination of polystyrene (PS) nanoparticles (NPs) and ultrathin PS films to fabricate chemically pure, hydrophobic surfaces that are highly curved and are stable in aqueous buffer. We have used single-molecule force spectroscopy to measure the contour lengths Lc for α-LA and β-LG adsorbed on highly curved PS surfaces (NP diameters of 27 and 50 nm, capped with a 10 nm thick PS film), and we have compared these values in situ with those measured for the same proteins adsorbed onto flat PS surfaces in the same samples. The Lc distributions for β-LG adsorbed onto a flat PS surface contain monomer and dimer peaks at 60 and 120 nm, respectively, while α-LA contains a large monomer peak near 50 nm and a dimer peak at 100 nm, with a tail extending out to 200 nm, corresponding to higher order oligomers, e.g. trimers and tetramers. When β-LG or α-LA is adsorbed onto the most highly curved surfaces, both monomer peaks are shifted to much smaller values of Lc. Furthermore, for β-LG, the dimer peak is strongly suppressed on the highly curved surface, whereas for α-LA the trimer and tetramer tail is suppressed with no significant change in the dimer peak. For both proteins, the number of higher order oligomers is significantly reduced as the curvature of the underlying surface is increased. These results suggest that the surface curvature provides a new

  19. Formation Process of Eosin Y-Adsorbing ZnO Particles by Electroless Deposition and Their Photoelectric Conversion Properties.

    PubMed

    Nagaya, Satoshi; Nishikiori, Hiromasa; Mizusaki, Hideaki; Wagata, Hajime; Teshima, Katsuya

    2015-06-03

    The thin films consisting of crystalline ZnO particles were prepared on fluorine-doped tin oxide electrodes by electroless deposition. The particles were deposited from an aqueous solution containing zinc nitrate, dimethyamine-borane, and eosin Y at 328 K. As the Pd particles were adsorbed on the substrate, not only the eosin Y monomer but also the dimer and debrominated species were rapidly adsorbed on the spherical ZnO particles, which were aggregated and formed secondary particles. On the other hand, in the absence of the Pd particles, the monomer was adsorbed on the flake-shaped ZnO particles, which vertically grew on the substrate surface and had a high crystallinity. The photoelectric conversion efficiency was higher for the ZnO electrodes containing a higher amount of the monomer during light irradiation.

  20. A Method To Measure Protein Unfolding at an Air-Liquid Interface.

    PubMed

    Leiske, Danielle L; Shieh, Ian C; Tse, Martha Lovato

    2016-10-04

    Proteins are surface-active molecules that have a propensity to adsorb to hydrophobic interfaces, such as the air-liquid interface. Surface flow can increase aggregation of adsorbed proteins, which may be an undesirable consequence depending on the application. As changes in protein conformation upon adsorption are thought to induce aggregation, the ability to measure the folded state of proteins at interfaces is of particular interest. However, few techniques currently exist to measure protein conformation at interfaces. Here we describe a technique capable of measuring the hydrophobicity, and therefore the conformation and folded state, of proteins at air-liquid interfaces by exploiting the environmentally sensitive fluorophore Nile red. Two monoclonal antibodies (mAbs) with high (mAb1) and low (mAb2) surface activity were used to highlight the technique. Both mAbs showed low background fluorescence of Nile red in the liquid subphase and at a glass-liquid interface. In contrast, at the air-liquid interface Nile red fluorescence for mAb1 increased immediately after protein adsorption, whereas the Nile red fluorescence of the mAb2 film evolved more slowly in time even though the adsorbed quantity of protein remained constant. The results demonstrate that hydrophobicity upon mAb adsorption to the air-liquid interface evolves in a time-dependent manner. Interfacial hydrophobicity may be indicative of protein conformation or folded state, where rapid unfolding of mAb1 upon adsorption would be consistent with increased protein aggregation compared to mAb2. The ability to measure protein hydrophobicity at interfaces using Nile red, combined with small sample requirements and minimal sample preparation, fills a gap in existing interfacial techniques.

  1. Modification and patterning of nanometer-thin poly(ethylene glycol) films by electron irradiation.

    PubMed

    Meyerbröker, Nikolaus; Zharnikov, Michael

    2013-06-12

    In this study, we analyzed the effect of electron irradiation on highly cross-linked and nanometer-thin poly(ethylene glycol) (PEG) films and, in combination with electron beam lithography (EBL), tested the possibility to prepare different patterns on their basis. Using several complementary spectroscopic techniques, we demonstrated that electron irradiation results in significant chemical modification and partial desorption of the PEG material. The initially well-defined films were progressively transformed in carbon-enriched and oxygen-depleted aliphatic layers with, presumably, still a high percentage of intermolecular cross-linking bonds. The modification of the films occurred very rapidly at low doses, slowed down at moderate doses, and exhibited a leveling off behavior at higher doses. On the basis of these results, we demonstrated the fabrication of wettability patterns and sculpturing complex 3D microstructures on the PEG basis. The swelling behavior of such morphological patterns was studied in detail, and it was shown that, in contrast to the pristine material, irradiated areas of the PEG films reveal an almost complete absence of the hydrogel-typical swelling behavior. The associated sealing of the irradiated areas allows a controlled deposition of objects dissolved in water, such as metal nanoparticles or fluorophores, into the surrounding, pristine areas, resulting in the formation of nanocomposite patterns. In contrast, due to the distinct protein-repelling properties of the PEG films, proteins are exclusively adsorbed onto the irradiated areas. This makes such films a suitable platform to prepare protein-affinity patterns in a protein-repelling background.

