Sample records for anti-treponema pallidum antibodies

  1. Immunoglobulin G subclasses of fluorescent anti-Treponema pallidum antibodies: evidence for sequential development of specific anti-T. pallidum immunoglobulin G responses in patients with early syphilis.

    PubMed Central

    van der Sluis, J J; van Reede, E C; Boer, M

    1986-01-01

    The development of immunoglobulin G (IgG) subclass-specific anti-Treponema pallidum antibodies during the course of syphilis in humans was studied with sera from 50 untreated male patients. The patients were divided into five diagnosis groups. In the fluorescent treponemal antibody test, which delineates the presence of cross-reacting antibodies, as well as specific antitreponema antibodies, IgG1, IgG2, and IgG3 subclass antibodies were already present during the seronegative primary stage. Specific antibodies, which were detected by the fluorescent treponemal antibody absorption test, were first present during the serotype-variable primary stage. These antibodies were almost exclusively of the IgG1 and IgG3 subclasses. In later stages, antibodies of other subclasses were detectable. Titration of IgG1 antitreponema antibodies in three electrophoretically different IgG fractions revealed an asymmetric distribution in these fractions during primary syphilis. The antibodies were largely confined to the most basic fraction during primary syphilis. A sudden change in the distribution was noted between the end of the primary stage and the secondary stage; an even distribution of IgG1 antitreponema antibodies existed in the late latent stage. These findings confirm and extend previous results from our laboratory. The development of antibodies detected by both tests is discussed in terms of a sequential stimulation of the immune system due to the presence of an extracellular layer covering the treponemas or, alternatively, in terms of a suppression of the immune response during early syphilis. PMID:3531229

  2. Evaluation of the HISCL Anti-Treponema pallidum Assay as a Screening Test for Syphilis.

    PubMed

    An, Jingna; Chen, Qixia; Liu, Qianqian; Rao, Chenli; Li, Dongdong; Wang, Tingting; Tao, Chuanmin; Wang, Lanlan

    2015-07-01

    The resurgence of syphilis in recent years has become a serious threat to public health worldwide, and the serological detection of specific antibodies against Treponema pallidum remains the most reliable method for laboratory diagnosis of syphilis. This study examined the performance of the recently launched HISCL anti-Treponema pallidum (anti-TP) assay as a screening test for syphilis in a high-volume laboratory. The HISCL anti-TP assay was tested in 300 preselected syphilis-positive samples, 704 fresh syphilis-negative samples, 48 preselected potentially interfering samples, and 30 "borderline" samples and was compared head to head with the commercially available Lumipulse G TP-N. In this study, the HISCL anti-TP assay was in perfect agreement with the applied testing algorithms with an overall agreement of 100%, comparable to that of Lumipulse G TP-N (99.63%). The sensitivity and specificity of the HISCL anti-TP assay were 100% (95% confidence interval [CI], 98.42% to 100%) and 100% (95% CI, 99.37% to 100%), respectively. Considering the excellent ease of use and automation, high throughput, and its favorable sensitivity and specificity, the HISCL anti-TP assay may represent a new choice for syphilis screening in high-volume laboratories. PMID:25972403

  3. Validation of Serological Testing for Anti-Treponema pallidum from Postmortem Blood on the Siemens-BEP®-III Automatic System

    PubMed Central

    Kalus, Ulrich; Wilkemeyer, Ina; Pruss, Axel; Caspari, Gregor

    2013-01-01

    Summary Background Infectious disease marker testing is obligatory for the release of human tissue for transplantation. Most CE-marked tests are not validated for postmortem blood. In a previous study we have validated the testing for anti-HIV-1/2, anti-HCV, HBsAg, and anti-HBc. Here, we present the validation of testing for antibodies against T. pallidum, which is the last marker obligatory for tissue release for transplantation. Methods 17 samples of postmortem sera and 10 samples of both pre-und postmortem sera were obtained from cornea donors and tested for anti-T. pallidum on the Siemens-BEP-III-System. These sera were spiked with anti-T. pallidum-positive standard sera in concentrations which give low- and high-positive results at the respective dilution. Results Two of the unspiked postmortem sera were false-positive most likely due to intense hemolysis (free hemoglobin > 50 mg/dl). Of the 25 negative postmortem sera, none of the spiked samples was false-negative after 0, 24 and 60 h. Conclusion There is no indication that postmortem samples give false-negative or false-positive results with the test system and test kits used in cases of low hemolysis. The procedure described might serve as a model for validating other test kits on postmortem samples. PMID:24474889

  4. Quantitative microhaemagglutination assay for Treponema pallidum antibodies in experimental syphilis.

    PubMed Central

    Tight, R R; White, A C

    1980-01-01

    The quantitative microhaemagglutination assay for Treponema pallidum antibodies (MHA-TP) was studies in 52 untreated and treated rabbits with experimental syphilis. Rabbits with incubating experimental syphilis were cured or inadequately treated with penicillin G and some cured rabbits were later reinfected. MHA-TP conversion occurred within 45 days in untreated rabbits. Titres reached peak levels about four months after inoculation and remained relatively high for up to two years. The quantitative MHA-TP test differentiated between rabbits cured of experimental incubating syphilis and those untreated and inadequately treated. MHA-TP titres decreased after treatment given six or 12 months after inoculation but reversion did not occur. MHA-TP conversion or significant increases in titre occurred as soon as seven days after reinfection and preceded corresponding changes in a quantitative non-treponemal test. The MHA-TP is useful as a screening test for treponemal antibodies in rabbits. The quantitative MHA-TP in humans after treatment for syphilis and reinfection deserves further study. PMID:7000307

  5. Microcapsule agglutination test for Treponema pallidum antibodies. A new serodiagnostic test for syphilis.

    PubMed Central

    Kobayashi, S; Yamaya, S I; Sugahara, T; Matuhasi, T

    1983-01-01

    For the serodiagnosis of syphilis a quantitative passive agglutination (MCA-TP) test for antibodies to Treponema pallidum was performed with chemically stable microcapsules with no antigenic activity instead of with conventional sheep erythrocytes. The microcapsules were easily sensitised with the antigen of sonicated Treponema pallidum by treatment with glutaraldehyde. Compared with the Treponema pallidum haemagglutination test (TPHA) the MCA-TP test was superior for detecting cases of primary syphilis. Furthermore, the decrease in antibody titre during treatment was more evident in this test than in the FTA-ABS or the TPHA tests. The MCA-TP test performed on IgM and IgG gel-filtered fractions of sera from patients with syphilis proved that the sensitised microcapsule antigen reacted sharply with the IgM antibodies specific to syphilis. PMID:6337678

  6. Detection of immunoglobulin M antibodies to Treponema pallidum in a modified enzyme-linked immunosorbent assay

    Microsoft Academic Search

    F. Mtiller; M. Moskophidis; H.-L. Borkhardt

    1987-01-01

    The indirect enzyme-linked immunosorbent assay (ELISA) for detection of immunoglobulin M (IgM) antibodies toTreponema pallidum in sera of syphilitic patients is complicated by false positive reactions due to the interference of IgM rheumatoid factor (IgM-RF) activity and the presence of treponemal IgG antibodies. Another source of error producing false negative results is the competition between treponemal IgG and IgM antibodies

  7. Laboratory Evaluation of Three Rapid Diagnostic Tests for Dual Detection of HIV and Treponema pallidum Antibodies

    PubMed Central

    Woo, Jennifer S.; Chung, Jun Ho; Sokovic, Anita; Bristow, Claire C.; Klausner, Jeffrey D.

    2014-01-01

    The performance of three research-use-only, dual HIV and syphilis rapid diagnostic tests (RDTs) was evaluated for 150 patient serum samples and compared to reference HIV and Treponema pallidum antibody detection methods. The RDTs performed comparably, with sensitivities of 93 to 99% and specificities of 97 to 100%. The kappa statistic between the RDTs was 0.95. PMID:25297332

  8. Validation of Serological Tests for the Detection of Antibodies Against Treponema pallidum in Nonhuman Primates

    PubMed Central

    Knauf, Sascha; Dahlmann, Franziska; Batamuzi, Emmanuel K.; Frischmann, Sieghard; Liu, Hsi

    2015-01-01

    There is evidence to suggest that the yaws bacterium (Treponema pallidum ssp. pertenue) may exist in non-human primate populations residing in regions where yaws is endemic in humans. Especially in light of the fact that the World Health Organizaiton (WHO) recently launched its second yaws eradication campaign, there is a considerable need for reliable tools to identify treponemal infection in our closest relatives, African monkeys and great apes. It was hypothesized that commercially available serological tests detect simian anti-T. pallidum antibody in serum samples of baboons, with comparable sensitivity and specificity to their results on human sera. Test performances of five different treponemal tests (TTs) and two non-treponemal tests (NTTs) were evaluated using serum samples of 57 naturally T. pallidum-infected olive baboons (Papio anubis) from Lake Manyara National Park in Tanzania. The T. pallidum particle agglutination assay (TP-PA) was used as a gold standard for comparison. In addition, the overall infection status of the animals was used to further validate test performances. For most accurate results, only samples that originated from baboons of known infection status, as verified in a previous study by clinical inspection, PCR and immunohistochemistry, were included. All tests, TTs and NTTs, used in this study were able to reliably detect antibodies against T. pallidum in serum samples of infected baboons. The sensitivity of TTs ranged from 97.7-100%, while specificity was between 88.0-100.0%. The two NTTs detected anti-lipoidal antibodies in serum samples of infected baboons with a sensitivity of 83.3% whereas specificity was 100%. For screening purposes, the TT Espline TP provided the highest sensitivity and specificity and at the same time provided the most suitable format for use in the field. The enzyme immune assay Mastblot TP (IgG), however, could be considered as a confirmatory test. PMID:25803295

  9. Monoclonal antibody selection and analysis of a recombinant DNA-derived surface immunogen of Treponema pallidum expressed in Escherichia coli.

    PubMed Central

    Swancutt, M A; Twehous, D A; Norgard, M V

    1986-01-01

    Monoclonal antibodies directed against a 34-kilodalton (kDa) surface immunogen of Treponema pallidum were used to select 12 unique T. pallidum DNA-containing Escherichia coli recombinant clones expressing the recombinant form of the 34-kDa immunogen. The phenotype of the clones was dependent on the presence of recombinant plasmids in the host cell. Restriction enzyme analyses and Southern hybridization of plasmid DNA demonstrated that all recombinant clones contained common DNA sequences of T. pallidum origin. Further hybridization analyses revealed that the cloned T. pallidum DNA sequences were an accurate representation of the T. pallidum genomic DNA arrangement. Purified immunoglobulin G (IgG) from pooled immune rabbit serum reacted with the clones, while IgG from pooled normal rabbit serum did not. Results of immunological experiments and Southern hybridization indicated that a similar 34-kDa immunogen was present in T. pallidum subsp. pertenue, but it was absent from four species of nonpathogenic treponemes tested, as well as from homogenates of normal rabbit testicular tissue. Metabolic labeling of the E. coli clones with [35S]methionine followed by radioimmunoprecipitation with monoclonal antibodies revealed that the 35S-labeled recombinant and 125I-labeled native (T. pallidum) forms of the antigen had identical electrophoretic mobilities. The production of a complete antigen by E. coli was independent of the orientation of the foreign gene sequence with respect to vector DNA. T. pallidum also produced an apparently identical immunoprecipitable 34-kDa antigen after metabolic labeling with [35S]methionine in the presence of cycloheximide. The apparent specificity of the 34-kDa immunogen for pathogenic treponemes and its native cell surface association on T. pallidum justifies a more intense study of this antigen and its corresponding gene. Images PMID:3514451

  10. Evaluation of the Determine Syphilis TP assay for the detection of antibodies against Treponema pallidum for the serodiagnosis of syphilis.

    PubMed

    Zhuang, Y-H; Tian, Y; Chen, Y; Tang, J; Wang, J-Q; Li, P; Li, Q; Jiang, Y-Q

    2012-06-01

    Currently, infectious syphilis has been resurgent in China and has become a significant public health problem. The rapid expansion of syphilis screening programs is urgently required. In the present study, the performance of the Determine Syphilis TP assay (Determine TP assay) for the detection of antibodies against Treponema pallidum (T. pallidum) for syphilis serodiagnosis was evaluated. In total, 300 serum samples were tested for the presence of treponemal-specific antibodies using the Treponema pallidum particle agglutination (TPPA) assay, the Determine TP assay, and the InTec immunochromatography assay (InTec assay). The Determine TP assay detected 99, 11, and 5 positive results, whereas the InTec assay detected 97, 3, and 3 positive samples from group I (100 TPPA-positive sera), group II (13 TPPA 1:80 +/- sera), and group III (187 TPPA-negative sera), respectively. The sensitivity, specificity, and the rate concordant with TPPA for the Determine TP assay were 97.35, 98.91, and 97.33%, respectively. In comparison to the TPPA, the Determine TP assay is simple to perform and time-saving, making it a favorable alternative for the detection of T. pallidum-specific antibodies where other T. pallidum-specific confirmatory tests are not available. In addition, this rapid treponemal test promotes prompt treatment for syphilis by providing early laboratory diagnosis. PMID:21866323

  11. Prevalence of antibodies against Treponema pallidum among HIV-positive patients in a tertiary care hospital in Mexico.

    PubMed

    Mata-Marín, José Antonio; Sandoval-Sánchez, Juan Joel; Huerta-García, Gloria; Arroyo-Anduiza, Carla Ileana; Alcalá-Martínez, Enrique; Mata-Marín, Luis Alberto; Sandoval-Ramirez, Jorge Luis; Gaytán-Martínez, Jesús

    2015-02-01

    Our objective was to determine the seroprevalence of syphilis among HIV-infected patients in a tertiary care hospital in Mexico City. A cross-sectional study was developed, and 318 HIV-positive patients were evaluated from January to February 2013 at Hospital de Infectología, National Medical Center 'La Raza' (a tertiary care hospital specialising in infectious diseases in Mexico City). Laboratory data were screened for the detection of antibodies against Treponema pallidum. Patients completed a questionnaire relating to socio-demographic data and factors associated with syphilis. Of the 318 patients, 83% were men. The mean age ± SD was 36?±?11 years; 52% were men who have sex with men and 47% had undertaken higher education. The overall seroprevalence of syphilis among these patients was 25% (95% confidence interval 21%, 30%). Men who have sex with men had a significantly higher seroprevalence (30% vs. 15%, p?=?0.009). We conclude that, in Mexico, there is a high seroprevalence of syphilis antibodies in HIV-infected patients and that men who have sex with men are the group most affected. PMID:24713227

  12. Further evaluation of the characteristics of Treponema pallidum-specific IgM antibody in syphilis serofast reaction patients.

    PubMed

    Lin, Li-Rong; Zheng, Wei-Hong; Tong, Man-Li; Fu, Zuo-Gen; Liu, Gui-Li; Fu, Jian-Guo; Zhang, Dai-Wei; Yang, Tian-Ci; Liu, Li-Li

    2011-11-01

    Syphilis serofast reaction (SSR) is common in clinical work. From June 2005 to May 2009, 1208 syphilis patients were chosen for research by the Xiamen Center of Clinical Laboratory in China. Serologic tests were performed with toluidine red unheated serum test (TRUST) and Treponema pallidum particle agglutination (TPPA). Then, T. pallidum-specific IgM antibody (TP-IgM) was detected with fluorescent treponemal antibody absorption (FTA-Abs) and TPPA. In this study, patients were divided into the following experimental groups according to the results of TRUST and TPPA: (1) the SSR group consisted of 411 cases with (+) TRUST and (+) TPPA, and without clinical manifestations after 1 year of recommended syphilis treatment; (2) the serum cure group, which was further subdivided into group A consisting of 251cases with (-) TRUST and (+) TPPA; (3) group B consisting of 546 cases with (-) TRUST and (-) TPPA; and (4) the blood donor control group which consisted of 100 cases. We demonstrated that a total of 136 cases (33.09%) of 411 SSR patients were TP-IgM positive by TPPA, and this percentage was markedly higher than that in serum cure group A (9.16%). FTA-Abs analyses revealed similar results. All samples in serum cure group B and the control group were TP-IgM negative, which is identical to our previous report. The present study also indicated that the TP-IgM positive rate was not significantly different among patients with different ages, genders, and clinical phases after 1 year of recommended therapy. From the total of 1208 syphilis patients, 289 were randomly selected for TP-DNA detection by fluorescence quantitative polymerase chain reaction, and the positive rate of TP-DNA was 32.53%, which was slightly higher than that of FTA-Abs TP-IgM, and no statistically significant difference by chi-square tests, indicating the TP-DNA result is preferably consistent with FTA-Abs and supporting our deduction that TP-IgM could be used as a serologic marker for the relapse and infection of syphilis. PMID:21899981

  13. Monoclonal antibodies to the recombinant protein TmpA of the Treponema pallidum.

    PubMed

    Brito Moreno, Adys I; Acosta Bas, Carmen; Rodríguez, Maya; Baluja Conde, Ileana B; Feal Carballo, Sadys; Martínez, Luisa

    2003-12-01

    Spleen cells from BALB/c mice immunized with recombinant TmpA were fused with mouse myeloma cells (P3/X63-Ag8), and five hybridomas secreting monoclonal antibodies were obtained. These hybridomas specifically recognize TmpA and do not cross-react with other molecules such as recombinant HBsAg of HBV and synthetic HCV core peptides. The monoclonal antibodies were IgG1 subclass and ascitic fluid from these hybridomas was purified by affinity chromatography on Protein A-Sepharose CL-4B column to isolate the IgG1 active fraction. The affinity constant of these monoclonal antibodies ranged from 6.4 x 10(8) and 1.73 x 10(10) M(-1). PMID:14683600

  14. Characterization of Treponema pallidum particle agglutination assay-negative sera following screening by treponemal total antibody enzyme immunoassays.

    PubMed

    Maple, P A C; Ratcliffe, D; Smit, E

    2010-11-01

    Following a laboratory audit, a significant number of Treponema pallidum particle agglutination assay (TPPA)-negative sera were identified when TPPA was used as a confirmatory assay of syphilis enzyme immunoassay (EIA) screening-reactive sera (SSRS). Sera giving such discrepant results were further characterized to assess their significance. A panel of 226 sera was tested by the Abbott Murex ICE Syphilis EIA and then by the Newmarket Syphilis EIA II. TPPA testing was performed on 223 sera. Further testing by the Venereal Disease Research Laboratory (VDRL) test, the Mercia Syphilis IgM EIA, the fluorescent treponemal antibody (FTA-ABS) assay, and INNO-LIA immunoblotting was undertaken in discrepant cases. One hundred eighty-seven of 223 (83.8%) SSRS were TPPA reactive, while 26 (11.6%) sera which were reactive in both the ICE and Newmarket EIAs were nonreactive by TPPA. The majority (68%) of the TPPA-discrepant sera were from HIV-positive patients and did not represent early acute cases, based on previous or follow-up samples, which were available for 22/26 samples. FTA-ABS testing was performed on 24 of these sera; 14 (58.3%) were FTA-ABS positive, and 10 (41.7%) were FTA-ABS negative. Twenty-one of these 26 sera were tested by INNO-LIA, and an additional 4 FTA-ABS-negative samples were positive. In this study, significant numbers (18/26) of SSRS- and TPPA-negative sera were shown by further FTA-ABS and LIA (line immunoblot assay) testing to be positive. The reason why certain sera are negative by TPPA but reactive by treponemal EIA and other syphilis confirmatory assays is not clear, and these initial findings should be further explored. PMID:20844087

  15. Effect of passive immunization with purified specific or cross-reacting immunoglobulin G antibodies against Treponema pallidum on the course of infection in guinea pigs.

    PubMed Central

    Wicher, K; Zabek, J; Wicher, V

    1992-01-01

    Whole immune serum or highly purified immunoglobulin G (IgG) antibodies to Treponema pallidum exhaustively adsorbed with three strains of nonpathogenic treponemes (TPI-IgG) were used for passive immunization of inbred strain 2 guinea pigs before and after intradermal challenge with 3.4 x 10(7) virulent T. pallidum Nichols organisms. Before challenge, control animals received a similarly purified IgG fraction containing either a cocktail of antibodies against three nonpathogenic treponemes (NPTI-IgG) or IgG prepared from normal guinea pig serum (NGPS-IgG). The purified fractions contained both IgG1 and IgG2 isotypes. The antibody levels (detected by fluorescent treponemal antibody test and enzyme-linked immunosorbent assay) and molecular specificities (immunoblot) of sera obtained from recipient animals before infection reflected those of the purified fractions used for immunization. Three protocols of passive immunization were used. Whole immune serum containing specific and cross-reacting antibodies afforded better protection than TPI-IgG even though asymptomatic animals were not fully protected. A single intradermal injection (0.1 ml) of TPI-IgG or NPTI-IgG into one hind leg 22 h before infection at the same site provided relatively higher protection than multiple intravenous injections (total, 15 ml) of the respective individual preparations. Since purified NGPS-IgG injected in the same animals, into the opposite hind leg, failed to protect against the challenging infection, it is reasonable to assume that specific and cross-reacting antitreponemal antibodies of the IgG1 subclass, which in guinea pigs are homocytotropic, play a relevant role in local protection. Images PMID:1639492

  16. Genome Scale Identification of Treponema pallidum Antigens

    Microsoft Academic Search

    Matthew McKevitt; Mary Beth Brinkman; Melanie McLoughlin; Carla Perez; Jerrilyn K. Howell; George M. Weinstock; Steven J. Norris; Timothy Palzkill

    2005-01-01

    Antibody responses for 882 of the 1,039 proteins in the proteome of Treponema pallidum were examined. Sera collected from infected rabbits were used to systematically identify 106 antigenic proteins, including 22 previously identified antigens and 84 novel antigens. Additionally, sera collected from rabbits throughout the course of infection demonstrated a progression in the breadth and intensity of humoral immunoreactivity against

  17. Molecular Subtyping of Treponema pallidum subsp. pallidum in Lisbon, Portugal

    Microsoft Academic Search

    R. Castro; E. Prieto; M. J. Aguas; M. J. Manata; J. Botas; F. Martins Pereira

    2009-01-01

    The objectives of this study were to evaluate the reproducibility of a molecular method for the subtyping of Treponema pallidum subsp. pallidum and to discriminate strains of this microorganism from strains from patients with syphilis. We studied 212 specimens from a total of 82 patients with different stages of syphilis (14 primary, 7 secondary and 61 latent syphilis). The specimens

  18. Usefulness in clinical practice of a point-of-care rapid test for simultaneous detection of nontreponemal and Treponema pallidum-specific antibodies in patients suffering from documented syphilis.

    PubMed

    Guinard, Jérôme; Prazuck, Thierry; Péré, Hélčne; Poirier, Claire; LeGoff, Jérôme; Boedec, Erwan; Guigon, Aurélie; Day, Nesrine; Bélec, Laurent

    2013-12-01

    The usefulness of a point-of-care immunochromatographic dual test for the simultaneous detection of both nontreponemal and Treponema pallidum-specific antibodies (Chembio Diagnostics Systems Inc., Medford, NY, USA) was assessed in various situations related to syphilis, by reference to conventional syphilis serology. Thawed sera were obtained from 100 adults including 36 primary syphilis, 6 secondary syphilis, 6 re-infection, 9 recently-treated syphilis, and 43 old syphilis. Doubtful reactivities for the treponemal line were considered positive; doubtful reactivities for the nontreponemal line were considered positive only when the treponemal line was present. The sensitivity, the specificity, and its concordance to gold standard serology of treponemal line were high, around 90%. The sensitivity of nontreponemal line was 96.3%, its specificity 76.7%, and its concordance 83.4%. In conclusion, the dual rapid test from Chembio Diagnostics Systems Inc. is useful for rapid point-of-care diagnosis in the various situations encountered with patients suffering from syphilis. PMID:23999937

  19. Evaluation of a colloidal gold immunochromatography assay in the detection of Treponema pallidum specific IgM antibody in syphilis serofast reaction patients: a serologic marker for the relapse and infection of syphilis.

    PubMed

    Lin, Li-Rong; Tong, Man-Li; Fu, Zuo-Gen; Dan, Bing; Zheng, Wei-Hong; Zhang, Chang-Gong; Yang, Tian-Ci; Zhang, Zhong-Ying

    2011-05-01

    Syphilis remains as a worldwide public health problem; hence, it is necessary to develop a new diagnostic approach that is easier and faster than conventional tests. A new testing method to detect Treponema pallidum IgM (TP-IgM), named colloidal gold immunochromatography assay (GICA), is presented in place of fluorescent treponemal antibody absorption (FTA-Abs). TP-IgM was detected using GICA developed on syphilis-specific recombinant proteins TPN17 and TPN47. The FTA-Abs IgM test was set as the gold standard. A GICA TP-IgM test was performed to detect syphilis in 1208 patients who received recommended therapy for syphilis for more than 1 year at the Xiamen Center of Clinical Laboratory in China from June 2005 to May 2009. One hundred blood donors were set up as control. The sensitivity, specificity, positive predictive value, negative predictive value, positive likelihood ratio, and negative likelihood ratio were 98.21%, 99.04%, 93.75%, 99.73%, 102.3, and 0.018, respectively. Detection on 500 interference specimens indicated that the biological false-positive rate of the GICA test was extremely low and was free from other biological and chemical factors. The patients were divided into the following experimental groups based on the results of toluidine red unheated serum test (TRUST) and treponemal pallidum particle agglutination (TPPA): (1) the syphilis serofast reaction (SSR) group consisted of 411 cases with (+) TRUST and (+) TPPA, which exhibited no clinical manifestations of syphilis after 1 year of recommended syphilis treatment; (2) the serum cure group, which was further subdivided into group A, a group that consisted of 251 cases with (-) TRUST and (+) TPPA, and (3) group B, a group that consisted of 546 cases with (-) TRUST and (-) TPPA; and (4) the blood donor control group, which consisted of 100 healthy persons with (-) ELISA-TP and (-) TPPA. We used the FTA-Abs method and the GICA method to detect TP-IgM; the positive rate of TP-IgM in 411 SSR patients was 34.55% and 36.01%, respectively. However, in serum cure group A, the positive rate of TP-IgM was 10.36% and 11.16%, respectively. The ?(2) test revealed that there is a significant difference in the positive rate between these 2 groups (P < 0.01). The TP-IgM positive rate in the same group, as detected by the GICA method and the FTA-Abs method, had no significant difference in statistics. However, as detected by the GICA method and the FTA-Abs method, all the samples in serum cure group B and the control group were negative for TP-IgM. The TP-IgM-positive result demonstrated that active T. pallidum remained in the bodies of SSR patients. In summary, the characteristics of GICA TP-IgM correspond to that of FTA-Abs TP-IgM; this can be used as a serologic marker for the relapse and infection of syphilis in place of the conventional FTA-Abs IgM test. PMID:21388769

  20. Footprint of Positive Selection in Treponema pallidum subsp. pallidum Genome Sequences Suggests Adaptive Microevolution of the Syphilis Pathogen

    PubMed Central

    Centurion-Lara, Arturo; Jeffrey, Brendan M.; Le, Hoavan T.; Molini, Barbara J.; Lukehart, Sheila A.; Sokurenko, Evgeni V.; Rockey, Daniel D.

    2012-01-01

    In the rabbit model of syphilis, infection phenotypes associated with the Nichols and Chicago strains of Treponema pallidum (T. pallidum), though similar, are not identical. Between these strains, significant differences are found in expression of, and antibody responses to some candidate virulence factors, suggesting the existence of functional genetic differences between isolates. The Chicago strain genome was therefore sequenced and compared to the Nichols genome, available since 1998. Initial comparative analysis suggested the presence of 44 single nucleotide polymorphisms (SNPs), 103 small (?3 nucleotides) indels, and 1 large (1204 bp) insertion in the Chicago genome with respect to the Nichols genome. To confirm the above findings, Sanger sequencing was performed on most loci carrying differences using DNA from Chicago and the Nichols strain used in the original T. pallidum genome project. A majority of the previously identified differences were found to be due to errors in the published Nichols genome, while the accuracy of the Chicago genome was confirmed. However, 20 SNPs were confirmed between the two genomes, and 16 (80.0%) were found in coding regions, with all being of non-synonymous nature, strongly indicating action of positive selection. Sequencing of 16 genomic loci harboring SNPs in 12 additional T. pallidum strains, (SS14, Bal 3, Bal 7, Bal 9, Sea 81-3, Sea 81-8, Sea 86-1, Sea 87-1, Mexico A, UW231B, UW236B, and UW249C), was used to identify “Chicago-“ or “Nichols -specific” differences. All but one of the 16 SNPs were “Nichols-specific”, with Chicago having identical sequences at these positions to almost all of the additional strains examined. These mutations could reflect differential adaptation of the Nichols strain to the rabbit host or pathoadaptive mutations acquired during human infection. Our findings indicate that SNPs among T. pallidum strains emerge under positive selection and, therefore, are likely to be functional in nature. PMID:22720110

  1. Polypeptides of Treponema pallidum: progress toward understanding their structural, functional, and immunologic roles. Treponema Pallidum Polypeptide Research Group.

    PubMed Central

    Norris, S J

    1993-01-01

    Treponema pallidum subsp. pallidum, the spirochete that causes syphilis, is unusual in a number of respects, including its small genome size, inability to grow under standard in vitro culture conditions, microaerophilism, apparent paucity of outer membrane proteins, structurally complex periplasmic flagella, and ability to evade the host immune responses and cause disease over a period of years to decades. Many of these attributes are related ultimately to its protein content. Our knowledge of the activities, structure, and immunogenicity of its proteins has been expanded by the application of recombinant DNA, hybridoma, and structural fractionation techniques. The purpose of this monograph is to summarize and correlate this new information by using two-dimensional gel electrophoresis, monoclonal antibody reactivity, sequence data, and other properties as the bases of polypeptide identification. The protein profiles of the T. pallidum subspecies causing syphilis, yaws, and endemic syphilis are virtually indistinguishable but differ considerably from those of other treponemal species. Among the most abundant polypeptides are a group of lipoproteins of unknown function that appear to be important in the immune response during syphilitic infection. The periplasmic flagella of T. pallidum and other spirochetes are unique with regard to their protein content and ultrastructure, as well as their periplasmic location. They are composed of three core proteins (homologous to the other members of the eubacterial flagellin family) and a single, unrelated sheath protein; the functional significance of this arrangement is not understood at present. Although the bacterium contains the chaperonins GroEL and DnaK, these proteins are not under the control of the heat shock regulon as they are in most organisms. Studies of the immunogenicity of T. pallidum proteins indicate that many may be useful for immunodiagnosis and immunoprotection. Future goals in T. pallidum polypeptide research include continued elucidation of their structural locations and functional activities, identification and characterization of the low-abundance outer membrane proteins, further study of the immunoprotective and immunodiagnostic potential of T. pallidum proteins, and clarification of the roles of treponemal proteins in pathogenesis. Images PMID:8246847

  2. Specific immunofluorescent staining of pathogenic treponemes with a monoclonal antibody.

    PubMed Central

    Ito, F; Hunter, E F; George, R W; Pope, V; Larsen, S A

    1992-01-01

    Two hybrid cell lines which produced mouse monoclonal antibody to the DAL-1 street strain of Treponema pallidum subsp. pallidum were established. These monoclonal antibodies strongly reacted with T. pallidum subsp. pallidum (Nichols strain, DAL-1, and two other street strains, strains MN-1 and MN-3) and T. pallidum subsp. pertenue by indirect microimmunofluorescent antibody and enzyme-linked immunosorbent assay techniques, but they did not react with normal rabbit testicular tissue. These monoclonal antibodies did not react with nonpathogenic treponemes, such as T. phagedenis Reiter, T. denticola MRB, T. refringens Noguchi, or other spirochetes, such as Borrelia burgdorferi and Leptospira interrogans serovar pomona in microimmunofluorescent antibody smear slides or in Western blots (immunoblots). While unlabeled antibodies are useful for investigating the antigenic structures of T. pallidum, we labeled these monoclonal antibodies with fluorescein isothiocyanate and used them for diagnosing syphilis by direct staining of lesion exudate or T. pallidum subsp. pallidum in formalin-fixed tissues from patients suspected of having syphilis. Both monoclonal antibodies were directed against antigens of T. pallidum subsp. pallidum with a molecular weight of 37,000 as determined by the Western blotting technique. Images PMID:1374079

  3. Expression of Treponema pallidum antigens in Escherichia coli K-12.

    PubMed Central

    Stamm, L V; Folds, J D; Bassford, P J

    1982-01-01

    A colony bank of recombinant plasmids harboring Treponema pallidum DNA inserts has been established in Escherichia coli K-12. By using an in situ immunoassay, we identified four E. coli clones that expressed T. pallidum antigens. Thus, recombinant DNA technology may provide powerful new tools for studying the pathogenesis of T. pallidum infection. Images FIG. 1 FIG. 2 FIG. 3 PMID:7047395

  4. Characterization of the low-molecular-mass proteins of virulent Treponema pallidum.

    PubMed Central

    Stamm, L V; Parrish, E A

    1994-01-01

    We previously demonstrated that Treponema pallidum cells incubated in vitro in the presence of heat-inactivated normal rabbit serum (HINRS) synthesize, in very small quantities, several pathogen-specific, low-molecular-mass proteins that appear to be localized extracellularly. In this study, we have taken advantage of our ability to metabolically radiolabel T. pallidum cells to high specific activity to further characterize these antigens. We found that the low-molecular-mass proteins are not related to the 15- and 17-kDa detergent-phase proteins (J. D. Radolf, N. R. Chamberlain, A. Clausell, and M. V. Norgard, Infect. Immun. 56:490-498, 1988). The low-molecular-mass proteins did not incorporate 3H-labeled fatty acids and were not precipitated by rabbit immunoglobulin G (IgG) antibodies directed against glutathione S-transferase fusions to the nonlipidated 15- and 17-kDa proteins. We prepared polyclonal antisera to the low-molecular-mass proteins by immunizing two rabbits with the concentrated supernatant of T. pallidum cells. IgG antibodies present in the sera of both rabbits precipitated a 21.5-kDa protein from solubilized extracts of T. pallidum supernatant and cells. IgG antibodies in the serum of the second rabbit precipitated an additional 15.5-kDa low-molecular-mass protein only from solubilized extracts of supernatant. While investigating the effect of eliminating HINRS from the extraction medium, we observed that the low-molecular-mass proteins remained associated with treponemal cells that were incubated in the absence of HINRS. These proteins could be eluted from the cells by the addition of HINRS or rabbit serum albumin, suggesting that they are located on or near the treponemal cell surface. The 15.5- and 21.5-kDa low-molecular-mass proteins were not washed off treponemal cells with buffer containing 1 M KCl. Experiments employing selective solubilization of the T. pallidum outer membrane with 0.1% Triton X-114 and proteinase K accessibility indicated that the 15.5-kDa protein, but not the 21.5-kDa protein, is cell surface exposed. Images PMID:8262639

  5. Surface Immunolabeling and Consensus Computational Framework To Identify Candidate Rare Outer Membrane Proteins of Treponema pallidum? †

    PubMed Central

    Cox, David L.; Luthra, Amit; Dunham-Ems, Star; Desrosiers, Daniel C.; Salazar, Juan C.; Caimano, Melissa J.; Radolf, Justin D.

    2010-01-01

    Treponema pallidum reacts poorly with the antibodies present in rabbit and human syphilitic sera, a property attributed to the paucity of proteins in its outer membrane. To better understand the basis for the syphilis spirochete's “stealth pathogenicity,” we used a dual-label, 3-step amplified assay in which treponemes encapsulated in gel microdroplets were probed with syphilitic sera in parallel with anti-FlaA antibodies. A small (approximately 5 to 10%) but reproducible fraction of intact treponemes bound IgG and/or IgM antibodies. Three lines of evidence supported the notion that the surface antigens were likely ?-barrel-forming outer membrane proteins (OMPs): (i) surface labeling with anti-lipoidal (VDRL) antibodies was not observed, (ii) immunoblot analysis confirmed prior results showing that T. pallidum glycolipids are not immunoreactive, and (iii) labeling of intact organisms was not appreciably affected by proteinase K (PK) treatment. With this method, we also demonstrate that TprK (TP0897), an extensively studied candidate OMP, and TP0136, a lipoprotein recently reported to be surface exposed, are both periplasmic. Consistent with the immunolabeling studies, TprK was also found to lack amphiphilicity, a characteristic property of ?-barrel-forming proteins. Using a consensus computational framework that combined subcellular localization and ?-barrel structural prediction tools, we generated ranked groups of candidate rare OMPs, the predicted T. pallidum outer membrane proteome (OMPeome), which we postulate includes the surface-exposed molecules detected by our enhanced gel microdroplet assay. In addition to underscoring the syphilis spirochete's remarkably poor surface antigenicity, our findings help to explain the complex and shifting balance between pathogen and host defenses that characterizes syphilitic infection. PMID:20876295

  6. Reactivity of lymphocytes from patients with syphilis towards T. pallidum antigen in the leucocyte migration and lymphocyte transformation tests.

    PubMed Central

    From, E; Thestrup-Pedersen, K; Thulin, H

    1976-01-01

    The reactivity of lymphocytes to Treponema pallidum antigen was studied before and after treatment in nine patients with early syphilis using a leucocyte migration test and a lymphocyte transformation test. Lymphocyte reactivity was also investigated in six patients treated for syphilis within the last 4 years, and in five untreated patients with a positive result to the T. pallidum immobilization test, but negative results to other serum tests for syphilis antibodies and without any known exposure to risk of infection by syphilis. Ten seronegative patients with different dermatological disorders served as a control group. A significant increase in lymphocyte reactivity to T. pallidum antigen was recorded in both tests in vitro after treatment. There was no difference in lymphocyte reactivity to T. pallidum antigen between the other patients studied and the control group. In early syphilis the spontaneous migration was found to be inhibited before treatment. Tuberculin skin tests were also performed and found to be suppressed in patients with primary and secondary syphilis. No difference in phytohaemagglutinin response was found between any of the groups. Plasma from patients with primary and secondary syphilis was found to change the in vitro reactivity of normal lymphocytes when stimulated with different mitogens. PMID:786437

  7. Rapid Treponema pallidum clearance from blood and ulcer samples following single dose benzathine penicillin treatment of early syphilis.

    PubMed

    Tipple, Craig; Jones, Rachael; McClure, Myra; Taylor, Graham

    2015-02-01

    Currently, the efficacy of syphilis treatment is measured with anti-lipid antibody tests. These can take months to indicate cure and, as a result, syphilis treatment trials require long periods of follow-up. The causative organism, Treponema pallidum (T. pallidum), is detectable in the infectious lesions of early syphilis using DNA amplification. Bacteraemia can likewise be identified, typically in more active disease. We hypothesise that bacterial clearance from blood and ulcers will predict early the standard serology-measured treatment response and have developed a qPCR assay that could monitor this clearance directly in patients with infectious syphilis. Patients with early syphilis were given an intramuscular dose of benzathine penicillin. To investigate the appropriate sampling timeframe samples of blood and ulcer exudate were collected intensively for T. pallidum DNA (tpp047 gene) and RNA (16S rRNA) quantification. Sampling ended when two consecutive PCRs were negative. Four males were recruited. The mean peak level of T. pallidum DNA was 1626 copies/ml whole blood and the mean clearance half-life was 5.7 hours (std. dev. 0.53). The mean peak of 16S rRNA was 8879 copies/ml whole blood with a clearance half-life of 3.9 hours (std. dev. 0.84). From an ulcer, pre-treatment, 67,400 T. pallidum DNA copies and 7.08 x 107 16S rRNA copies were detected per absorbance strip and the clearance half-lives were 3.2 and 4.1 hours, respectively. Overall, T. pallidum nucleic acids were not detected in any sample collected more than 56 hours (range 20-56) after treatment. All patients achieved serologic cure. In patients with active early syphilis, measuring T. pallidum levels in blood and ulcer exudate may be a useful measure of treatment success in therapeutic trials. These laboratory findings need confirmation on a larger scale and in patients receiving different therapies. PMID:25700164

  8. 21 CFR 866.3830 - Treponema pallidum tre-ponemal test reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...Treponema pallidum tre-ponemal test reagents. (a) Identification...Treponema pallidum treponemal test reagents are devices that consist...antisera and all control reagents (standardized reagents with which test results are compared)...

  9. Topography and chemoarchitecture of the striatum and pallidum in a monotreme, the short-beaked echidna (Tachyglossus aculeatus).

    PubMed

    Ashwell, K W S

    2008-09-01

    The topography and chemoarchitecture of the striatum and pallidum in a monotreme, the short-beaked echidna (Tachyglossus aculeatus) have been studied using Nissl staining in conjunction with myelin staining, enzyme reactivity to acetylcholinesterase and NADPH diaphorase, and immunoreactivity to parvalbumin, calbindin, calretinin, tyrosine hydroxylase, neuropeptide Y, and neurofilament protein (SMI-32 antibody). All those components of the striatum and pallidum found in eutherian mammals could also be identified in the echidna's brain, with broad chemoarchitectural similarities to those regions in eutherian brains also apparent. There was a clear chemoarchitectural gradient visible with parvalbumin immunoreactivity of neurons and fibers, suggesting a subdivision of the echidna caudatoputamen into weakly reactive rostrodorsomedial and strongly reactive caudoventrolateral components. This may, in turn, relate to subdivision into associative versus sensorimotor CPu and reflect homology to the caudate and putamen of primates. Moreover, the chemoarchitecture of the echidna striatum suggested the presence of striosome-matrix architecture. The morphology of identified neuronal groups (i.e., parvalbumin, calbindin, and neuropeptide Y immunoreactive) in the echidna striatum and pallidum showed many similarities to those seen in eutherians, although the pattern of distribution of calbindin immunoreactive neurons was more uniform in the caudatoputamen of the echidna than in therians. These observations indicate that the same broad features of striatal and pallidal organization apply across all mammals and suggest that these common features may have arisen before the divergence of the monotreme and therian lineages. PMID:18821282

  10. Biology of Treponema pallidum: Correlation of Functional Activities With Genome Sequence Data

    Microsoft Academic Search

    Steven J. Norris; David L. Cox; George M. Weinstock

    Aspects of the biology of T. pallidum subsp. pallidum, the agent of syphilis, are examined in the context of a century of experimental studies and the recently determined genome sequence. T. pallidum and a group of closely related pathogenic spirochetes have evolved to become highly invasive, persistent pathogens with little toxigenic activity and an inability to survive outside the mammalian

  11. Genetic diversity in Treponema pallidum: implications for pathogenesis, evolution and molecular diagnostics of syphilis and yaws

    PubMed Central

    Šmajs, David; Norris, Steven J.; Weinstock, George M.