  2. Simple synthesis of smart magnetically driven fibrous films for remote controllable oil removal

    NASA Astrophysics Data System (ADS)

    Wu, Jing; Wang, Nü; Zhao, Yong; Jiang, Lei

    2015-01-01

    Inspired by the marine mussel adhesive protein, smart, magnetically controllable, oil adsorption nanofibrous materials were successfully fabricated in this research. Taking advantage of the properties of dopamine whose molecular structure mimics the single unit of the marine mussel adhesive protein and can be polymerized in alkaline solution forming a ``glue'' layer on many kinds of material surfaces, magnetic iron(ii, iii) oxide (Fe3O4) nanoparticles were easily and robustly anchored on to electrospun poly(vinylidene fluoride) fibrous films. After fluorination, the as-prepared hierarchical structured films exhibited superhydrophobicity, superoleophilicity and an excellent oil adsorption capacity from water. Importantly, because of the magnetically controllable property endowed by the Fe3O4 nanoparticles, such fibrous films act as a ``smart magnetically controlled oil removal carrier'', which effectively overcome the drawbacks of other in situ oil adsorbant materials and can also be easily recovered. This work provides a simple strategy to fabricate magnetic responsive intelligent oil removal materials, which will find broad applications in complex environment oil-water separation.Inspired by the marine mussel adhesive protein, smart, magnetically controllable, oil adsorption nanofibrous materials were successfully fabricated in this research. Taking advantage of the properties of dopamine whose molecular structure mimics the single unit of the marine mussel adhesive protein and can be polymerized in alkaline solution forming a ``glue'' layer on many kinds of material surfaces, magnetic iron(ii, iii) oxide (Fe3O4) nanoparticles were easily and robustly anchored on to electrospun poly(vinylidene fluoride) fibrous films. After fluorination, the as-prepared hierarchical structured films exhibited superhydrophobicity, superoleophilicity and an excellent oil adsorption capacity from water. Importantly, because of the magnetically controllable property endowed by the Fe3

  3. Fabrication of Self-Cleaning, Reusable Titania Templates for Nanometer and Micrometer Scale Protein Patterning.

    PubMed

    Moxey, Mark; Johnson, Alexander; El-Zubir, Osama; Cartron, Michael; Dinachali, Saman Safari; Hunter, C Neil; Saifullah, Mohammad S M; Chong, Karen S L; Leggett, Graham J

    2015-06-23

    The photocatalytic self-cleaning characteristics of titania facilitate the fabrication of reuseable templates for protein nanopatterning. Titania nanostructures were fabricated over square centimeter areas by interferometric lithography (IL) and nanoimprint lithography (NIL). With the use of a Lloyd's mirror two-beam interferometer, self-assembled monolayers of alkylphosphonates adsorbed on the native oxide of a Ti film were patterned by photocatalytic nanolithography. In regions exposed to a maximum in the interferogram, the monolayer was removed by photocatalytic oxidation. In regions exposed to an intensity minimum, the monolayer remained intact. After exposure, the sample was etched in piranha solution to yield Ti nanostructures with widths as small as 30 nm. NIL was performed by using a silicon stamp to imprint a spin-cast film of titanium dioxide resin; after calcination and reactive ion etching, TiO2 nanopillars were formed. For both fabrication techniques, subsequent adsorption of an oligo(ethylene glycol) functionalized trichlorosilane yielded an entirely passive, protein-resistant surface. Near-UV exposure caused removal of this protein-resistant film from the titania regions by photocatalytic degradation, leaving the passivating silane film intact on the silicon dioxide regions. Proteins labeled with fluorescent dyes were adsorbed to the titanium dioxide regions, yielding nanopatterns with bright fluorescence. Subsequent near-UV irradiation of the samples removed the protein from the titanium dioxide nanostructures by photocatalytic degradation facilitating the adsorption of a different protein. The process was repeated multiple times. These simple methods appear to yield durable, reuseable samples that may be of value to laboratories that require nanostructured biological interfaces but do not have access to the infrastructure required for nanofabrication.

  4. Structure and Dynamics of Proteins Adsorbed to Biomaterial Interfaces

    SciTech Connect

    Drobny, Gary P.; Long, Joanna R.; Shaw, Wendy J.; Cotten, Myriam L.; Stayton, Partick S.

    2002-10-31

    Biomineralization, defined as the organized deposition of inorganic materials in the cellular or extracellular matrix, may be as simple a process as the formation of an iron oxide crystal in the vesicle of a magnetobacterium, or as complex a process as the formation of the intricate calcium carbonate and calcium phosphate structures found in marine coccoliths, invertebrate shells, vertebrate skeletons and teeth. The phenomenon of Biomineralization has attracted a great deal of attention recently from the materials science community, which seeks to understand the way in which inorganic biological composites are synthesized and processed in nature.

  5. Negatively charged ions on Mg(0001) surfaces: appearance and origin of attractive adsorbate-adsorbate interactions.

    PubMed

    Cheng, Su-Ting; Todorova, Mira; Freysoldt, Christoph; Neugebauer, Jörg

    2014-09-26

    Adsorption of electronegative elements on a metal surface usually leads to an increase in the work function and decrease in the binding energy as the adsorbate coverage rises. Using density-functional theory calculations, we show that Cl adsorbed on a Mg(0001) surface complies with these expectations, but adsorption of {N,O,F} causes a decrease in the work function and an increase in the binding energy. Analyzing the electronic structure, we show that the presence of a highly polarizable electron spill-out in front of Mg(0001) causes this unusual adsorption behavior and is responsible for the appearance of a hitherto unknown net-attractive lateral electrostatic interaction between same charged adsorbates.

  6. Conformation transition in silk protein films monitored by time-resolved Fourier transform infrared spectroscopy: effect of potassium ions on Nephila spidroin films.

    PubMed

    Chen, Xin; Knight, David P; Shao, Zhengzhong; Vollrath, Fritz