    2013-01-01

    Pathogenic uncultivable treponemes, similar to syphilis-causing Treponema pallidum subspecies pallidum, include T. pallidum ssp. pertenue, T. pallidum ssp. endemicum and Treponema carateum, which cause yaws, bejel and pinta, respectively. Genetic analyses of these pathogens revealed striking similarity among these bacteria and also a high degree of similarity to the rabbit pathogen, T. paraluiscuniculi, a treponeme not infectious to humans. Genome comparisons between pallidum and non-pallidum treponemes revealed genes with potential involvement in human infectivity, whereas comparisons between pallidum and pertenue treponemes identified genes possibly involved in the high invasivity of syphilis treponemes. Genetic variability within syphilis strains is considered as the basis of syphilis molecular epidemiology with potential to detect more virulent strains, whereas genetic variability within a single strain is related to its ability to elude the immune system of the host. Genome analyses also shed light on treponemal evolution and on chromosomal targets for molecular diagnostics of treponemal infections. PMID:22198325

  12. Genetic diversity in Treponema pallidum: implications for pathogenesis, evolution and molecular diagnostics of syphilis and yaws.

    PubMed

    Smajs, David; Norris, Steven J; Weinstock, George M

    2012-03-01

    Pathogenic uncultivable treponemes, similar to syphilis-causing Treponema pallidum subspecies pallidum, include T. pallidum ssp. pertenue, T. pallidum ssp. endemicum and Treponema carateum, which cause yaws, bejel and pinta, respectively. Genetic analyses of these pathogens revealed striking similarity among these bacteria and also a high degree of similarity to the rabbit pathogen, Treponema paraluiscuniculi, a treponeme not infectious to humans. Genome comparisons between pallidum and non-pallidum treponemes revealed genes with potential involvement in human infectivity, whereas comparisons between pallidum and pertenue treponemes identified genes possibly involved in the high invasivity of syphilis treponemes. Genetic variability within syphilis strains is considered as the basis of syphilis molecular epidemiology with potential to detect more virulent strains, whereas genetic variability within a single strain is related to its ability to elude the immune system of the host. Genome analyses also shed light on treponemal evolution and on chromosomal targets for molecular diagnostics of treponemal infections. PMID:22198325

  13. Assessment of the immune responses to Treponema pallidum Gpd DNA vaccine adjuvanted with IL-2 and chitosan nanoparticles before and after Treponema pallidum challenge in rabbits.

    PubMed

    Zhao, Feijun; Zhang, Xiaohong; Liu, Shuangquan; Zeng, Tiebing; Yu, Jian; Gu, Weiming; Zhang, Yuejun; Chen, Xi; Wu, Yimou

    2013-02-01

    Syphilis is a multistage, sexually transmitted disease caused by the spirochete, Treponema pallidum (Tp). A significantly high incidence of syphilis has been reported in several countries, including China, and there is an urgent need for the development of efficacious vaccines against syphilis. DNA vaccines are a major breakthrough in the field of vaccination with several advantages over traditional vaccines. Animal model studies of Tp DNA vaccines have not been reported elsewhere but our previous reports describe the development of a single-gene Tp DNA vaccine and preclinical immunization study. In this study, chitosan (CS) nanoparticles were used as a vector and an interleukin-2 expression plasmid (pIL-2) as an adjuvant to enhance a TpGpd DNA vaccine candidate (pTpGpd) in a rabbit Tp skin challenge model. At week 8 after the first immunization, three rabbits from each group were used to determine cytokine measurements and spleen lymphocyte proliferation assay. pTpGpd in combination with pIL-2 wrapped with CS led to the greatest enhancement of anti-TpGpd antibodies and T-cell proliferation. During infection, levels of anti-TpGpd antibodies and T-cell proliferation were measured. Both the serum special IgG and IL-2, interferon-? were significantly increased by the co-injection of the IL-2 plasmid compared with the injection of TpGpd DNA alone (P<0.05). Furthermore, IL-2 plasmid coinjection efficiently enhanced the antigen-specific lymphocyte proliferation response. Additionally, the ratios of positive skin lesions and ulcer lesions in groups immunized with pTpGpd were significantly lower than those of the pIL-2, CS or pIL-2 mixed with CS control groups (P<0.001). CS vectored and pIL-2 adjuvanted pTpGpd immunized animals exhibited the lowest rates of positive skin tests (8.33%) and ulcer lesions (4.17%) and the fastest recovery (42 d). These experiments indicate that co-injection of a pIL-2 plasmid with pTpGpd DNA vaccine wrapped with CS can significantly strengthen the long-term stability of immune response during infection, efficiently improve the protective effect against T. pallidum spirochetes infection and attenuate syphilitic lesion development. PMID:23334700

  14. The hyaluronidase associated with Treponema pallidum facilitates treponemal dissemination.

    PubMed

    Fitzgerald, T J; Repesh, L A

    1987-05-01

    Treponema pallidum contains hyaluronidase (Hase) associated with its surface. Experiments were performed to determine the functional role of this enzyme in syphilitic infection. The effects of incubating organisms with rabbit anti-bovine Hase or normal or immune sera were compared. Preincubation of treponemes with anti-Hase resulted in inhibition of treponemal degradation of hyaluronic acid, indicating that these antisera did in fact retard enzyme activity. Anti-Hase did not immobilize or neutralize T. pallidum. In addition, rabbits were immunized with bovine Hase and then challenged intradermally with organisms; subsequent lesion development was not affected. Anti-Hase did not block treponemal attachment to cultured testicular fibroblasts but did inhibit attachment to isolated capillaries. Rabbit amnions were used as an in vitro model for dissemination of T. pallidum. Anti-Hase retarded the penetration of organisms through the amnions. This inhibitory effect was dependent on the presence of amniotic hyaluronic acid. When this glycosaminoglycan was selectively removed, the anti-Hase lost its ability to inhibit treponemal penetration. When exogenous hyaluronic acid was added back to treated amnions, the inhibitory effect of anti-Hase was restored. Evans blue experiments were used to characterize treponeme-induced vascular leakage following intradermal inoculation of T. pallidum. Prior treatment of organisms with anti-Hase reduced dermal leakage of the dye, indicating the involvement of the treponemal Hase in causing vessel leakage. Finally, rabbit testicular infections were used as an in vivo model for dissemination; one testis was infected, and after 10 to 13 days, treponemes in the opposite testis were quantitated. The anti-Hase restricted dissemination of organisms. These findings point to the functional role of the treponemal Hase in facilitating disseminated syphilis. PMID:3552982

  15. The ventral pallidum and orbitofrontal cortex support food pleasantness inferences

    PubMed Central

    Simmons, W. Kyle; Rapuano, Kristina M.; Ingeholm, John E.; Avery, Jason; Kallman, Seth; Hall, Kevin D.; Martin, Alex

    2013-01-01

    Food advertisements often promote choices that are driven by inferences about the hedonic pleasures of eating a particular food. Given the individual and public health consequences of obesity, it is critical to address unanswered questions about the specific neural systems underlying these hedonic inferences. For example, although regions such as the orbitofrontal cortex (OFC) are frequently observed to respond more to pleasant food images than less hedonically pleasing stimuli, one important hedonic brain region in particular has largely remained conspicuously absent among human studies of hedonic response to food images. Based on rodent research demonstrating that activity in the ventral pallidum underlies the hedonic pleasures experienced upon eating food rewards, one might expect that activity in this important ‘hedonic hotspot’ might also track inferred food pleasantness. To date, however, no human studies have assessed this question. We thus asked human subjects to undergo fMRI and make item-by-item ratings of how pleasant it would be to eat particular visually perceived foods. Activity in the ventral pallidum was strongly modulated with pleasantness inferences. Additionally, activity within a region of the orbitofrontal cortex that tracks the pleasantness of tastes was also modulated with inferred pleasantness. Importantly, the reliability of these findings is demonstrated by their replication when we repeated the experiment at a new site with new subjects. These two experiments demonstrate that the ventral pallidum, in addition to the OFC, plays a central role in the moment-to-moment hedonic inferences that influence food-related decision-making. PMID:23397317

  16. A redescripton of Lyrosoma pallidum (Eschscholtz) and distributional range extension of Lyrosoma Mannerheim (Coleoptera, Agyrtidae)

    PubMed Central

    Yoo, In-Seong; Sikes, Derek; Ahn, Kee-Jeong

    2013-01-01

    Abstract A redescription with illustrations of the species Lyrosoma pallidum and a key to the Korean species of the family Agyrtidae are provided. New distributional data, including a range extension, of the two Lyrosoma Mannerheim species are presented. Lyrosoma pallidum (Eschscholtz) is recorded for the first time in Korea. PMID:24146551

  17. Detection of Nonspecific Resistance to Listeria monocytogenes in Rabbits Infected with Treponema pallidum

    PubMed Central

    Schell, Ronald F.; Musher, Daniel M.

    1974-01-01

    Several lines of evidence suggest that cell-mediated immunity (CMI) is suppressed in the early stages of infection caused by Treponema pallidum and becomes activated at the time that latency is induced. In the studies reported in this paper, rabbits were infected intravenously with T. pallidum and subsequently challenged with Listeria monocytogenes. Enhanced ability to suppress the growth of Listeria was detected in their livers between 3 and 5 weeks after infection with T. pallidum, corresponding to the onset and regression of the generalized syphilitic eruption. A second infection of T. pallidum 4 weeks after the first, at a time when suppression was beginning to wane, prolonged the listericidal activity. These observations support the hypothesis that infection by T. pallidum stimulates CMI, which, in turn, may play a role in inducing latency. PMID:4207188

  18. TprC/D (Tp0117/131), a Trimeric, Pore-Forming Rare Outer Membrane Protein of Treponema pallidum, Has a Bipartite Domain Structure

    PubMed Central

    Anand, Arvind; Luthra, Amit; Dunham-Ems, Star; Caimano, Melissa J.; Karanian, Carson; LeDoyt, Morgan; Cruz, Adriana R.; Salazar, Juan C.

    2012-01-01

    Identification of Treponema pallidum rare outer membrane proteins (OMPs) has been a longstanding objective of syphilis researchers. We recently developed a consensus computational framework that employs a battery of cellular localization and topological prediction tools to generate ranked clusters of candidate rare OMPs (D. L. Cox et al., Infect. Immun. 78:5178–5194, 2010). TP0117/TP0131 (TprC/D), a member of the T. pallidum repeat (Tpr) family, was a highly ranked candidate. Circular dichroism, heat modifiability by SDS-PAGE, Triton X-114 phase partitioning, and liposome incorporation confirmed that full-length, recombinant TprC (TprCFl) forms a ?-barrel capable of integrating into lipid bilayers. Moreover, TprCFl increased efflux of terbium-dipicolinic acid complex from large unilamellar vesicles and migrated as a trimer by blue-native PAGE. We found that in T. pallidum, TprC is heat modifiable, trimeric, expressed in low abundance, and, based on proteinase K accessibility and opsonophagocytosis assays, surface exposed. From these collective data, we conclude that TprC is a bona fide rare OMP as well as a functional ortholog of Escherichia coli OmpF. We also discovered that TprC has a bipartite architecture consisting of a soluble N-terminal portion (TprCN), presumably periplasmic and bound directly or indirectly to peptidoglycan, and a C-terminal ?-barrel (TprCC). Syphilitic rabbits generate antibodies exclusively against TprCC, while secondary syphilis patients fail to mount a detectable antibody response against either domain. The syphilis spirochete appears to have resolved a fundamental dilemma arising from its extracellular lifestyle, namely, how to enhance OM permeability without increasing its vulnerability to the antibody-mediated defenses of its natural human host. PMID:22389487

  19. Sensitive detection of Treponema pallidum by using the polymerase chain reaction.

    PubMed

    Burstain, J M; Grimprel, E; Lukehart, S A; Norgard, M V; Radolf, J D

    1991-01-01

    We have developed a sensitive assay for Treponema pallidum subsp. pallidum (T. pallidum), the agent of veneral syphilis, based upon the polymerase chain reaction (PCR). A 658-bp portion of the gene encoding the 47-kDa membrane immunogen was amplified, and the PCR products were probed by DNA-DNA hybridization with a 496-bp fragment internal to the amplitifed DNA. The assay detected approximately 0.01 pg of purified T. pallidum DNA, and positive results were obtained routinely from suspensions of treponemes calculated to contain 10 or more organism and from some suspensions calculated to contain a single organism. Specific PCR products were obtained for the closely related agent of yaws, Treponema pallidum subsp. pertenue, but not with human DNA or DNAs from other spirochetes (including Borrelia burgdoferi), skin microorganisms, sexually transmitted disease pathogens, and central nervous system pathogens. T. pallidum DNA was detected in serum, cerebrospinal fluids, and amniotic fluids from syphilis patients but not in in nonsyphilitic controls. T. pallidum DNA was also amplified from paraffin-embedded tissue. The diagnosis of syphillis by using PCR may become a significant addition to the diagnostic armamentarium and a valuable technique for the investigation of syphilis pathogenesis. PMID:1993770

  20. 21 CFR 866.3830 - Treponema pallidum tre-ponemal test reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...pallidum directly from infecting treponemal organisms in serum. The identification aids in the diagnosis of syphilis caused by bacteria belonging to the genus Treponema and provides epidemiological information on syphilis. (b) Classification....

  1. 21 CFR 866.3830 - Treponema pallidum tre-ponemal test reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3830 Treponema pallidum tre-ponemal test reagents. (a)...

  2. 21 CFR 866.3830 - Treponema pallidum tre-ponemal test reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3830 Treponema pallidum tre-ponemal test reagents. (a)...

  3. 21 CFR 866.3820 - Treponema pallidum non-treponemal test reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3820 Treponema pallidum non-treponemal test reagents. (a)...

  4. 21 CFR 866.3830 - Treponema pallidum tre-ponemal test reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3830 Treponema pallidum tre-ponemal test reagents. (a)...

  5. 21 CFR 866.3820 - Treponema pallidum non-treponemal test reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3820 Treponema pallidum non-treponemal test reagents. (a)...

  6. Limited protection of rabbits against infection with Treponema pallidum by immune rabbit sera.

    PubMed Central

    Graves, S; Alden, J

    1979-01-01

    After intradermal infection of rabbits with 3 x 10(6) Treponema pallidum (Melbourne 1 strain) samples of serum were taken at one, two, three, four, and six months after infection. Normal rabbits were passively immunised with these sera, challenged with intradermal doses (10(4), 10(3), 10(2), 10) of T. pallidum, and the latent periods of infection, lesion diameters, and the number of inoculation sites developing into lesions were observed. The sera taken at three, four, and six months reduced the number of intradermal inoculation sites that developed into syphilitic lesions after challenge with 10 T. pallidum. These same three sera also increased the latent period of infection after challenge with 10(4) T. pallidum. The transfer of 50 ml of immune serum per rabbit over a nine-day period before challenge had very little effect on the course of the challenge infection. Only a low level of immunity in rabbits to this strain of T. pallidum appears to be mediated by immune serum but this small degree of protection did increase with time after infection. Enhanced growth of T. pallidum in the serum-recipient rabbits did not occur, thus suggesting that none of the sera was immunosuppressive. PMID:393362

  7. Failure of PCR to Detect Treponema pallidum ssp. pertenue DNA in Blood in Latent Yaws

    PubMed Central

    Chi, Kai-Hua; Vahi, Ventis; Sun, Yongcheng; Mabey, David C.; Solomon, Anthony W.; Chen, Cheng Y.; Pillay, Allan

    2015-01-01

    Yaws, caused by Treponema pallidum ssp. pertenue, is a neglected tropical disease closely related to venereal syphilis and is targeted for eradication by 2020. Latent yaws represents a diagnostic challenge, and current tools cannot adequately distinguish between individuals with true latent infection and individuals who are serofast following successful treatment. PCR on blood has previously been shown to detect T. pallidum DNA in patients with syphilis, suggesting that this approach may be of value in yaws. We performed real-time PCR for Treponema pallidum ssp. pertenue on blood samples from 140 children with positive T. pallidum Particle Agglutination (TPPA) and Rapid Plasma Reagin (RPR) tests and 7 controls (negative serology), all collected as part of a prospective study of yaws in the Solomon Islands. All samples were also tested by a nested PCR for T. pallidum. 12 patients had clinical evidence of active yaws whilst 128 were considered to have latent yaws. 43 children had high titre rapid plasma reagins (RPRs) of ?1:32. PCR testing with both assays gave negative results in all cases. It is possible that the failure to detect T. pallidum ssp. pertenue in blood reflects lower loads of organism in latent yaws compared to those in latent infection with T. pallidum ssp. pertenue, and/or a lower propensity for haematogenous dissemination in yaws than in syphilis. As the goal of the yaws control programme is eradication, a tool that can differentiate true latent infection from individuals who are serofast would be of value; however, PCR of blood is not that tool. PMID:26125585

  8. Failure of PCR to Detect Treponema pallidum ssp. pertenue DNA in Blood in Latent Yaws.

    PubMed

    Marks, Michael; Katz, Samantha; Chi, Kai-Hua; Vahi, Ventis; Sun, Yongcheng; Mabey, David C; Solomon, Anthony W; Chen, Cheng Y; Pillay, Allan

    2015-06-01

    Yaws, caused by Treponema pallidum ssp. pertenue, is a neglected tropical disease closely related to venereal syphilis and is targeted for eradication by 2020. Latent yaws represents a diagnostic challenge, and current tools cannot adequately distinguish between individuals with true latent infection and individuals who are serofast following successful treatment. PCR on blood has previously been shown to detect T. pallidum DNA in patients with syphilis, suggesting that this approach may be of value in yaws. We performed real-time PCR for Treponema pallidum ssp. pertenue on blood samples from 140 children with positive T. pallidum Particle Agglutination (TPPA) and Rapid Plasma Reagin (RPR) tests and 7 controls (negative serology), all collected as part of a prospective study of yaws in the Solomon Islands. All samples were also tested by a nested PCR for T. pallidum. 12 patients had clinical evidence of active yaws whilst 128 were considered to have latent yaws. 43 children had high titre rapid plasma reagins (RPRs) of ?1:32. PCR testing with both assays gave negative results in all cases. It is possible that the failure to detect T. pallidum ssp. pertenue in blood reflects lower loads of organism in latent yaws compared to those in latent infection with T. pallidum ssp. pertenue, and/or a lower propensity for haematogenous dissemination in yaws than in syphilis. As the goal of the yaws control programme is eradication, a tool that can differentiate true latent infection from individuals who are serofast would be of value; however, PCR of blood is not that tool. PMID:26125585

  9. Genome analysis of Treponema pallidum subsp. pallidum and subsp. pertenue strains: most of the genetic differences are localized in six regions.

    PubMed

    Mikalová, Lenka; Strouhal, Michal; ?ejková, Darina; Zobaníková, Marie; Pospíšilová, Petra; Norris, Steven J; Sodergren, Erica; Weinstock, George M; Šmajs, David

    2010-01-01

    The genomes of eight treponemes including T. p. pallidum strains (Nichols, SS14, DAL-1 and Mexico A), T. p. pertenue strains (Samoa D, CDC-2 and Gauthier), and the Fribourg-Blanc isolate, were amplified in 133 overlapping amplicons, and the restriction patterns of these fragments were compared. The approximate sizes of the genomes investigated based on this whole genome fingerprinting (WGF) analysis ranged from 1139.3-1140.4 kb, with the estimated genome sequence identity of 99.57-99.98% in the homologous genome regions. Restriction target site analysis, detecting the presence of 1773 individual restriction sites found in the reference Nichols genome, revealed a high genome structure similarity of all strains. The unclassified simian Fribourg-Blanc isolate was more closely related to T. p. pertenue than to T. p. pallidum strains. Most of the genetic differences between T. p. pallidum and T. p. pertenue strains were accumulated in six genomic regions. These genome differences likely contribute to the observed differences in pathogenicity between T. p. pallidum and T. p. pertenue strains. These regions of sequence divergence could be used for the molecular detection and discrimination of syphilis and yaws strains. PMID:21209953

  10. Yaws: 110 Years After Castellani's Discovery of Treponema pallidum subspecies pertenue.

    PubMed

    Stamm, Lola V

    2015-07-01

    Yaws is a neglected infectious disease that affects mostly children and adolescents living in poor, rural communities in humid, tropical areas of Africa, southeast Asia, and the Pacific Islands. The etiological agent of yaws, Treponema pallidum subspecies pertenue (T. pertenue), was discovered by Aldo Castellani in 1905 shortly after Schaudinn and Hoffmann discovered the etiological agent of syphilis, T. pallidum subspecies pallidum. The discovery of T. pertenue enabled the development of animal models and the identification of an effective antibiotic treatment (i.e., penicillin) for yaws. A World Health Organization (WHO) mass treatment campaign from 1952 to 1964 reduced the global burden of yaws by 95%, but failed to eradicate this disease. Today, 110 years after Castellani's discovery of T. pertenue, yaws is again targeted for eradication. Recent advances in the treatment and diagnosis of yaws improve the likelihood of success this time. However, several challenges must be overcome to make the goal of yaws eradication attainable. PMID:25870417

  11. Membrane topology and cellular location of the Treponema pallidum glycerophosphodiester phosphodiesterase (GlpQ) ortholog.

    PubMed

    Shevchenko, D V; Sellati, T J; Cox, D L; Shevchenko, O V; Robinson, E J; Radolf, J D

    1999-05-01

    Recent reports that isolated Treponema pallidum outer membranes contain an ortholog for glycerophosphodiester phosphodiesterase (GlpQ) (D. V. Shevchenko, D. R. Akins, E. J. Robinson, M. Li, O. V. Shevchenko, and J. D. Radolf, Infect. Immun. 65:4179-4189, 1997) and that this protein is a potential opsonic target for T. pallidum (C. E. Stebeck, J. M. Shaffer, T. W. Arroll, S. A. Lukehart, and W. C. Van Voorhis, FEMS Microbiol. Lett. 154:303-310, 1997) prompted a more detailed investigation of its physicochemical properties and cellular location. [14C]palmitate radiolabeling studies of a GlpQ-alkaline phosphatase fusion expressed in Escherichia coli confirmed the prediction from DNA sequencing that the protein is lipid modified. Studies using Triton X-114 phase partitioning revealed that the protein's amphiphilicity is due to lipid modification and that a substantial portion of the polypeptide is associated with the T. pallidum peptidoglycan sacculus. Three different approaches, i.e., (i) proteinase K treatment of intact treponemes, (ii) indirect immunofluorescence analysis of treponemes encapsulated in agarose beads, and (iii) opsonophagocytosis of treponemes incubated with antiserum against recombinant GlpQ by rabbit peritoneal macrophages, confirmed that GlpQ is entirely subsurface in T. pallidum. Moreover, rabbits hyperimmunized with GlpQ were not protected against intradermal challenge with virulent treponemes. Circular dichroism spectroscopy confirmed that the recombinant form of the polypeptide lacked discernible evidence of denaturation. Finally, GlpQ was not radiolabeled when T. pallidum outer membranes were incubated with 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)-diazarene, a photoactivatable, lipophilic probe which promiscuously labels both proteins and lipids within phospholipid bilayers. Taken as a whole, these studies indicate that the T. pallidum GlpQ ortholog is a periplasmic protein associated predominantly with the spirochete's peptidoglycan-cytoplasmic membrane complex. PMID:10225883

  12. Retention of motility and virulence of Treponema pallidum (Nichols strain) in vitro.

    PubMed Central

    Graves, S R; Sandok, P L; Jenkin, H M; Johnson, R C

    1975-01-01

    A maintenance medium for Treponema pallidum was designed to hold its Eh at the optimum for that organism, -10 to -110 mV. After 100% motile (freshly harvested) T. pallidum was inoculated into the medium, the motility of the treponemes decreased to 80% after 2 days, 50% after 3.5 days, and 0% after 9 days during incubation at 34 C. Full virulence was retained for 2 days, but it dropped rapidly thereafter, and the treponemes became avirulent by day 5. PMID:1104483

  13. Protective efficacy of a Treponema pallidum Gpd DNA vaccine vectored by chitosan nanoparticles and fused with interleukin-2.

    PubMed

    Zhao, Feijun; Wang, Shiping; Zhang, Xiaohong; Gu, Weiming; Yu, Jian; Liu, Shuangquan; Zeng, Tiebing; Zhang, Yuejun; Wu, Yimou

    2012-02-01

    In the present study, immunomodulatory responses of a DNA vaccine constructed by fusing Treponema pallidum (Tp) glycerophosphodiester phosphodiesterase (Gpd) to interleukin-2 (IL-2) and using chitosan (CS) nanoparticles as vectors were investigated. New Zealand white rabbits were immunized by intramuscular inoculation of control DNAs, Tp Gpd DNA vaccine, or Gpd-IL-2 fusion DNA vaccine, which were vectored by CS nanoparticles. Levels of the anti-Gpd antibodies and levels of IL-2 and interferon-? in rabbits were increased upon inoculation of Gpd-IL-2 fusion DNA vaccine, when compared with the inoculation with Gpd DNA vaccine, with CS vectoring increasing the effects. The Gpd-IL-2 fusion DNA vaccine efficiently enhanced the antigen-specific lymphocyte proliferative response. When the rabbits were challenged intradermally with 10(5) Tp (Nichols) spirochetes, the Gpd-IL-2 fusion DNA vaccine conferred better protection than the Gpd DNA vaccine (P < 0.05), as characterized by lower detectable amounts of dark field positive lesions (17.5%), lower ulcerative lesion scores (15%), and faster recovery. Individuals treated with the Tp Gpd-IL-2 fusion DNA vaccine vectored by CS nanoparticles had the lowest amounts of dark field positive lesions (10%) and ulcerations (5%) observed and the fastest recovery (42 days). These results indicate that the Gpd-IL-2 fusion DNA vaccine vectored by CS nanoparticles can efficiently induce Th1-dominant immune responses, improve protective efficacy against Tp spirochete infection, and effectively attenuate development of syphilitic lesions. PMID:22260167

  14. Treponema pallidum putative novel drug target identification and validation: rethinking syphilis therapeutics with plant-derived terpenoids.

    PubMed

    Dwivedi, Upendra N; Tiwari, Sameeksha; Singh, Priyanka; Singh, Swati; Awasthi, Manika; Pandey, Veda P

    2015-02-01

    Syphilis, a slow progressive and the third most common sexually transmitted disease found worldwide, is caused by a spirochete gram negative bacteria Treponema pallidum. Emergence of antibiotic resistant T. pallidum has led to a search for novel drugs and their targets. Subtractive genomics analyses of pathogen T. pallidum and host Homo sapiens resulted in identification of 126 proteins essential for survival and viability of the pathogen. Metabolic pathway analyses of these essential proteins led to discovery of nineteen proteins distributed among six metabolic pathways unique to T. pallidum. One hundred plant-derived terpenoids, as potential therapeutic molecules against T. pallidum, were screened for their drug likeness and ADMET (absorption, distribution, metabolism, and toxicity) properties. Subsequently the resulting nine terpenoids were docked with five unique T. pallidum targets through molecular modeling approaches. Out of five targets analyzed, D-alanine:D-alanine ligase was found to be the most promising target, while terpenoid salvicine was the most potent inhibitor. A comparison of the inhibitory potential of the best docked readily available natural compound, namely pomiferin (flavonoid) with that of the best docked terpenoid salvicine, revealed that salvicine was a more potent inhibitor than that of pomiferin. To the best of our knowledge, this is the first report of a terpenoid as a potential therapeutic molecule against T. pallidum with D-alanine:D-alanine ligase as a novel target. Further studies are warranted to evaluate and explore the potential clinical ramifications of these findings in relation to syphilis that has public health importance worldwide. PMID:25683888

  15. The immune response to infection with Treponema pallidum, the stealth pathogen

    Microsoft Academic Search

    Juan C. Salazar; Karsten R. O. Hazlett; Justin D. Radolf

    2002-01-01

    Cutaneous immunobiology and spirochetal molecular biology have allowed investigators to propose a conceptual framework for the development of both the innate and adaptive immune response to Treponema pallidum infection. While some clinical manifestations can be attributed to humoral responses, most can be attributed to a combination of local innate and adaptive cellular immunity.

  16. Use of Treponema pallidum PCR in Testing of Ulcers for Diagnosis of Primary Syphilis1

    PubMed Central

    Sednaoui, Patrice; Lautenschlager, Stephan; Ferry, Tristan; Toutous-Trellu, Laurence; Cavassini, Matthias; Yassir, Fatima; Martinez de Tejada, Begońa; Emonet, Stéphane; Combescure, Christophe; Schrenzel, Jacques; Perneger, Thomas

    2015-01-01

    Treponema pallidum PCR (Tp-PCR) has been noted as a valid method for diagnosing syphilis. We compared Tp-PCR to a combination of darkfield microscopy (DFM), the reference method, and serologic testing in a cohort of 273 patients from France and Switzerland and found the diagnostic accuracy of Tp-PCR was higher than that for DFM. PMID:25531672

  17. MOLECULAR CHARACTERIZATION OF RECEPTOR BINDING PROTEINS AND IMMUNOGENS OF VIRULENT TREPONEMA PALLIDUM

    Microsoft Academic Search

    JOEL B. BASEMAN; EDWARD C. HAYES

    During the clinical course of syphilis, a complex interrelationship exists between virulent Treponema pallidum and the parasitized host. Infection can persist in the presence of a significant immune response, and manifestations of actively developing disease are well documented (1, 2). Although controversies still remain concei'ning the relative contributions of cellular and humoral immunity to eradication of the disease, it is

  18. Defining the Interaction of the Treponema pallidum Adhesin Tp0751 with Laminin

    Microsoft Academic Search

    Caroline E. Cameron; Nathan L. Brouwer; Lisa M. Tisch; Janelle M. Y. Kuroiwa

    2005-01-01

    Various invasive pathogens attach to host tissues via the extracellular matrix component laminin, the major glycoprotein found within basement membranes. Previous investigations identified the laminin-binding adhe- sin Tp0751 within the spirochete bacterium Treponema pallidum. In the current study, Tp0751 was shown to attach to a variety of laminin isoforms that are widely distributed throughout the host, including laminins 1, 2,

  19. The genome of Treponema pallidum: new light on the agent of syphilis.

    PubMed

    Weinstock, G M; Hardham, J M; McLeod, M P; Sodergren, E J; Norris, S J

    1998-10-01

    Treponema pallidum subsp, pallidum, the causative agent of the sexually transmitted disease syphilis, is a fastidious, microaerophilic obligate parasite of humans. This bacterium is one of the few prominent infectious agents that has not been cultured continuously in vitro and consequently relatively little is known about its virulence mechanisms at the molecular level. T. pallidum therefore represented an attractive candidate for genomic sequencing. The complete genome sequence of T. pallidum has now been completed and comprises 1,138,006 base pairs containing 1041 predicted protein coding sequences. An important goal of this project is to identify possible virulence factors. Analysis of the genome indicates a number of potential virulence factors including a family of 12 proteins related to the Msp protein of Treponema denticola, a number of putative hemolysins, as well as several other classes of proteins of interest. The results of this analysis are reviewed in this article and indicate the value of whole genome sequences for rapidly advancing knowledge of infectious agents. PMID:9862125

  20. Treponema pallidum infection in the wild baboons of East Africa: distribution and genetic characterization of the strains responsible.

    PubMed

    Harper, Kristin N; Fyumagwa, Robert D; Hoare, Richard; Wambura, Philemon N; Coppenhaver, Dorian H; Sapolsky, Robert M; Alberts, Susan C; Tung, Jenny; Rogers, Jeffrey; Kilewo, Morris; Batamuzi, Emmanuel K; Leendertz, Fabian H; Armelagos, George J; Knauf, Sascha

    2012-01-01

    It has been known for decades that wild baboons are naturally infected with Treponema pallidum, the bacterium that causes the diseases syphilis (subsp. pallidum), yaws (subsp. pertenue), and bejel (subsp. endemicum) in humans. Recently, a form of T. pallidum infection associated with severe genital lesions has been described in wild baboons at Lake Manyara National Park in Tanzania. In this study, we investigated ten additional sites in Tanzania and Kenya using a combination of macroscopic observation and serology, in order to determine whether the infection was present in each area. In addition, we obtained genetic sequence data from six polymorphic regions using T. pallidum strains collected from baboons at two different Tanzanian sites. We report that lesions consistent with T. pallidum infection were present at four of the five Tanzanian sites examined, and serology was used to confirm treponemal infection at three of these. By contrast, no signs of treponemal infection were observed at the six Kenyan sites, and serology indicated T. pallidum was present at only one of them. A survey of sexually mature baboons at Lake Manyara National Park in 2006 carried out as part of this study indicated that roughly ten percent displayed T. pallidum-associated lesions severe enough to cause major structural damage to the genitalia. Finally, we found that T. pallidum strains from Lake Manyara National Park and Serengeti National Park were genetically distinct, and a phylogeny suggested that baboon strains may have diverged prior to the clade containing human strains. We conclude that T. pallidum infection associated with genital lesions appears to be common in the wild baboons of the regions studied in Tanzania. Further study is needed to elucidate the infection's transmission mode, its associated morbidity and mortality, and the relationship between baboon and human strains. PMID:23284649

  1. Evaluation of the recombinant protein TpF1 of Treponema pallidum for serodiagnosis of syphilis.

    PubMed

    Jiang, Chuanhao; Zhao, Feijun; Xiao, Jinhong; Zeng, Tiebing; Yu, Jian; Ma, Xiaohua; Wu, Haiying; Wu, Yimou

    2013-10-01

    Syphilis is a chronic infection caused by Treponema pallidum subsp. pallidum, and diagnosis with sensitive and specific methods is a challenging process that is important for its prevention and treatment. In the present study, we established a recombinant protein TpF1-based indirect immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) and a Western blot assay for human and rabbit sera. The 20-kDa recombinant protein TpF1 was detected by Western blotting performed with sera from rabbits immunized with recombinant TpF1 and infected with the T. pallidum Nichols strain and T. pallidum clinical isolates but was not detected by Western blotting with sera from uninfected rabbits. The sensitivity of the recombinant protein was determined by screening sera from individuals with primary, secondary, latent, and congenital syphilis (n = 82). The specificity of the recombinant protein was determined by screening sera from uninfected controls (n = 30) and individuals with potentially cross-reactive infections, including Lyme disease (n = 30) and leptospirosis (n = 5). The sensitivities of TpF1-based ELISAs were 93.3%, 100%, 100%, and 100% for primary, secondary, latent, and congenital syphilis, respectively, and the specificities were all 100% for sera from uninfected controls and individuals with potentially cross-reactive infections. In Western blot assays, the sensitivities and specificities of TpF1 for human sera were all 100%. The reactivities of TpF1 with syphilitic sera were proportional to the titers of the T. pallidum particle agglutination (TPPA) assay. These data indicate that the recombinant protein TpF1 is a highly immunogenic protein in human and rabbit infections and a promising marker for the screening of syphilis. PMID:23945159

  2. CpG adjuvant enhances the mucosal immunogenicity and efficacy of a Treponema pallidum DNA vaccine in rabbits

    PubMed Central

    Zhao, Feijun; Liu, Shuangquan; Zhang, Xiaohong; Yu, Jian; Zeng, Tiebing; Gu, Weiming; Cao, Xunyu; Chen, Xi; Wu, Yimou

    2013-01-01

    Objectives: The protective response against Treponema pallidum (Tp) infection of a DNA vaccine enhanced by an adjuvant CpG ODN was investigated. Results: The mucosal adjuvant CpG ODN enhanced the production of higher levels of anti-TpGpd antibodies induced by pcD/Gpd-IL-2 in rabbits. It also resulted in higher levels of secretion of IL-2 and IFN-?, and facilitated T cell proliferation and differentiation (p < 0.05). No significant difference about testing index above-mentioned was found in the intranasal immunization group of pcD/Gpd-IL-2 vaccine adjuvanted by CpG ODN when compared with the immunization by pcD/Gpd-IL-2 vaccine intramuscular injection alone (p > 0.05). Furthermore, CpG ODN stimulated the production of mucosa-specific anti-sIgA antibodies and resulted in the lowest Tp-positive rate (6.7%) for Tp-infection of skin lesions and the lowest rates (8.3%) of ulceration lesions, thus achieving better protective effects. Methods: New Zealand rabbits were immunized with the eukaryotic vector encoding recombinant pcD/Gpd-IL-2 using intramuscular multi-injection or together with mucosal enhancement via a nasal route. The effect of the mucosal adjuvant CpG ODN was examined. Conclusions:The CpG ODN adjuvant significantly enhances the humoral and cellular immune effects of the immunization by pcD/Gpd-IL-2 with mucosal enhancement via nasal route. It also stimulates strong mucosal immune effects, thus initiating more efficient immune-protective effects. PMID:23563515

  3. Whole Genome Sequences of Three Treponema pallidum ssp. pertenue Strains: Yaws and Syphilis Treponemes Differ in Less than 0.2% of the Genome Sequence

    Microsoft Academic Search

    Darina ?ejková; Marie Zobaníková; Lei Chen; Petra Pospíšilová; Michal Strouhal; Xiang Qin; Lenka Mikalová; Steven J. Norris; Donna M. Muzny; Richard A. Gibbs; Lucinda L. Fulton; Erica Sodergren; George M. Weinstock; David Šmajs

    2012-01-01

    BackgroundThe yaws treponemes, Treponema pallidum ssp. pertenue (TPE) strains, are closely related to syphilis causing strains of Treponema pallidum ssp. pallidum (TPA). Both yaws and syphilis are distinguished on the basis of epidemiological characteristics, clinical symptoms, and several genetic signatures of the corresponding causative agents.Methodology\\/Principal FindingsTo precisely define genetic differences between TPA and TPE, high-quality whole genome sequences of three

  4. Radiolabeling of Treponema pallidum (Nichols virulent strain) in vitro with precursors for protein and RNA biosynthesis.

    PubMed Central

    Sandok, P L; Jenkin, H M

    1978-01-01

    We observed uptake of [U-14C]serine, U-14C-labeled amino acid hydrolysates, and [2-14C]uracil by virulent Treponema pallidum in vitro for at least 96 h. No uptake of [2-14C]thymine, [1-14C]pyruvate, [U-14C]pyruvate, and [2-14C]uridine was detected. Treponemal protein and RNA biosynthetic activity was identified by erythromycin inhibition of amino acid and uracil uptake. Radioactivity due to uptake of radiolabeled amino acids by residual testicular cells in the cultures remained at background levels regardless of the presence or absence of cycloheximide. Accumulation of the radiolabeled substrates by T. pallidum proceeded at a linear rate for 48 to 96 h during incubation in vitro. The longevity of substrate uptake using the system of incubation described will facilitate future studies on the metabolism of the microorganism to help determine essential growth factors and environmental conditions for multiplication of T. pallidum in vitro. PMID:365745

  5. Identification and characterization of the Treponema pallidum tpn50 gene, an ompA homolog.

    PubMed Central

    Hardham, J M; Stamm, L V

    1994-01-01

    Treponema pallidum is a pathogenic spirochete that has no known genetic exchange mechanisms. In order to identify treponemal genes encoding surface and secreted proteins, we carried out TnphoA mutagenesis of a T. pallidum genomic DNA library in Escherichia coli. Several of the resulting clones expressed enzymatically active T. pallidum-alkaline phosphatase fusion proteins. The DNA sequence of the 5' portion of a number of the treponemal genes was obtained and analyzed. A recombinant clone harboring plasmid p4A2 that encoded a treponemal protein with an approximate molecular mass of 50,000 Da was identified. Plasmid p4A2 contained an open reading frame of 1,251 nucleotides that resulted in a predicted protein of 417 amino acids with a calculated molecular mass of 47,582 Da. We have named this gene tpn50 in accordance with the current nomenclature for T. pallidum genes. A 1.9-kb HincII-ClaI fragment from p4A2 that contained the tpn50 gene was subcloned to produce p4A2HC2. Comparison of the predicted amino acid sequence of TpN50 with protein sequences in the National Center for Biotechnology Information data base indicated statistically significant homology to the Pseudomonas sp. OprF, E. coli OmpA, Bordetella avium OmpA, Neisseria meningitidis RmpM, Neisseria gonorrhoeae PIII, Haemophilus influenzae P6, E. coli PAL, and Legionella pneumophila PAL proteins. These proteins are all members of a family of outer membrane proteins that are present in gram-negative bacteria. The tpn50 gene complemented E. coli ompA mutations on the basis of two separate criteria. First, morphometry and electron microscopy data showed that E. coli C386 (ompA lpp) cells harboring plasmid vector pEBH21 were rounded while cells of the same strain harboring p4A2HC2 (TpN50+), pWW2200 (OprF+), or pRD87 (OmpA+) were rod shaped. Second, E. coli BRE51 (MC4100 delta sulA-ompA) cells harboring pEBH21 grew poorly at 42 degrees C in minimal medium, while the growth of BRE51 cells harboring p4A2HC2 was similar to that of the parental MC4100 cells. These results demonstrate that the TpN50 protein is functionally equivalent to the E. coli OmpA protein. If TpN50 functions in a similar fashion in T. pallidum, then it may be localized to the treponemal outer membrane. Images PMID:8112835

  6. Immunity in Experimental Syphilis IV. Serological Reactivity of Antigens Extracted from ?-Irradiated Treponema pallidum and Treponema reiteri

    PubMed Central

    Miller, James N.; De Bruijn, J. H.; Bekker, J. H.

    1966-01-01

    Miller, James N. (University of California School of Medicine, Los Angeles), J. H. De Bruijn, and J. H. Bekker. Immunity in experimental syphilis. IV. Serological reactivity of antigens extracted from ?-irradiated Treponema pallidum and Treponema reiteri. J. Bacteriol. 91:583–587. 1966.—Ultrasonic lysate preparations extracted from virulent Treponema pallidum, Nichols strain, suspensions exposed to 652,800 R of ?-irradiation exhibited a loss in the serological reactivity of their heat-labile antigens; the heat-stable components of both the lysate and residue antigens were unaffected. The activity of heat-stable, cardiolipin T. pallidum complement-fixing antigen obtained from similarly irradiated organisms was also unaltered. ?-Irradiation of the cultivable Treponema reiteri with dosages as high as 6,500,000 R failed to alter serologically either the heat-labile or heat-stable component of its lipopolysaccharide-protein (Reiter protein) antigen. The reactivity of the lipopolysaccharide portion of the Reiter protein complex with an antiserum to T. pallidum Nichols indicates previously unsuspected antigenic differences between the rabbit-adapted Nichols strain of the organism and so-called “wild” human strains of T. pallidum in which this antigen is generally absent. PMID:5327359

  7. Molecular differentiation of Treponema pallidum subspecies in skin ulceration clinically suspected as yaws in Vanuatu using real-time multiplex PCR and serological methods.

    PubMed

    Chi, Kai-Hua; Danavall, Damien; Taleo, Fasihah; Pillay, Allan; Ye, Tun; Nachamkin, Eli; Kool, Jacob L; Fegan, David; Asiedu, Kingsley; Vestergaard, Lasse S; Ballard, Ronald C; Chen, Cheng-Yen

    2015-01-01

    We developed a TaqMan-based real-time quadriplex polymerase chain reaction (PCR) to simultaneously detect Treponema pallidum subspecies pallidum, T. pallidum subsp. pertenue, and T. pallidum subsp. endemicum, the causative agents of venereal syphilis, yaws, and bejel, respectively. The PCR assay was applied to samples from skin ulcerations of clinically presumptive yaws cases among children on Tanna Island, Vanuatu. Another real-time triplex PCR was used to screen for the point mutations in the 23S rRNA genes that have previously been associated with azithromycin resistance in T. pallidum subsp. pallidum strains. Seropositivity by the classical syphilis serological tests was 35.5% among children with skin ulcerations clinically suspected with yaws, whereas the presence of T. pallidum subsp. pertenue DNA was only found in lesions from 15.5% of children. No evidence of T. pallidum subsp. pertenue infection, by either PCR or serology was found in ?59% of cases indicating alternative causes of yaws-like lesions in this endemic area. PMID:25404075

  8. Studies of rabbit testes infected with Treponema pallidum. III. Immunosuppressive activity of infiltrating mononuclear cells.

    PubMed Central

    Wicher, V; Wicher, K

    1984-01-01

    When mononuclear cells infiltrating rabbit testes infected with Treponema pallidum were cocultured with autologous or homologous peripheral blood lymphocytes spontaneous stimulation and that induced by concanavalin A were suppressed. The inhibition was not due to the cytotoxic effect of the mononuclear cells or to their interference with the active site of concanavalin A (competitive inhibition). The suppressor activity was present in both T and non-T cells but was not affected by pretreatment of the mononuclear cells with indomethacin, a prostaglandin synthetase inhibitor. The suppressor activity may be intrinsic to the mononuclear cells or acquired by the cells in the testicular environment. PMID:6230135

  9. Comparison of the morphology, antigenicity, and pathology of Nichols and Birmingham isolates of Treponema pallidum.

    PubMed

    Penn, C W; Clay, J C

    1982-10-01

    A strain of Treponema pallidum isolated from a homosexually acquired penile chancre in Birmingham in 1979 was serially passaged in rabbits and compared with the Nichols strain. The two strains appeared identical morphologically, and no gross differences were detected on antigenic analysis by two-dimensional immunoelectrophoresis. The Birmingham strain was, however, considerably less virulent for rabbits; the number of organisms required for equivalent time of development and severity of skin lesions was 10-fold to 100-fold greater for the Birmingham than for the Nichols strain. The pathology of testicular infection was broadly similar for both strains. PMID:6751463

  10. Structural, bioinformatic, and in vivo analyses of two Treponema pallidum lipoproteins reveal a unique TRAP transporter

    PubMed Central

    Deka, Ranjit K.; Brautigam, Chad A.; Goldberg, Martin; Schuck, Peter; Tomchick, Diana R.; Norgard, Michael V.

    2012-01-01

    Treponema pallidum, the bacterial agent of syphilis, is predicted to encode one tripartite ATP- independent periplasmic transporter (TRAP-T). TRAP-Ts typically employ a periplasmic substrate-binding protein (SBP) to deliver the cognate ligand to the transmembrane symporter. Herein, we demonstrate that the genes encoding the putative TRAP-T components from T. pallidum, tp0957 (the SBP) and tp0958 (the symporter) are in an operon with an uncharacterized third gene, tp0956. We determined the crystal structure of recombinant Tp0956; the protein is trimeric and perforated by a pore. Part of Tp0956 forms an assembly similar to those of “tetratricopeptide repeat” (TPR) motifs. The crystal structure of recombinant Tp0957 was also determined; like the SBPs of other TRAP-Ts, there are two lobes separated by a cleft. In these other SBPs, the cleft binds a negatively charged ligand. However, the cleft of Tp0957 has a strikingly hydrophobic chemical composition, indicating that its ligand may be substantially different and likely hydrophobic. Analytical ultracentrifugation of the recombinant versions of Tp0956 and Tp0957 established that these proteins associate avidly. This unprecedented interaction was confirmed for the native molecules using in vivo cross-linking experiments. Finally, bioinformatic analyses suggested that this transporter exemplifies a new subfamily of TPR-protein associated TRAP transporters (TPATs) that require the action of a TPR-containing accessory protein for the periplasmic transport of a potentially hydrophobic ligand(s). PMID:22306465

  11. Comparison of the locomotor activating effects of bicuculline infusions into the preoptic area and ventral pallidum

    PubMed Central

    Zahm, Daniel S.; Schwartz, Zachary M.; Lavezzi, Heather N.; Yetnikoff, Leora; Parsley, Kenneth P.

    2013-01-01

    Ambulatory locomotion in the rodent is robustly activated by unilateral infusions into the basal forebrain of type A gamma-aminobutyric acid (GABAA) receptor antagonists, such as bicuculline and picrotoxin. The present study was carried out to better localize the neuroanatomical substrate(s) underlying this effect. To accomplish this, differences in total locomotion accumulated during a 20 minute test period following bicuculline versus saline infusions in male Sprague-Dawley rats were calculated, rank ordered and mapped on a diagram of basal forebrain transposed from immunoprocessed sections. The most robust locomotor activation was elicited by bicuculline infusions clustered in rostral parts of the preoptic area. Unilateral infusions of bicuculline into the ventral pallidum produced an unanticipatedly diminutive activation of locomotion, which led us to evaluate bilateral ventral pallidal infusions, and these also produced only a small activation of locomotion, and, interestingly, a non-significant trend toward suppression of rearing. Subjects with bicuculline infused bilaterally into the ventral pallidum also exhibited persistent bouts of abnormal movements. Bicuculline infused unilaterally into other forebrain structures, including the bed nucleus of stria terminalis, caudate-putamen, globus pallidus, sublenticular extended amygdala and sublenticular substantia innominata, did not produce significant locomotor activation. Our data identify the rostral preoptic area as the main substrate for the locomotor activating effects of basal forebrain bicuculline infusions. In contrast, slight activation of locomotion and no effect on rearing accompanied unilateral and bilateral ventral pallidal infusions. Implications of these findings for forebrain processing of reward are discussed. PMID:23423460

  12. Evaluation of Macrolide Resistance and Enhanced Molecular Typing of Treponema pallidum in Patients with Syphilis in Taiwan: a Prospective Multicenter Study

    PubMed Central

    Wu, Hsiu; Chang, Sui-Yuan; Lee, Nan-Yao; Huang, Wen-Chi; Wu, Bing-Ru; Yang, Chia-Jui; Liang, Shiou-Haur; Lee, Chen-Hsiang; Ko, Wen-Chien; Lin, Hsi-Hsun; Chen, Yen-Hsu; Liu, Wen-Chun; Su, Yi-Ching; Hsieh, Chia-Yin; Wu, Pei-Ying

    2012-01-01

    Studies of macrolide resistance mutations and molecular typing using the newly proposed enhanced typing system for Treponema pallidum isolates obtained from HIV-infected patients in the Asia-Pacific region are scarce. Between September 2009 and December 2011, we conducted a survey to detect T. pallidum using a PCR assay using clinical specimens from patients with syphilis at six major designated hospitals for HIV care in Taiwan. The T. pallidum strains were genotyped by following the enhanced molecular typing methodology, which analyzed the number of 60-bp repeats in the acidic repeat protein (arp) gene, T. pallidum repeat (tpr) polymorphism, and the sequence of base pairs 131 to 215 in the tp0548 open reading frame of T. pallidum. Detection of A2058G and A2059G point mutations in the T. pallidum 23S rRNA was performed with the use of restriction fragment length polymorphism (RFLP). During the 2-year study period, 211 clinical specimens were obtained from 136 patients with syphilis. T. pallidum DNA was isolated from 105 (49.8%) of the specimens, with swab specimens obtained from chancres having the highest yield rate (63.2%), followed by plasma (49.4%), serum (35.7%), and cerebrospinal fluid or vitreous fluid (18.2%) specimens. Among the 40 fully typed specimens, 11 subtypes of T. pallidum were identified. Subtype 14f/f (18 isolates) was the most common isolates, followed by 14f/c (3), 14b/c (3), and 14k/f (3). Among the isolates examined for macrolide resistance, none had the A2058G or A2059G mutation. In conclusion, we found that type 14 f/f was the most common T. pallidum strain in this multicenter study on syphilis in Taiwan and that none of the isolates exhibited 23S rRNA mutations causing resistance to macrolides. PMID:22518868

  13. Role of D1 dopamine receptors of the ventral pallidum in inhibitory avoidance learning.

    PubMed

    Péczely, László; Ollmann, Tamás; László, Kristóf; Kovács, Anita; Gálosi, Rita; Szabó, Ádám; Karádi, Zoltán; Lénárd, László

    2014-08-15

    The mesolimbic dopaminergic system (MLDS) originating from the ventral tegmental area has important role in the regulation of motivation, learning and memory. The ventral pallidum (VP), innervated by the MLDS, is involved in the regulation of adaptive behavior, but its exact role is not known in inhibitory avoidance learning. The VP contains both D1 and D2 dopamine receptors, but the density of the former subtype is more excessive. Therefore, in our present experiments, the role of D1 dopamine receptors of the VP in one trial step-through inhibitory avoidance paradigm was investigated. In the conditioning trial, animals were shocked 3 times with 0.5 mA current for 1s, and subsequently were microinjected bilaterally with D1 dopamine receptor agonist SKF38393 into the VP in three doses (0.1 ?g, 1.0 ?g or 5.0 ?g in 0.4 ?l saline). To clarify whether the agonist effect was specific, we also applied the D1 dopamine receptor antagonist SCH23390 (5.0 ?g in 0.4 ?l saline) 15 min prior the agonist treatment. The D1 dopamine receptor agonist, in a dose-dependent manner, significantly increased the step-through latency during the test trials: retention was significant relative to the controls even after 2 weeks of conditioning. The D1 dopamine receptor antagonist SCH23390 pretreatment eliminated SKF38393 effects in the ventral pallidum. Our results show that D1 dopamine receptor mediated mechanisms in the VP facilitate learning and memory in inhibitory avoidance paradigm and this facilitation is specific because it can be eliminated by D1 dopamine receptor antagonist. PMID:24815313

  14. Clovamide-rich extract from Trifolium pallidum reduces oxidative stress-induced damage to blood platelets and plasma

    Microsoft Academic Search

    Joanna Kolodziejczyk; Beata Olas; Barbara Wachowicz; Barbara Szajwaj; Anna Stochmal; Wieslaw Oleszek

    Numerous plants (including clovers) have been widely used in folk medicine for the treatment of different disorders. This\\u000a in vitro study was designed to examine the antioxidative effects of the clovamide-rich fraction, obtained from aerial parts\\u000a of Trifolium pallidum, in the protection of blood platelets and plasma against the nitrative and oxidative damage, caused by peroxynitrite (ONOO?). Carbonyl groups and

  15. Excessive disgust caused by brain lesions or temporary inactivations: Mapping hotspots of nucleus accumbens and ventral pallidum

    PubMed Central

    Ho, Chao-Yi; Berridge, Kent C.

    2014-01-01

    Disgust is a prototypical type of negative affect. In animal models of excessive disgust, only a few brain sites are known in which localized dysfunction (lesions or neural inactivations) can induce intense ‘disgust reactions’ (e.g., gapes) to a normally pleasant sensation such as sweetness. Here we aimed to map forebrain candidates more precisely to identify where either local neuronal damage (excitotoxin lesions) or local pharmacological inactivation (muscimol-baclofen microinjections) caused rats to emit excessive sensory disgust reactions to sucrose. Our study compared subregions of nucleus accumbens shell, ventral pallidum, lateral hypothalamus and adjacent extended amygdala. Results indicated the posterior half of ventral pallidum to be the only forebrain site where intense sensory disgust gapes to sucrose were induced by both lesions and temporary inactivations (this site was previously identified as a hedonic hotspot for enhancements of sweetness ‘liking’). By comparison, for the nucleus accumbens, temporary GABA inactivations in the caudal half of the medial shell also generated sensory disgust but lesions never did at any site. Further, even inactivations failed to induce disgust in the rostral half of accumbens shell (which also contains a hedonic hotspot). In other structures, neither lesions nor inactivations induced disgust as long as the posterior ventral pallidum remained spared. We conclude that the posterior ventral pallidum is an especially crucial hotspot for producing excessive sensory disgust by local pharmacological/lesion dysfunction. By comparison, the nucleus accumbens appears to segregate sites for pharmacological disgust induction and hedonic enhancement into separate posterior versus rostral halves of medial shell. PMID:25229197

  16. The time-dependent clearance of virulent Treponema pallidum in susceptible and resistant strains of guinea pigs is significantly different.

    PubMed

    Wicher, V; Wicher, K; Abbruscato, F; Auger, I; Rudofsky, U

    1999-04-01

    The kinetics of clearance of Treponema pallidum spp. pallidum Nichols from skin and testes of susceptible C4-deficient (C4D) and -resistant Albany (Alb) strains of guinea pigs (gps) was evaluated using the polymerase chain reaction (PCR) and the rabbit infectivity test (RIT). For each strain there were two groups of animals, one infected with virulent T. pallidum (TP) and one control injected with heat-killed treponemes (HKTP). The kinetic studies and their statistical analysis showed that in the C4D strain the microbial clearance in both tissues was significantly slower (p < 0.005) and still incomplete at 3 months after infection. In the Alb strain the clearance was faster and apparently completed within a month. A greater permissiveness in bacterial growth in C4D compared to Alb appears to be one critical factor determining the different rate of local elimination after primary infection. In both strains there was some correlation between the severity and duration of cutaneous lesions and the local persistence of viable organisms. This correlation was not observed in testes. These studies suggest a genetic basis for the strain-specific susceptibility and resistance phenotypes in the pathogenesis of syphilis. PMID:10219257

  17. Antibody Purification AntibodyPurification

    E-print Network

    Lebendiker, Mario

    Antibody Purification AntibodyPurification 24 Overview Antibodies are proteins; therefore, methods forms of general protein purification methods (see the Protein Purification section of the PierceG. Protein L is a third protein of bacterial origin that has been developed for use in affinity purification

  18. Trifolium pallidum and Trifolium scabrum extracts in the protection of human plasma components.

    PubMed

    Kolodziejczyk-Czepas, Joanna; Olas, Beata; Malinowska, Joanna; Wachowicz, Barbara; Moniuszko-Szajwaj, Barbara; Kowalska, Iwona; Oleszek, Wieslaw; Stochmal, Anna

    2013-02-01

    Clovers (genus: Trifolium) have been used in traditional medicine by many cultures, but the biological activity of the most of these plants still remains unknown. The aim of our in vitro study was to assess the antioxidative action of phenolic extracts from aerial parts of Trifolium scabrum and Trifolium pallidum in human blood plasma, exposed to oxidative stress. In the present study we also demonstrate, for the first time the effects of the tested extracts on coagulative properties and fibrinolytic activity of blood plasma. The protective properties of the examined extracts (0.5-50 ?g/ml) against peroxynitrite-induced oxidative stress were estimated by the measurements of 3-nitrotyrosine, thiol groups and the thiobarbituric acid-reactive substances levels. The extracts considerably prevented the oxidative and nitrative damage to plasma proteins. Even the lowest doses of the Trifolium extracts (0.5 ?g/ml) were able to markedly reduce 3-nitrotyrosine formation (by about 50%) and to increase the level of -SH groups (by about 30%), in comparison to the plasma exposed to ONOO(-) in the absence of the extracts. The protective action of all the used concentrations of the Trifolium extracts in the prevention of lipid peroxidation was also found. The tested extracts influenced neither the coagulative properties nor fibrinolytic activity of plasma. Moreover, the extracts were able to significantly reduce the inhibitory effect of ONOO(-) on fibrinolytic activity of plasma (assessed with the use of a chromogenic substrate for plasmin). PMID:23335023

  19. Positive reinforcing effect of neurotensin microinjection into the ventral pallidum in conditioned place preference test.

    PubMed

    Ollmann, Tamás; Péczely, László; László, Kristóf; Kovács, Anita; Gálosi, Rita; Berente, Eszter; Karádi, Zoltán; Lénárd, László

    2015-02-01

    The ventral pallidum (VP) is innervated by the mesolimbic dopaminergic system and it has a key role in motivation, reward, and memory processes. Neurotensin (NT) is an important neuromodulator which has been shown to modulate reinforcement in the ventral tegmental area, in the ventral mesencephalic region and in the central nucleus of amygdala. Neurotensin receptor 1 (NTR1) has already been detected in the VP in abundance, but its role in rewarding and reinforcing processes is not fully understood yet. In our present experiments, the effects of NT on positive reinforcement were investigated in the VP. In conditioned place preference (CPP) test, male Wistar rats were microinjected bilaterally with 100 ng or 250 ng NT in the volume of 0.4 ?l. In other groups of animals, 35 ng NTR1 antagonist SR 48692 was applied by itself, or microinjected 15 min before 100 ng NT treatment. One hundred ng dose of NT induced CPP, whereas animals injected with 250 ng NT did not exhibit significant differences from the vehicle group. Antagonist pretreatment inhibited the effect of NT, while the antagonist applied by itself had no effect. Our results show that NT injected into the VP is involved in positive reinforcement. This effect is specific to NTR1 receptors because the development of CPP can be prevented by specific antagonist. PMID:25447302

  20. Reduction of Nonspecific Background Staining in the Fluorescent Treponemal Antibody-Absorption Test

    PubMed Central

    Roberts, Merritt E.; Miller, James N.; Binnings, Gerald F.

    1968-01-01

    The nonspecific background fluorescence which occurs with the fluorescent treponemal antibody-absorption test for syphilis was found to result from a reaction between serum-treated Treponema pallidum organisms and the conjugated antihuman ?-globulin. It was also shown that ?-lipoprotein and albumin were the important contributing factors in human serum. Various dilutions of 2.5% trypsin in phosphate-buffered saline specifically reduced background fluorescence under proper test conditions. By employing a trypsin digestion method, a semiautomated procedure utilizing a visual readout has been postulated as feasible. PMID:4177869

  1. Cocaine dysregulates opioid gating of GABA neurotransmission in the ventral pallidum.

    PubMed

    Kupchik, Yonatan M; Scofield, Michael D; Rice, Kenner C; Cheng, Kejun; Roques, Bernard P; Kalivas, Peter W

    2014-01-15

    The ventral pallidum (VP) is a target of dense nucleus accumbens projections. Many of these projections coexpress GABA and the neuropeptide enkephalin, a ? and ? opioid receptor (MOR) ligand. Of these two, the MOR in the VP is known to be involved in reward-related behaviors, such as hedonic responses to palatable food, alcohol intake, and reinstatement of cocaine seeking. Stimulating MORs in the VP decreases extracellular GABA, indicating that the effects of MORs in the VP on cocaine seeking are via modulating GABA neurotransmission. Here, we use whole-cell patch-clamp on a rat model of withdrawal from cocaine self-administration to test the hypothesis that MORs presynaptically regulate GABA transmission in the VP and that cocaine withdrawal changes the interaction between MORs and GABA. We found that in cocaine-extinguished rats pharmacological activation of MORs no longer presynaptically inhibited GABA release, whereas blocking the MORs disinhibited GABA release. Moreover, MOR-dependent long-term depression of GABA neurotransmission in the VP was lost in cocaine-extinguished rats. Last, GABA neurotransmission was found to be tonically suppressed in cocaine-extinguished rats. These substantial synaptic changes indicated that cocaine was increasing tone on MOR receptors. Accordingly, increasing endogenous tone by blocking the enzymatic degradation of enkephalin inhibited GABA neurotransmission in yoked saline rats but not in cocaine-extinguished rats. In conclusion, our results indicate that following withdrawal from cocaine self-administration enkephalin levels in the VP are elevated and the opioid modulation of GABA neurotransmission is impaired. This may contribute to the difficulties withdrawn addicts experience when trying to resist relapse. PMID:24431463

  2. Differential roles of ventral pallidum subregions during cocaine self-administration behaviors

    PubMed Central

    Root, David H.; Ma, Sisi; Barker, David J.; Megehee, Laura; Striano, Brendan M.; Ralston, Carla M.; Fabbricatore, Anthony T.; West, Mark O.

    2012-01-01

    The ventral pallidum (VP) is necessary for drug-seeking behavior. VP contains ventromedial (VPvm) and dorsolateral (VPdl) subregions which receive projections from the nucleus accumbens shell and core, respectively. To date, no study has investigated the behavioral functions of the VPdl and VPvm subregions. To address this issue, we investigated whether changes in firing rate (FR) differed between VP subregions during four events: approaching toward, responding on, or retreating away from a cocaine-reinforced operandum, and a cocaine-associated cue. Baseline FR and waveform characteristics did not differ between subregions. VPdl neurons exhibited a greater change in FR compared to VPvm neurons during approaches toward, as well as responses on, the cocaine-reinforced operandum. VPdl neurons were more likely to exhibit a similar change in FR (direction and magnitude) during approach and response than VPvm neurons. In contrast, VPvm firing patterns were heterogeneous, changing FRs during approach or response alone, or both. VP neurons did not discriminate cued behaviors from uncued behaviors. No differences were found between subregions during the retreat and no VP neurons exhibited patterned changes in FR in response to the cocaine-associated cue. The stronger, sustained FR changes of VPdl neurons during approach and response may implicate VPdl in the processing of drug-seeking and drug-taking behavior via projections to subthalamic nucleus and substantia nigra pars reticulata. In contrast, heterogeneous firing patterns of VPvm neurons may implicate VPvm in facilitating mesocortical structures with information related to the sequence of behaviors predicting cocaine self-infusions via projections to mediodorsal thalamus and ventral tegmental area. PMID:22806483

  3. Thyroid Antibodies

    MedlinePLUS

    ... autoimmune disease . A low level of thyroid hormones ( hypothyroidism ) can cause symptoms, such as: Weight gain Fatigue ... drug therapy that has associated risks of developing hypothyroidism when thyroid peroxidase antibodies are present, such as ...

  4. Retention of motility of Treponema pallidum (Nichols virulent strain) in an anaerobic cell culture system and in a cell-free system.

    PubMed Central

    Sandok, P L; Jenkin, H M; Graves, S R; Knight, S T

    1976-01-01

    Optimum parameters for retention of motility of Treponema pallidum (Nichols virulent strain) were found by anaerobic co-incubation of the treponeme with rat glial cells and anaerobic incubation in spent medium obtained from glial cells originally grown aerobically. PMID:767359

  5. Ventral Pallidum Roles in Reward and Motivation Kyle S. Smith1,*, Amy J. Tindell2, J. Wayne Aldridge2,3, and Kent C. Berridge2

    E-print Network

    research interest as a mechanism of reward and incentive motivation. As a major output for limbic signals mechanism of reward in the brain. We review data indicating that 1) an intact ventral pallidum is necessary and motivation signals via phasic bursts of excitation to incentive and hedonic stimuli. We conclude

  6. Evaluation of Two Immunoblot Assays and a Western Blot Assay for the Detection of Antisyphilis Immunoglobulin G Antibodies ?

    PubMed Central

    Welch, Ryan J.; Litwin, Christine M.

    2010-01-01

    In the present study, two immunoglobulin G (IgG) immunoblot assays and one IgG Western blot assay were compared to the rapid plasma reagin test (RPR), the fluorescent treponemal antibody absorption test (FTA-ABS), and the Treponema pallidum particle agglutination assay (TP-PA). The agreement levels of the Viramed, Virotech, and MarDx assays were 97.0%, 96.4%, and 99.4%, and the agreements of samples inconclusive by FTA-ABS and resolved by TP-PA were 91.7%, 83.3%, and 69.4%, respectively. PMID:19940043

  7. Bispecific antibodies.

    PubMed

    Kontermann, Roland E; Brinkmann, Ulrich

    2015-07-01

    Bispecific antibodies (bsAbs) combine specificities of two antibodies and simultaneously address different antigens or epitopes. BsAbs with 'two-target' functionality can interfere with multiple surface receptors or ligands associated, for example with cancer, proliferation or inflammatory processes. BsAbs can also place targets into close proximity, either to support protein complex formation on one cell, or to trigger contacts between cells. Examples of 'forced-connection' functionalities are bsAbs that support protein complexation in the clotting cascade, or tumor-targeted immune cell recruiters and/or activators. Following years of research and development (R&D), the first bsAb was approved in 2009. Another bsAb entered the market in December 2014 and several more are in clinical trials. Here, we describe the potentials of bsAbs to become the next wave of antibody-based therapies, focusing on molecules in clinical development. PMID:25728220

  8. A multicenter prospective trial to asses a new real-time polymerase chain reaction for detection of Treponema pallidum, herpes simplex-1/2 and Haemophilus ducreyi in genital, anal and oropharyngeal ulcers.

    PubMed

    Glatz, M; Juricevic, N; Altwegg, M; Bruisten, S; Komericki, P; Lautenschlager, S; Weber, R; Bosshard, P P

    2014-12-01

    Treponema pallidum, herpes simplex virus types 1 or 2 (HSV-1/2) and Haemophilus ducreyi are sexually transmitted pathogens that can cause genital, anal and oropharyngeal ulcers. Laboratory evaluation of these pathogens in ulcers requires different types of specimens and tests, increasing the risk of improper specimen handling and time lapse until analysis. We sought to develop a new real-time PCR (TP-HD-HSV1/2 PCR) to facilitate the detection of T. pallidum, HSV-1/2 and H. ducreyi in ulcers. The TP-HD-HSV1/2 PCR was tested (i) in a retrospective study on 193 specimens of various clinical origin and (ii) in a prospective study on 36 patients with genital, anal or oropharyngeal ulcers (ClinicalTrials.gov # NCT01688258). The results of the TP-HD-HSV1/2 PCR were compared with standard diagnostic methods (T. pallidum: serology, dark field microscopy; HSV-1/2: PCR; H. ducreyi: cultivation). Sensitivity and specificity of the TP-HD-HSV1/2 PCR for T. pallidum were both 100%, for HSV-1 100% and 98%, and for HSV-2 100% and 98%, respectively. T. pallidum and HSV-1/2 were detected in 53% and 22% of patients in the prospective study; H. ducreyi was not detected. In the prospective study, 5/19 (26%) specimens were true positive for T. pallidum in the TP-HD-HSV1/2 PCR but non-reactive in the VDRL. The TP-HD-HSV1/2 PCR is sensitive and specific for the detection of T. pallidum and HSV-1/2 in routine clinical practice and it appears superior to serology in early T. pallidum infections. PMID:24909546

  9. Multicentre surveillance of prevalence of the 23S rRNA A2058G and A2059G point mutations and molecular subtypes of Treponema pallidum in Taiwan, 2009-2013.

    PubMed

    Wu, B-R; Yang, C-J; Tsai, M-S; Lee, K-Y; Lee, N-Y; Huang, W-C; Wu, H; Lee, C-H; Chen, T-C; Ko, W-C; Lin, H-H; Lu, P-L; Chen, Y-H; Liu, W-C; Yang, S-P; Wu, P-Y; Su, Y-C; Hung, C-C; Chang, S-Y

    2014-08-01

    Resistance mutations A2058G and A2059G, within the 23S rRNA gene of Treponema pallidum, have been reported to cause treatment failures in patients receiving azithromycin for syphilis. Genotyping of T. pallidum strains sequentially isolated from patients with recurrent syphilis is rarely performed. From September 2009 to August 2013, we collected 658 clinical specimens from 375 patients who presented with syphilis for genotyping to examine the number of 60-bp repeats in the acidic repeat protein (arp) gene, T. pallidum repeat (tpr) polymorphism, and tp0548 gene, and to detect A2058G and A2059G point mutations by restriction fragment length polymorphism. Treponemal DNA was identified in 45.2% (n = 298) of the specimens that were collected from 216 (57.6%) patients; 268 (40.7%) specimens tested positive for the 23S rRNA gene, and were examined for macrolide resistance. Two isolates (0.7%) harboured the A2058G mutation, and no A2059G mutation was identified. A total of 14 strains of T. pallidum were identified, with 14f/f (57.5%) and 14b/c (10.0%) being the two predominant strains. Forty patients who presented with recurrent episodes of syphilis had T. pallidum DNA identified from the initial and subsequent episodes, with five cases showing strain discrepancies. One patient had two strains identified from different clinical specimens collected in the same episode. Our findings show that 14f/f is the most common T. pallidum strain in Taiwan, where the prevalence of T. pallidum strains that show A2058G or A2059G mutation remains low. Different genotypes of T. pallidum can be identified in patients with recurrent episodes of syphilis. PMID:24438059

  10. Antibody production

    Microsoft Academic Search

    John R. Birch; Andrew J. Racher

    2006-01-01

    The clinical and commercial success of monoclonal antibodies has led to the need for very large-scale production in mammalian cell culture. This has resulted in rapid expansion of global manufacturing capacity [1], an increase in size of reactors (up to 20,000 L) and a greatly increased effort to improve process efficiency with concomitant manufacturing cost reduction. This has been particularly successful

  11. Evidence for Posttranslational Protein Flavinylation in the Syphilis Spirochete Treponema pallidum: Structural and Biochemical Insights from the Catalytic Core of a Periplasmic Flavin-Trafficking Protein

    PubMed Central

    Deka, Ranjit K.; Brautigam, Chad A.; Liu, Wei Z.

    2015-01-01

    ABSTRACT The syphilis spirochete Treponema pallidum is an important human pathogen but a highly enigmatic bacterium that cannot be cultivated in vitro. T. pallidum lacks many biosynthetic pathways and therefore has evolved the capability to exploit host-derived metabolites via its periplasmic lipoprotein repertoire. We recently reported a flavin-trafficking protein in T. pallidum (Ftp_Tp; TP0796) as the first bacterial metal-dependent flavin adenine dinucleotide (FAD) pyrophosphatase that hydrolyzes FAD into AMP and flavin mononucleotide (FMN) in the spirochete’s periplasm. However, orthologs of Ftp_Tp from other bacteria appear to lack this hydrolytic activity; rather, they bind and flavinylate subunits of a cytoplasmic membrane redox system (Nqr/Rnf). To further explore this dichotomy, biochemical analyses, protein crystallography, and structure-based mutagenesis were used to show that a single amino acid change (N55Y) in Ftp_Tp converts it from an Mg2+-dependent FAD pyrophosphatase to an FAD-binding protein. We also demonstrated that Ftp_Tp has a second enzymatic activity (Mg2+-FMN transferase); it flavinylates protein(s) covalently with FMN on a threonine side chain of an appropriate sequence motif using FAD as the substrate. Moreover, mutation of a metal-binding residue (D284A) eliminates Ftp_Tp’s dual activities, thereby underscoring the role of Mg2+ in the enzyme-catalyzed reactions. The posttranslational flavinylation activity that can target a periplasmic lipoprotein (TP0171) has not previously been described. The observed activities reveal the catalytic flexibility of a treponemal protein to perform multiple functions. Together, these findings imply mechanisms by which a dynamic pool of flavin cofactor is maintained and how flavoproteins are generated by Ftp_Tp locally in the T. pallidum periplasm. PMID:25944861

  12. Syphilis-causing strains belong to separate SS14-like or Nichols-like groups as defined by multilocus analysis of 19 Treponema pallidum strains.

    PubMed

    Nechvátal, Lukáš; P?trošová, Helena; Grillová, Linda; Pospíšilová, Petra; Mikalová, Lenka; Strnadel, Radim; Kuklová, Ivana; Kojanová, Martina; Kreidlová, Miluše; Va?ousová, Daniela; Procházka, P?emysl; Zákoucká, Hana; Krch?áková, Alena; Smajs, David

    2014-07-01

    Treponema pallidum strains are closely related at the genome level but cause distinct diseases. Subspecies pallidum (TPA) is the causative agent of syphilis, subspecies pertenue (TPE) causes yaws while subspecies endemicum (TEN) causes bejel (endemic syphilis). Compared to the majority of treponemal genomic regions, several chromosomal loci were found to be more diverse. To assess genetic variability in diverse genomic positions, we have selected (based on published genomic data) and sequenced five variable loci, TP0304, TP0346, TP0488, TP0515 and TP0558, in 19 reference Treponema pallidum strains including all T. pallidum subspecies (TPA, TPE and TEN). Results of this multilocus analysis divided syphilitic isolates into two groups: SS14-like and Nichols-like. The SS14-like group is comprised of SS14, Grady, Mexico A and Philadelphia 1 strains. The Nichols-like group consisted of strains Nichols, Bal 73-1, DAL-1, MN-3, Philadelphia 2, Haiti B and Madras. The TP0558 locus was selected for further studies because it clearly distinguished between the SS14- and Nichols-like groups and because the phylogenetic tree derived from the TP0558 locus showed the same clustering pattern as the tree constructed from whole genome sequences. In addition, TP0558 was shown as the only tested locus that evolved under negative selection within TPA strains. Sequencing of a short fragment (573bp) of the TP0558 locus in a set of 25 clinical isolates from 22 patients collected in the Czech Republic during 2012-2013 revealed that clinical isolates follow the SS14- and Nichols-like distribution. PMID:24841252

  13. Antibody Engineering and Therapeutics

    PubMed Central

    Almagro, Juan Carlos; Gilliland, Gary L; Breden, Felix; Scott, Jamie K; Sok, Devin; Pauthner, Matthias; Reichert, Janice M; Helguera, Gustavo; Andrabi, Raiees; Mabry, Robert; Bléry, Mathieu; Voss, James E; Laurén, Juha; Abuqayyas, Lubna; Barghorn, Stefan; Ben-Jacob, Eshel; Crowe, James E; Huston, James S; Johnston, Stephen Albert; Krauland, Eric; Lund-Johansen, Fridtjof; Marasco, Wayne A; Parren, Paul WHI; Xu, Kai Y

    2014-01-01

    The 24th Antibody Engineering & Therapeutics meeting brought together a broad range of participants who were updated on the latest advances in antibody research and development. Organized by IBC Life Sciences, the gathering is the annual meeting of The Antibody Society, which serves as the scientific sponsor. Preconference workshops on 3D modeling and delineation of clonal lineages were featured, and the conference included sessions on a wide variety of topics relevant to researchers, including systems biology; antibody deep sequencing and repertoires; the effects of antibody gene variation and usage on antibody response; directed evolution; knowledge-based design; antibodies in a complex environment; polyreactive antibodies and polyspecificity; the interface between antibody therapy and cellular immunity in cancer; antibodies in cardiometabolic medicine; antibody pharmacokinetics, distribution and off-target toxicity; optimizing antibody formats for immunotherapy; polyclonals, oligoclonals and bispecifics; antibody discovery platforms; and antibody-drug conjugates. PMID:24589717

  14. Macrophages in immunity to syphilis: suppressive effect of concurrent infection with Mycobacterium bovis BCG on the development of syphilitic lesions and growth of Treponema pallidum in tuberculin-positive rabbits.

    PubMed Central

    Hardy, P H; Graham, D J; Nell, E E; Dannenberg, A M

    1979-01-01

    Paired groups of male rabbits were challenged with Treponema pallidum and Mycobacterium bovis BCG. One group had been sensitized to BCG by inoculation 3 weeks before challenge. All animals were challenged intradermally at multiple sites with T. pallidum alone, BCG alone, and both organisms into the same sites. The resulting lesions were followed clinically and histologically. BCG lesions enlarged more rapidly in sensitized rabbits, but they were otherwise no different from those in the controls. T. pallidum lesions enlarged and regressed simultaneously in both groups, but in the BCG-sensitized animals they became twice as large as those in the unsensitized rabbits. Mixed BCG-T. pallidum lesions showed the greatest differences in the two groups of animals. Like the pure BCG lesions, they enlarged more rapidly in the sensitized rabbits but began to recede after 1 week. The corresponding lesions in the controls enlarged more slowly and reached their maximum size after 3 weeks when the receding lesions in the sensitized animals were much smaller. The most marked histological-histochemical difference between the two groups of animals was in the number and activation of macrophages. These cells were more numerous in the mixed lesions of BCG-sensitized animals than in similar lesions of the controls and more activated as determined by beta-galactosidase staining. Although sparsely distributed, activated macrophages were more numerous in the pur T. pallidum lesions of sensitized animals than in those of control animals. Silver-stained sections revealed fewer treponemes in mixed lesions of sensitized animals than in the mixed lesions of control animals. Quantitation of treponemes in pure T. pallidum versus mixed lesions was determined in two groups of rabbits challenged intratesticularly. The total number of treponemes per testis in the mixed lesions of BCG-sensitized rabbits was significantly less than the number in the mixed lesions of control animals, and also less than the number in pure T. pallidum lesions of both groups of animals. Images PMID:397934

  15. Macrophages in immunity to syphilis: suppressive effect of concurrent infection with Mycobacterium bovis BCG on the development of syphilitic lesions and growth of Treponema pallidum in tuberculin-positive rabbits.

    PubMed

    Hardy, P H; Graham, D J; Nell, E E; Dannenberg, A M

    1979-11-01

    Paired groups of male rabbits were challenged with Treponema pallidum and Mycobacterium bovis BCG. One group had been sensitized to BCG by inoculation 3 weeks before challenge. All animals were challenged intradermally at multiple sites with T. pallidum alone, BCG alone, and both organisms into the same sites. The resulting lesions were followed clinically and histologically. BCG lesions enlarged more rapidly in sensitized rabbits, but they were otherwise no different from those in the controls. T. pallidum lesions enlarged and regressed simultaneously in both groups, but in the BCG-sensitized animals they became twice as large as those in the unsensitized rabbits. Mixed BCG-T. pallidum lesions showed the greatest differences in the two groups of animals. Like the pure BCG lesions, they enlarged more rapidly in the sensitized rabbits but began to recede after 1 week. The corresponding lesions in the controls enlarged more slowly and reached their maximum size after 3 weeks when the receding lesions in the sensitized animals were much smaller. The most marked histological-histochemical difference between the two groups of animals was in the number and activation of macrophages. These cells were more numerous in the mixed lesions of BCG-sensitized animals than in similar lesions of the controls and more activated as determined by beta-galactosidase staining. Although sparsely distributed, activated macrophages were more numerous in the pur T. pallidum lesions of sensitized animals than in those of control animals. Silver-stained sections revealed fewer treponemes in mixed lesions of sensitized animals than in the mixed lesions of control animals. Quantitation of treponemes in pure T. pallidum versus mixed lesions was determined in two groups of rabbits challenged intratesticularly. The total number of treponemes per testis in the mixed lesions of BCG-sensitized rabbits was significantly less than the number in the mixed lesions of control animals, and also less than the number in pure T. pallidum lesions of both groups of animals. PMID:397934

  16. Antibody production.

    PubMed

    Birch, John R; Racher, Andrew J

    2006-08-01

    The clinical and commercial success of monoclonal antibodies has led to the need for very large-scale production in mammalian cell culture. This has resulted in rapid expansion of global manufacturing capacity [1], an increase in size of reactors (up to 20,000 L) and a greatly increased effort to improve process efficiency with concomitant manufacturing cost reduction. This has been particularly successful in the upstream part of the process where productivity of cell cultures has improved 100 fold in the last 15 years. This success has resulted from improvements in expression technology and from process optimisation, especially the development of fed-batch cultures. In addition to improving process/cost efficiencies, a second key area has been reducing the time taken to develop processes and produce the first material required for clinical testing and proof-of-principle. Cell line creation is often the slowest step in this stage of process development. This article will review the technologies currently used to make monoclonal antibodies with particular emphasis on mammalian cell culture. Likely future trends are also discussed. PMID:16822577

  17. Lateral hypothalamus, nucleus accumbens, and ventral pallidum roles in eating and hunger: interactions between homeostatic and reward circuitry

    PubMed Central

    Castro, Daniel C.; Cole, Shannon L.; Berridge, Kent C.

    2015-01-01

    The study of the neural bases of eating behavior, hunger, and reward has consistently implicated the lateral hypothalamus (LH) and its interactions with mesocorticolimbic circuitry, such as mesolimbic dopamine projections to nucleus accumbens (NAc) and ventral pallidum (VP), in controlling motivation to eat. The NAc and VP play special roles in mediating the hedonic impact (“liking”) and motivational incentive salience (“wanting”) of food rewards, and their interactions with LH help permit regulatory hunger/satiety modulation of food motivation and reward. Here, we review some progress that has been made regarding this circuitry and its functions: the identification of localized anatomical hedonic hotspots within NAc and VP for enhancing hedonic impact; interactions of NAc/VP hedonic hotspots with specific LH signals such as orexin; an anterior-posterior gradient of sites in NAc shell for producing intense appetitive eating vs. intense fearful reactions; and anatomically distributed appetitive functions of dopamine and mu opioid signals in NAc shell and related structures. Such findings help improve our understanding of NAc, VP, and LH interactions in mediating affective and motivation functions, including “liking” and “wanting” for food rewards. PMID:26124708

  18. [Laboratory diagnosis of Treponema pallidum infection in patients with early syphilis and neurosyphilis through a PCR-based test].

    PubMed

    García, Patricia; Grassi, Bruno; Fich, Félix; Salvo, Aurelio; Araya, Luis; Abarzúa, Fernando; Soto, Julia; Poggi, Helena; Lagos, Marcela; Vásquez, Patricia; León, Eugenia P; Pérez, Carlos; Wozniak, Aniela

    2011-08-01

    Syphilis is a sexually transmitted disease caused by Treponema pallidum. The diagnosis is based mainly in clinical presentation and non-specific assays. PCR-based diagnosis has been suggested as an attractive alternative method. The aim of this study was the validation of a PCR-based test for the diagnosis of early syphilis (ES) and neurosyphilis (NS). Clinical samples of mucocutaneous lesions and cerebrospinal fluid (CSF) specimens from patients previously diagnosed for ES and NS respectively using an enlarged gold standard, were tested by PCR. The reaction was done using primers targeting the tpN47 gene. Twenty out of 21 mucocutaneous samples from patients diagnosed with ES were positive by PCR, with a clinical sensitivity of 95%. Four out of 8 CSF samples from patients previously diagnosed with NS were positive by PCR, with a clinical sensitivity of 50%. The clinical specificity for both ES and NS was 100%. The PCR sensitivity and specificity for mucocutaneous samples allowed us to implement this assay in our laboratory for routine diagnosis. Although the sensitivity of the PCR in CSF was low, it may be useful to support clinical diagnosis. PMID:22052394

  19. The Social Amoeba Polysphondylium pallidum Loses Encystation and Sporulation, but Can Still Erect Fruiting Bodies in the Absence of Cellulose

    PubMed Central

    Du, Qingyou; Schaap, Pauline

    2014-01-01

    Amoebas and other freely moving protists differentiate into walled cysts when exposed to stress. As cysts, amoeba pathogens are resistant to biocides, preventing treatment and eradication. Lack of gene modification procedures has left the mechanisms of encystation largely unexplored. Genetically tractable Dictyostelium discoideum amoebas require cellulose synthase for formation of multicellular fructifications with cellulose-rich stalk and spore cells. Amoebas of its distant relative Polysphondylium pallidum (Ppal), can additionally encyst individually in response to stress. Ppal has two cellulose synthase genes, DcsA and DcsB, which we deleted individually and in combination. Dcsa- mutants formed fruiting bodies with normal stalks, but their spore and cyst walls lacked cellulose, which obliterated stress-resistance of spores and rendered cysts entirely non-viable. A dcsa-/dcsb- mutant made no walled spores, stalk cells or cysts, although simple fruiting structures were formed with a droplet of amoeboid cells resting on an sheathed column of decaying cells. DcsB is expressed in prestalk and stalk cells, while DcsA is additionally expressed in spores and cysts. We conclude that cellulose is essential for encystation and that cellulose synthase may be a suitable target for drugs to prevent encystation and render amoeba pathogens susceptible to conventional antibiotics. PMID:25113829

  20. Enhanced Extracellular Glutamate and Dopamine in the Ventral Pallidum of Alcohol-Preferring AA and Alcohol-Avoiding ANA Rats after Morphine

    PubMed Central

    Kemppainen, Heidi; Nurmi, Harri; Raivio, Noora; Kiianmaa, Kalervo

    2015-01-01

    The purpose of the present study was to investigate the role of ventral pallidal opioidergic mechanisms in the control of ethanol intake by studying the effects of acute administration of morphine on the levels of GABA, glutamate, and dopamine in the ventral pallidum. The study was conducted using the alcohol-preferring Alko Alcohol (AA) and alcohol-avoiding Alko Non-Alcohol (ANA) rat lines that have well-documented differences in their voluntary ethanol intake and brain opioidergic systems. Therefore, examination of neurobiological differences between the lines is supposed to help to identify the neuronal mechanisms underlying ethanol intake, since selection pressure is assumed gradually to lead to enrichment of alleles promoting high or low ethanol intake, respectively. The effects of an acute dose of morphine (1 or 10?mg/kg s.c.) on the extracellular levels of GABA and glutamate in the ventral pallidum were monitored with in vivo microdialysis. The concentrations of GABA and glutamate in the dialyzates were determined with a high performance liquid chromatography system using fluorescent detection, while electrochemical detection was used for dopamine. The levels of glutamate in the rats injected with morphine 1?mg/kg were significantly above the levels found in the controls and in the rats receiving morphine 10?mg/kg. Morphine 10?mg/kg also increased the levels of dopamine. Morphine could not, however, modify the levels of GABA. The rat lines did not differ in any of the effects of morphine. The data suggest that the glutamatergic and dopaminergic systems in the ventral pallidum may mediate some effects of morphine. Since there were no differences between the AA and ANA lines, the basic hypothesis underlying the use of the genetic animal model suggests that the effects of morphine detected probably do not underlie the different intake of ethanol by the lines and contribute to the control of ethanol intake in these animals. PMID:25653621

  1. The Tp38 (TpMglB-2) lipoprotein binds glucose in a manner consistent with receptor function in Treponema pallidum.

    PubMed

    Deka, Ranjit K; Goldberg, Martin S; Hagman, Kayla E; Norgard, Michael V

    2004-04-01

    A 38-kDa lipoprotein of Treponema pallidum (Tp38) was predicted to be a periplasmic sugar-binding protein based on its sequence similarity to the glucose/galactose-binding (MglB) protein of Escherichia coli (P. S. Becker, D. R. Akins, J. D. Radolf, and M. V. Norgard, Infect. Immun. 62:1381-1391, 1994). Inasmuch as glucose is believed to be the principal, if not sole, carbon and energy source for T. pallidum and is readily available to the spirochete during its obligate infection of humans, we hypothesized that Tp38 may serve as the organism's requisite glucose receptor. For the present study, a nonacylated recombinant form of Tp38 was coexpressed with GroES and GroEL in E. coli to facilitate the isolation of soluble, properly folded Tp38. The highly sensitive method of intrinsic fluorescence spectroscopy, predicated on the manner in which tryptophan residues reside and move within protein microenvironments, was then used to assess sugar binding to Tp38. The intrinsic fluorescence of Tp38 was essentially unaltered when it was exposed to D-mannose, D-fucose, D-ribose, L-glucose, or L-galactose, but it changed markedly in the presence of D-glucose, and to a lesser extent, D-galactose, indicating binding. The K(d) values for D-glucose and D-galactose binding to Tp38 were 152.2 +/- 20.73 nM and 251.2 +/- 55.25 nM, respectively. Site-directed mutagenesis of Trp-145, a residue postulated to contribute to the sugar-binding pocket in a manner akin to the essential Trp-183 in E. coli MglB, abolished Tp38's conformational change in response to D-glucose. The combined data are consistent with Tp38 serving as a glucose receptor for T. pallidum. These findings potentially have important implications for syphilis pathogenesis, particularly as they may pertain to glucose-mediated chemotactic responses by T. pallidum. PMID:15060032

  2. [Prevalence, risk factors and genetic characterization of human T-cell lymphotropic virus types 1 and 2 in patients infected with human immunodeficiency virus type 1 in the cities of Ribeirăo Preto and Săo Paulo].

    PubMed

    Kleine Neto, Walter; Sanabani, Sabri Saeed; Jamal, Leda Fátima; Sabino, Ester Cerdeira

    2009-01-01

    The aim of this study was to define the prevalence of human T cell lymphotropic virus types 1 and 2 in patients who were positive for human immunodeficiency virus type 1 in the State of Săo Paulo, Brazil. We evaluated 319 individuals infected with HIV type 1 who were attended at specialized clinics in two cities (Ribeirăo Preto and Săo Paulo). The patients were interviewed and tested for antibodies against HTLV types 1 and 2 (Orthoâ HTLV-1/HTLV-2 Ab-Capture enzyme immunoassay). Direct DNA sequencing of polymerase chain reaction products from the tax region of HTLV type 2 and the long terminal repeat region of HTLV types 1 and 2 were performed to differentiate and determine the subtypes. The overall prevalence of anti-HTLV type 1 and 2 antibodies was 7.5% (24/319; 95% CI: 5.2-11.5). HTLV type 1 and 2 infection was associated with a history of injected drug use and with antibodies for hepatitis C virus (p < 0.001), but not with age (p = 0.2), sex (p = 0.9), sexual behavior or serological markers for sexually transmitted diseases (anti-Treponema pallidum, anti-human herpesvirus type 8 or anti-hepatitis B virus antibodies) (p > 0.05). HTLV DNA was detected in 13 out of 24 samples, of which 12 were characterized as HTLV subtype 2c and one as HTLV subtype 1a. Among the 12 HTLV type 2 samples, seven were from injected drug users, thus indicating that this route is an important risk factor for HTLV type 2 transmission among our population infected with HIV type 1. PMID:19684973

  3. The TP0796 lipoprotein of Treponema pallidum is a bimetal-dependent FAD pyrophosphatase with a potential role in flavin homeostasis.

    PubMed

    Deka, Ranjit K; Brautigam, Chad A; Liu, Wei Z; Tomchick, Diana R; Norgard, Michael V

    2013-04-19

    Treponema pallidum, an obligate parasite of humans and the causative agent of syphilis, has evolved the capacity to exploit host-derived metabolites for its survival. Flavin-containing compounds are essential cofactors that are required for metabolic processes in all living organisms, and riboflavin is a direct precursor of the cofactors FMN and FAD. Unlike many pathogenic bacteria, Treponema pallidum cannot synthesize riboflavin; we recently described a flavin-uptake mechanism composed of an ABC-type transporter. However, there is a paucity of information about flavin utilization in bacterial periplasms. Using a discovery-driven approach, we have identified the TP0796 lipoprotein as a previously uncharacterized Mg(2+)-dependent FAD pyrophosphatase within the ApbE superfamily. TP0796 probably plays a central role in flavin turnover by hydrolyzing exogenously acquired FAD, yielding AMP and FMN. Biochemical and structural investigations revealed that the enzyme has a unique bimetal Mg(2+) catalytic center. Furthermore, the pyrophosphatase activity is product-inhibited by AMP, indicating a possible role for this molecule in modulating FMN and FAD levels in the treponemal periplasm. The ApbE superfamily was previously thought to be involved in thiamine biosynthesis, but our characterization of TP0796 prompts a renaming of this superfamily as a periplasmic flavin-trafficking protein (Ftp). TP0796 is the first structurally and biochemically characterized FAD pyrophosphate enzyme in bacteria. This new paradigm for a bacterial flavin utilization pathway may prove to be useful for future inhibitor design. PMID:23447540

  4. THE PRESERVATION OF VIRULENT TREPONEMA PALLIDUM AND TREPONEMA PERTENUE IN THE FROZEN STATE; WITH A NOTE ON THE PRESERVATION OF FILTRABLE VIRUSES

    PubMed Central

    Turner, Thomas B.

    1938-01-01

    1. A simple method for freezing and maintaining tissue specimens in a mixture of solid carbon dioxide and 95 per cent ethyl alcohol at a temperature approximating –78°C. is described. 2. When frozen and maintained at this temperature Treponema pallidum and Treponema pertenue, upon thawing, exhibited normal morphology and motility and their virulence for rabbits was not appreciably altered after periods of at least 1 year. This applied to a number of different strains of each organism. The infectivity of material in which treponemes were scant was maintained as well as of material in which they were abundant. 3. At temperatures of –10°C. and –20°C. syphilis treponemes did not survive as long as 2 months. Death of the organism occurred not at the time of freezing but during the maintenance period. 4. Treponemes did not commonly survive freezing and desiccation, although one lot of dried material which contained T. pallidum was infective for rabbits 1 day after desiccation. 5. The viruses of human influenza, yellow fever, and spontaneous encephalomyelitis of mice when frozen and maintained at –78°C. showed substantially the same titer after 6 months as before freezing. 6. Certain practical applications of the method are suggested. PMID:19870710

  5. Serologic analyses of cottontail rabbits for antibodies to Borrelia burgdorferi.

    PubMed Central

    Magnarelli, L A; Anderson, J F; McAninch, J B

    1990-01-01

    An enzyme-linked immunosorbent assay was developed to detect antibodies to Borrelia burgdorferi in cottontail rabbits captured in Millbrook, N.Y., and New York, N.Y. Five antigenically variable strains of B. burgdorferi were analyzed to determine the variability of serologic test results. In analyses of 79 serum samples, seropositivity ranged from 56% for a strain cultured from kidney tissues of a cottontail rabbit to 68% for a strain isolated from a larva of Ixodes dentatus, a tick that parasitized a cottontail rabbit. There were false-positive results when reference rabbit antisera to B. hermsii and Treponema pallidum were screened against B. burgdorferi. Cross-reactivity with antisera to Leptospira interrogans serovars was less pronounced. Western blot (immunoblot) analyses revealed reactivities of test sera to two or more surface or subsurface proteins of B. burgdorferi with approximate molecular masses of 18, 25 to 27, 34, 36, 41, and 59 kilodaltons. Cottontail rabbits respond immunologically to B. burgdorferi, but the observed variations in serologic test results should not be a limitation in field and laboratory investigations of Lyme borreliosis. PMID:2351732

  6. Detection of Treponema pallidum in tissue: a comparative study of the avidin-biotin-peroxidase complex, indirect immunoperoxidase, FTA-ABS complement techniques and the darkfield method.

    PubMed

    Lee, W S; Lee, M G; Chung, K Y; Lee, J B

    1991-12-01

    With 37 formalin-fixed, paraffin embedded specimens from the lesions of 30 patients with primary, secondary or gastric syphilis, we performed avidin-biotin-peroxidase complex (ABC), indirect immunoperoxidase (IIP) and FTA-ABS complement techniques. Darkfield examination was done in 17 skin lesions. The immunoperoxidase technique, especially the ABC technique, revealed higher reactivity than the FTA-ABS complement technique and darkfield examination in detecting Treponema pallidum in tissues. Furthermore, the ABC technique produced less intense nonspecific background staining than the IIP technique. Histologically, most of the treponemes were located in the upper dermis, epidermis and vessel walls in the order named, and rarely in the lower dermis of the syphilitic skin lesions. PMID:1812655

  7. Proposal of a new halobacterial genus Natrinema gen. nov., with two species Natrinema pellirubrum nom. nov. and Natrinema pallidum nom. nov.

    PubMed

    McGenity, T J; Gemmell, R T; Grant, W D

    1998-10-01

    A phylogenetic analysis of 69 halobacterial 16S rRNA gene sequences has been carried out, integrating data from new isolates, previously described halobacteria and cloned sequences from uncultivated halobacteria. Halobacterium halobium NCIMB 777, Halobacterium trapanicum NCIMB 784 and Halobacterium salinarium NCIMB 786, together with several other strains (strains T5.7, L11 and Halobacterium trapanicum NCIMB 767) constitute a distinct lineage with at least 98.2% sequence similarity. These strains have been incorrectly assigned to the genus Halobacterium. Therefore, based on a variety of taxonomic criteria, it is proposed that Halobacterium salinarium NCIMB 786 is renamed as Natrinema pellirubrum nom. nov., the type species of the new genus Natrinema gen. nov., and that Halobacterium halobium NCIMB 777 and Halobacterium trapanicum NCIMB 784 are renamed as a single species, Natrinema pallidum nom. nov. It was notable that halobacteria closely related to the proposed new genus have been isolated from relatively low-salt environments. PMID:9828420

  8. Therapeutic antibody expression technology.

    PubMed

    Chadd, H E; Chamow, S M

    2001-04-01

    With the technological advances made during the past decade, antibodies now represent an important and growing class of biotherapeutics. With the potential new targets resulting from genomics and with methods now in place to make fully human antibodies, the potential of antibodies as valuable therapeutics in oncology, inflammation and cardiovascular disease can be fully realised. Systems to produce these antibodies as full-length molecules and as fragments include expression in both mammalian and bacterial cells grown in bioreactors and in transgenic organisms. Factors including molecular fidelity and the cost of goods are critical in evaluating expression systems. Mammalian cell culture and transgenic organisms show the greatest promise for the expression of full-length, recombinant human antibodies, and bacterial fermentation seems most favorable for the expression of antibody fragments. PMID:11287236

  9. Biobarcodes: Antibodies and Nanosensors

    NSDL National Science Digital Library

    Nanoscale Informal Science Education Network

    2014-06-04

    In this activity/demo, learners investigate biobarcodes, a nanomedical technology that allows for massively parallel testing that can assist with disease diagnosis. Learners define antibodies and learn how each antibody binds to a unique protein. Learners also discover how biobarcoding uses nanoparticles, antibodies, DNA and magnetism to detect diseases earlier than we could detect before. Learners assemble a jigsaw puzzle that models how biobarcodes work.

  10. Red Blood Cell Antibody Identification

    MedlinePLUS

    ... Antibody ID, RBC; RBC Ab ID Formal name: Red Blood Cell Antibody Identification Related tests: Direct Antiglobulin ... else I should know? How is it used? Red blood cell antibody identification is used as a ...

  11. Circulating Antibodies in Dermatophytosis

    Microsoft Academic Search

    Sarah F. Grappel; F. Blank; C. T. Bishop

    1972-01-01

    Antibodies to dermatophytes (M. audouinii, T. mentagrophytes var. granulosum and var. interdigitale, T. rubrum, and T. tonsurans) were detected in sera of patients with tinea capitis and tinea corporis by charcoal agglutination tests, immunodiffusion analysis, and complement fixation tests. In tinea capitis, these antibodies were not confined to patients with deep-seated, inflammatory infections. The zoophilic variety of T. mentagrophytes induced

  12. Antibodies in Plants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The expression of antibodies in plants has several promising applications that are currently being developed. Plants are being considered for the large scale production of antibodies needed for medical purposes. The benefit of using plants is that they are able to perform post-translational modifi...

  13. Antibodies for biodefense

    PubMed Central

    Froude, Jeffrey W; Stiles, Bradley; Pelat, Thibaut

    2011-01-01

    Potential bioweapons are biological agents (bacteria, viruses and toxins) at risk of intentional dissemination. Biodefense, defined as development of therapeutics and vaccines against these agents, has seen an increase, particularly in the US, following the 2001 anthrax attack. This review focuses on recombinant antibodies and polyclonal antibodies for biodefense that have been accepted for clinical use. These antibodies aim to protect against primary potential bioweapons or category A agents as defined by the Centers for Disease Control and Prevention (Bacillus anthracis, Yersinia pestis, Francisella tularensis, botulinum neurotoxins, smallpox virus and certain others causing viral hemorrhagic fevers) and certain category B agents. Potential for prophylactic use is presented, as well as frequent use of oligoclonal antibodies or synergistic effect with other molecules. Capacities and limitations of antibodies for use in biodefense are discussed, and are generally applicable to the field of infectious diseases. PMID:22123065

  14. [Anti-PCNA antibody].

    PubMed

    Yamanaka, K; Takasaki, Y

    1993-08-01

    Antibodies to the proliferating cell nuclear antigen (PCNA) detected in patients with systemic lupus erythematosus (SLE) were allowed to react with a nuclear antigen expressed predominantly in proliferating cells such as cultured cells and mitogen-transformed cells. The characterization of both structure and function of PCNA has been studied. PCNA has been identified as a protein with a molecular weight of about 33 kD and isoelectric point of 4.8. The expression of PCNA increased in the cell from the late G1 and S phases of the cell cycle immediately preceding DNA synthesis. Recent studies have revealed that the auxiliary protein of DNA polymerase-d is identical to PCNA. Anti-PCNA antibodies can be detected by the methods of immunofluorescence (IF), double immunodiffusion (DID), counterimmunoelectrophoresis (CIE), enzyme-linked immunosorbent assay (ELISA) and immunoblotting (IB). Anti-PCNA antibodies were specifically detected in 2-5% of the patients with SLE by DID. In spite of their low frequency, anti-PCNA antibodies are useful as a clinical marker for SLE. Several reports indicated that patients with PCNA positive SLE showed a high frequency of renal and central nerve system (CNS) involvements and thrombocytopenia. In these patients, the anti-PCNA antibody titer was elevated before the development by proteinuria and the titer of anti-PCNA antibodies was decreased by treatment with corticosteroids. In addition, anti-PCNA antibodies have been known as a useful tool to detect activated lymphocytes and malignant proliferating cell. PMID:8103808

  15. Anti-sulfotyrosine antibodies

    DOEpatents

    Bertozzi, Carolyn R. (Berkeley, CA); Kehoe, John (Saint Davids, PA); Bradbury, Andrew M. (Santa Fe, NM)

    2009-09-15

    The invention provides anti-sulfotyrosine specific antibodies capable of detecting and isolating polypeptides that are tyrosine-sulfated. The sulfotyrosine antibodies and antibody fragments of the invention may be used to discriminate between the non-sulfated and sulfated forms of such proteins, using any number of immunological assays, such ELISAs, immunoblots, Western Blots, immunoprecipitations, and the like. Using a phage-display system, single chain antibodies (scFvs) were generated and screened against tyrosine-sulfated synthetic peptide antigens, resulting in the isolation of scFvs that specifically recognize sulfotyrosine-containing peptides and/or demonstrate sulfotyrosine-specific binding in tyrosine sulfated proteins. The VH and VL genes from one such sulfotyrosine-specific scFv were employed to generate a full length, sulfotyrosine-specific immunoglobulin.

  16. Antithyroid microsomal antibody

    MedlinePLUS

    ... to confirm the cause of thyroid problems, including Hashimoto thyroiditis . The test is also used to find ... positive test may be due to: Granulomatous thyroiditis Hashimoto thyroiditis High levels of these antibodies have also ...

  17. Mining human antibody repertoires

    PubMed Central

    2010-01-01

    Human monoclonal antibodies (mAbs) have become drugs of choice for the management of an increasing number of human diseases. Human antibody repertoires provide a rich source for human mAbs. Here we review the characteristics of natural and non-natural human antibody repertoires and their mining with non-combinatorial and combinatorial strategies. In particular, we discuss the selection of human mAbs from naďve, immune, transgenic and synthetic human antibody repertoires using methods based on hybridoma technology, clonal expansion of peripheral B cells, single-cell PCR, phage display, yeast display and mammalian cell display. Our reliance on different strategies is shifting as we gain experience and refine methods to the efficient generation of human mAbs with superior pharmacokinetic and pharmacodynamic properties. PMID:20505349

  18. Chryseobacterium ureilyticum sp. nov., Chryseobacterium gambrini sp. nov., Chryseobacterium pallidum sp. nov. and Chryseobacterium molle sp. nov., isolated from beer-bottling plants.

    PubMed

    Herzog, Peter; Winkler, Ilka; Wolking, Dorothee; Kämpfer, Peter; Lipski, André

    2008-01-01

    Four Gram-negative, rod-shaped, non-spore-forming and non-motile bacterial strains were isolated from surfaces and biofilms associated with beer-bottling plants. Based on their 16S rRNA gene sequences these isolates were allocated to the genus Chryseobacterium. The sequence similarities of the isolates to the next most closely related type strains of this genus ranged from 96.4 to 98.3%. The presence of menaquinone MK-6 and predominant fatty acids 15:0 iso, 17:1 iso cis9, 15:0 iso 2-OH and 17:0 iso 3-OH supported the affiliation of these strains to the genus. The results of DNA-DNA hybridization, biochemical tests and chemotaxonomic properties allowed genotypic and phenotypic differentiation of the strains from the next most closely related Chryseobacterium species with validly published names. Therefore, the isolates represent four novel species for which the names Chryseobacterium ureilyticum (type strain F-Fue-04IIIaaaa(T)=DSM 18017(T)=CCUG 52546(T)), Chryseobacterium gambrini (type strain 5-1St1a(T)=DSM 18014(T)=CCUG 52549(T)), Chryseobacterium pallidum (type strain 26-3St2b(T)=DSM 18015(T)=CCUG 52548(T)) and Chryseobacterium molle (type strain DW3(T)=DSM 18016(T)=CCUG 52547(T)) are proposed. PMID:18175677

  19. Glycoproteomic Analysis of Antibodies*

    PubMed Central

    Zauner, Gerhild; Selman, Maurice H. J.; Bondt, Albert; Rombouts, Yoann; Blank, Dennis; Deelder, André M.; Wuhrer, Manfred

    2013-01-01

    Antibody glycosylation has been shown to change with various processes. This review presents mass spectrometric approaches for antibody glycosylation analysis at the level of released glycans, glycopeptides, and intact protein. With regard to IgG fragment crystallizable glycosylation, mass spectrometry has shown its potential for subclass-specific, high-throughput analysis. In contrast, because of the vast heterogeneity of peptide moieties, fragment antigen binding glycosylation analysis of polyclonal IgG relies entirely on glycan release. Next to IgG, IgA has gained some attention, and studies of its O- and N-glycosylation have revealed disease-associated glycosylation changes. Glycoproteomic analyses of IgM and IgE are lagging behind but should complete our picture of glycosylation's influence on antibody function. PMID:23325769

  20. Selection of recombinant antibodies from antibody gene libraries.

    PubMed

    Hust, Michael; Frenzel, André; Schirrmann, Thomas; Dübel, Stefan

    2014-01-01

    Antibodies are indispensable detection reagents for research and diagnostics and represent the biggest class of biological therapeutics on the market. In vitro antibody selection systems offer many advantages over animal-based technologies because the whole selection process is independent of the in vivo immune response. In the last two decades antibody phage display has evolved to the most robust and widely used method and has already yielded thousands of antibodies. The selection of binders by phage display is also referred to as "panning" and based on the specific molecular interaction of antibody phage with an immobilized antigen thus allowing the enrichment and isolation of antigen-specific monoclonal binders from very large antibody gene libraries. Here, we give detailed protocols for the selection of recombinant antibody fragments from antibody gene libraries in microtiter plates. PMID:24233787

  1. Human Germline Antibody Gene Segments Encode Polyspecific Antibodies

    PubMed Central

    Willis, Jordan R.; Briney, Bryan S.; DeLuca, Samuel L.; Crowe, James E.; Meiler, Jens

    2013-01-01

    Structural flexibility in germline gene-encoded antibodies allows promiscuous binding to diverse antigens. The binding affinity and specificity for a particular epitope typically increase as antibody genes acquire somatic mutations in antigen-stimulated B cells. In this work, we investigated whether germline gene-encoded antibodies are optimal for polyspecificity by determining the basis for recognition of diverse antigens by antibodies encoded by three VH gene segments. Panels of somatically mutated antibodies encoded by a common VH gene, but each binding to a different antigen, were computationally redesigned to predict antibodies that could engage multiple antigens at once. The Rosetta multi-state design process predicted antibody sequences for the entire heavy chain variable region, including framework, CDR1, and CDR2 mutations. The predicted sequences matched the germline gene sequences to a remarkable degree, revealing by computational design the residues that are predicted to enable polyspecificity, i.e., binding of many unrelated antigens with a common sequence. The process thereby reverses antibody maturation in silico. In contrast, when designing antibodies to bind a single antigen, a sequence similar to that of the mature antibody sequence was returned, mimicking natural antibody maturation in silico. We demonstrated that the Rosetta computational design algorithm captures important aspects of antibody/antigen recognition. While the hypervariable region CDR3 often mediates much of the specificity of mature antibodies, we identified key positions in the VH gene encoding CDR1, CDR2, and the immunoglobulin framework that are critical contributors for polyspecificity in germline antibodies. Computational design of antibodies capable of binding multiple antigens may allow the rational design of antibodies that retain polyspecificity for diverse epitope binding. PMID:23637590

  2. HIV-1, HBV, HCV, HTLV, HPV-16/18, and Treponema pallidum Infections in a Sample of Brazilian Men Who Have Sex with Men

    PubMed Central

    Soares, Caroline C.; Georg, Ingebourg; Lampe, Elisabeth; Lewis, Lia; Morgado, Mariza G.; Nicol, Alcina F.; Pinho, Adriana A.; Salles, Regina C. S.; Teixeira, Sylvia L. M.; Vicente, Ana Carolina P.; Viscidi, Raphael P.; Gomes, Selma A.

    2014-01-01

    Background Men who have sex with men (MSM) are more vulnerable to blood-borne infections and/or sexually-transmitted infections (STI). This study was conducted to estimate the prevalences of mono and co-infections of HIV-1 and other blood-borne/STIs in a sample of MSM in Campinas, Brazil. Methods Responding Driven Sampling (RDS) was used for recruitment of MSM. Serum samples collected from 558 MSM were analyzed for the presence of serological markers for HIV-1, HBV, HCV, HTLV, HPV-16/18, and T. pallidum infections. Results The highest prevalences of infection in serum samples were found for HPV-16 and 18 (31.9% and 20.3%, respectively). Approximately 8% of the study population showed infection with HIV-1, and within that group, 27.5% had recently become infected with HIV-1. HBV infection and syphilis were detected in 11.4% and 10% of the study population, respectively, and the rates of HTLV and HCV infection were 1.5% and 1%, respectively. With the exception of HTLV, all other studied infections were usually found as co-infections rather then mono-infections. The rates of co-infection for HCV, HPV-18, and HIV-1 were the highest among the studied infections (100%, 83%, and 85%, respectively). Interestingly, HTLV infection was usually found as a mono-infection in the study group, whereas HCV was found only as a co-infection. Conclusions The present findings highlight the need to educate the MSM population concerning their risk for STIs infections and methods of prevention. Campaigns to encourage vaccination against HBV and HPV could decrease the rates of these infections in MSM. PMID:25083768

  3. Sandwich antibody arrays using recombinant antibody-binding protein L.

    PubMed

    Seo, Jin-soo; Poulter, C Dale

    2014-06-10

    Antibody arrays are a useful for detecting antigens and other antibodies. This technique typically requires a uniform and well-defined orientation of antibodies attached to a surface for optimal performance. A uniform orientation can be achieved by modification of antibodies to include a single site for attachment. Thus, uniformly oriented antibody arrays require a bioengineered modification for the antibodies directly immobilization on the solid surface. In this study, we describe a "sandwich-type" antibody array where unmodified antibodies are oriented through binding with regioselectively immobilized recombinant antibody-binding protein L. Recombinant proL-CVIA bearing C-terminal CVIA motif is post-translationally modified with an alkyne group by protein farnesyltransferase (PFTase) at the cysteine residue in the CVIA sequence to give proL-CVIApf, which is covalently attached to an azido-modified glass slide by a Huisgen [3 + 2] cycloaddition reaction. Slides bearing antibodies bound to slides coated with regioselectively immobilized proL-CVIApf gave stronger fluorescence outputs and those where the antibody-binding protein was immobilized in random orientations on an epoxy-modified slide. Properly selected capture and detection antibodies did not cross-react with immobilized proL-CVIApf in sandwich arrays, and the proL-CVIApf slides can be used for multiple cycles of detected over a period of several months. PMID:24841983

  4. Reshaping Antibody Diversity

    PubMed Central

    Wang, Feng; Ekiert, Damian C.; Ahmad, Insha; Yu, Wenli; Zhang, Yong; Bazirgan, Omar; Torkamani, Ali; Raudsepp, Terje; Mwangi, Waithaka; Criscitiello, Michael F.; Wilson, Ian A.; Schultz, Peter G.; Smider, Vaughn V.

    2014-01-01

    Summary Unlike humans or mice, some species have limited genome encoded combinatorial diversity potential, yet mount a robust antibody response. Cows are unusual in having exceptionally long CDR H3 loops and few V-regions, but the mechanism for creating diversity is not understood. Deep sequencing revealed that ultralong CDR H3s contain a remarkable complexity of cysteines, suggesting that disulfide-bonded mini-domains may arise during repertoire development. Indeed, crystal structures of two cow antibodies reveal that these CDR H3s form a very unusual architecture composed of a ?-strand “stalk” that supports a structurally diverse, disulfide-bonded, “knob” domain. Sequence analysis suggests that diversity arises from somatic hypermutation of an ultralong DH with a severe codon bias towards mutation to cysteine. These unusual antibodies can be elicited to recognize defined antigens through the knob domain. Thus, the bovine immune system produces an antibody repertoire composed of CDR H3s of unprecedented length that fold into a diversity of mini-domains generated through combinations of somatically generated disulfides. PMID:23746848

  5. Natural antibodies to glycans.

    PubMed

    Bovin, N V

    2013-07-01

    A wide variety of so-called natural antibodies (nAbs), i.e. immunoglobulins generated by B-1 cells, are directed to glycans. nAbs to glycans can be divided in three groups: 1) conservative nAbs, i.e. practically the same in all healthy donors with respect to their epitope specificity and level in blood; 2) allo-antibodies to blood group antigens; 3) plastic antibodies related to the first or the second group but discussed separately because their level changes considerably during diseases and some temporary conditions, in particular inflammation and pregnancy. Antibodies from the third group proved to be prospective markers of a number of diseases, whereas their unusual level (below or above the norm) is not necessarily the consequence of disease/state. Modern microarrays allowed the determination of the human repertoire, which proved to be unexpectedly broad. It was observed that the content of some nAbs reaches about 0.1% of total immunoglobulins. Immunoglobulins of M class dominate for most nAbs, constituting up to 80-90%. Their affinity (to a monovalent glycan, in KD terms) were found to be within the range 10(-4)-10(-6) M. Antibodies to Gal?1-3GlcNAc (Le(C)), 4-HSO3Gal?1-4GalNAc (4'-O-SuLN), Fuc?1-3GlcNAc, Fuc?1-4GlcNAc, GalNAc?1-3Gal (Adi), Gal?1-4Gal?1-4Glc (P(k)), Gal?1-4Gal?1-4GlcNAc (P1), GlcNAc?-terminated glycans, and hyaluronic acid should be noted among the nAbs revealed and studied during the last decade. At the same time, a kind of "taboo" is observed for a number of glycans: antibodies to Le(X) and Le(Y), and almost all gangliosides have not been observed in healthy persons. Many of the revealed nAbs were directed to constrained inner (core) part of glycan, directly adjoined to lipid of cell membrane or protein. The biological function of these nAbs remains unclear; for anti-core antibodies, a role of surveillance on appearance of aberrant, especially cancer, antigens is supposed. The first data related to oncodiagnostics based on quantitation of anti-glycan nAbs are reported. PMID:24010841

  6. Antibody mimetics: promising complementary agents to animal-sourced antibodies.

    PubMed

    Baloch, Abdul Rasheed; Baloch, Abdul Wahid; Sutton, Brian J; Zhang, Xiaoying

    2014-09-29

    Abstract Despite their wide use as therapeutic, diagnostic and detection agents, the limitations of polyclonal and monoclonal antibodies have inspired scientists to design the next generation biomedical agents, so-called antibody mimetics that offer many advantages over conventional antibodies. Antibody mimetics can be constructed by protein-directed evolution or fusion of complementarity-determining regions through intervening framework regions. Substantial progress in exploiting human, butterfly (Pieris brassicae) and bacterial systems to design and select mimetics using display technologies has been made in the past 10 years, and one of these mimetics [Kalbitor® (Dyax)] has made its way to market. Many challenges lie ahead to develop mimetics for various biomedical applications, especially those for which conventional antibodies are ineffective, and this review describes the current characteristics, construction and applications of antibody mimetics compared to animal-sourced antibodies. The possible limitations of mimetics and future perspectives are also discussed. PMID:25264572

  7. Targeting antibodies to the cytoplasm

    PubMed Central

    Marschall, Andrea L J; Frenzel, André; Schirrmann, Thomas; Schüngel, Manuela

    2011-01-01

    A growing number of research consortia are now focused on generating antibodies and recombinant antibody fragments that target the human proteome. A particularly valuable application for these binding molecules would be their use inside a living cell, e.g., for imaging or functional intervention. Animal-derived antibodies must be brought into the cell through the membrane, whereas the availability of the antibody genes from phage display systems allows intracellular expression. Here, the various technologies to target intracellular proteins with antibodies are reviewed. PMID:21099369

  8. [Antibody therapy for Alzheimer's disease].

    PubMed

    Tabira, Takeshi; Matsumoto, Shin-Ei; Jin, Haifeng

    2011-11-01

    In order to avoid Abeta-induced autoimmune encephalitis, several monoclonal and polyclonal antibodies are in clinical trials. These are bapineuzumab, solanezumab, ponezumab, gantenerumab, BAN2401, gammaguard and octagam. Since each antibody has a different antigen epitope of Abeta, anti-amyloid activities are different. It is unknown which antibody is effective for Alzheimer disease, and we must wait for the result of clinical trials. Some patients who developed tissue amyloid plaque immuno-reactive (TAPIR) antibody showed slower decline after AN-1792 vaccination. We developed TAPIR-like monoclonal antibody, which was found to react with Abeta oligomers preferentially. PMID:22277519

  9. Therapeutic antibodies against cancer

    PubMed Central

    Adler, Mark J.; Dimitrov, Dimiter S.

    2012-01-01

    Antibody-based therapeutics against cancer are highly successful in clinic and currently enjoy unprecedented recognition of their potential; 13 monoclonal antibodies (mAbs) have been approved for clinical use in the European Union and in the United States (one, mylotarg, was withdrawn from market in 2010). Three of the mAbs (bevacizumab, rituximab, trastuzumab) are in the top six selling protein therapeutics with sales in 2010 of more than $5 bln each. Hundreds of mAbs including bispecific mAbs and multispecific fusion proteins, mAbs conjugated with small molecule drugs and mAbs with optimized pharmacokinetics are in clinical trials. However, challenges remain and it appears that deeper understanding of mechanisms is needed to overcome major problems including resistance to therapy, access to targets, complexity of biological systems and individual variations. PMID:22520975

  10. Antibody-mediated radiotherapy

    SciTech Connect

    Bloomer, W.D.; Lipsztein, R.; Dalton, J.F.

    1985-05-01

    Antibodies that react with antigens on the surface of tumor cells but not normal cells have great potential for cancer detection and therapy. If radiolabeled without loss of immunologic specificity, such antibodies may be able to deliver cytoxic amounts of radiation. Target- cell specificity and a high extraction coefficient are necessary with any radionuclide in order to minimize normal tissue irradiation. Tumor- cell-retention time and the rate of catabolized radionuclide will also influence ultimate applicability. Among the unanswered questions for choosing a radionuclide is the choice of particle emitter. Although classic beta emitters have been used in a number of clinical situations, they have not had a major impact on disease outcome except in diseases of the thyroid. Unfortunately, Auger emitters such as iodine 125 are cytotoxic only when localized within close proximity to the genome. On the other hand, alpha emitters such as astatine 211 eliminate the need for subcellular sequestration but not cell-specific localization. 34 references.

  11. Antibodies for Immunohistochemistry

    Microsoft Academic Search

    Igor B. Buchwalow; Werner Böcker

    \\u000a The first use of the term Antikörper (the German word for antibody) occurred in a text by Paul Ehrlich (Fig. 1.1) in the conclusion of his article “Experimental Studies on Immunity,” published\\u000a in October 1891. Paul Ehrlich was born in 1854 in Strehlen (the German Province of Silesia, now in Poland). As a schoolboy\\u000a and student of medicine he was

  12. Antineutrophil cytoplasmic antibodies.

    PubMed

    Bosch, Xavier; Guilabert, Antonio; Font, Josep

    2006-07-29

    Much like other autoantibodies (eg, anti-double stranded DNA in systemic lupus erythematosus or antiglomerular basement membrane antibodies in Goodpasture's syndrome), antineutrophil cytoplasmic antibodies (ANCA) have provided doctors with a useful serological test to assist in diagnosis of small-vessel vasculitides, including Wegener's granulomatosis, microscopic polyangiitis, Churg-Strauss syndrome, and their localised forms (eg, pauci-immune necrotising and crescentic glomerulonephritis). 85-95% of patients with Wegener's granulomatosis, microscopic polyangiitis, and pauci-immune necrotising and crescentic glomerulonephritis have serum ANCA. ANCA directed to either proteinase 3 or myeloperoxidase are clinically relevant, yet the relevance of other ANCA remains unknown. Besides their diagnostic potential, ANCA might be valuable in disease monitoring. In addition, data seem to confirm the long-disputed pathogenic role of these antibodies. Present treatments for ANCA-associated vasculitis are not free from side-effects and as many as 50% of patients relapse within 5 years. Accurate understanding of the key pathogenic points of ANCA-associated vasculitis can undoubtedly provide a more rational therapeutic approach. PMID:16876669

  13. Microbials for the production of monoclonal antibodies and antibody fragments

    PubMed Central

    Spadiut, Oliver; Capone, Simona; Krainer, Florian; Glieder, Anton; Herwig, Christoph

    2014-01-01

    Monoclonal antibodies (mAbs) and antibody fragments represent the most important biopharmaceutical products today. Because full length antibodies are glycosylated, mammalian cells, which allow human-like N-glycosylation, are currently used for their production. However, mammalian cells have several drawbacks when it comes to bioprocessing and scale-up, resulting in long processing times and elevated costs. By contrast, antibody fragments, that are not glycosylated but still exhibit antigen binding properties, can be produced in microbial organisms, which are easy to manipulate and cultivate. In this review, we summarize recent advances in the expression systems, strain engineering, and production processes for the three main microbials used in antibody and antibody fragment production, namely Saccharomyces cerevisiae, Pichia pastoris, and Escherichia coli. PMID:24183828

  14. Antibody Production by Single Cells

    Microsoft Academic Search

    G. J. V. Nossal; Joshua Lederberg

    1958-01-01

    FAGREUS1 and others2,3 have shown that certain tissues from pre-sensitized animals can form antibody in vitro. This communication describes a technique whereby antibody production by single cells isolated in microdroplets can be detected. The technique is based on specific immobilization of Salmonella serotypes by anti-flagellar antibody. It was observed that single cells from a rat, simultaneously stimulated with two antigens,

  15. Human antibodies from transgenic animals

    Microsoft Academic Search

    Nils Lonberg

    2005-01-01

    Laboratory mice provide a ready source of diverse, high-affinity and high-specificity monoclonal antibodies (mAbs). However, development of rodent antibodies as therapeutic agents has been impaired by the inherent immunogenicity of these molecules. One technology that has been explored to generate low immunogenicity mAbs for in vivo therapy involves the use of transgenic mice expressing repertoires of human antibody gene sequences.

  16. Function-first antibody discovery

    PubMed Central

    Frendéus, Björn

    2013-01-01

    Therapeutic antibodies may mediate antineoplastic effects by altering the biological functions of their target, by directly stimulating the demise of cancer cells or by activating antibody-dependent immune effector mechanisms. We have recently provided in vivo proof-of-concept for a “function-first” target and drug discovery platform in which antibodies against a multitude of tumor-associated antigens are screened for biological effects in a target-unbiased manner. PMID:24083074

  17. Recovery and Purification of Antibody

    Microsoft Academic Search

    XueJun Han; Arthur Hewig; Ganesh Vedantham

    \\u000a Monoclonal antibody drugs have become a large portion of protein therapeutics and many antibody molecules are being evaluated\\u000a at various stages of clinical trials in the biopharmaceutical industry. This review article summarizes the state of the art\\u000a antibody purification techniques. The main focus is chromatographic techniques that include protein A, ion exchange and HIC.\\u000a For each technique, the mechanisms of

  18. A Monoclonal Antibody to Saxitoxin

    Microsoft Academic Search

    Rüdiger Hack; Volker Renz; Erwin Märtlbauer; Gerhard Terplan

    1990-01-01

    After immunization of six BALB\\/c mice with saxitoxin (STX) coupled to keyhole limpet hemocyanin only one mouse developed serum antibodies specific to STX. By hybridizing the splenocytes of this mouse with X63?Ag8.653 myeloma cells one hybridoma line secreting antibodies to STX was produced. The antibody was designated 5C2 and proved to be IgM. In an indirect enzyme immunoassay using STX

  19. Anti-flavin antibodies.

    PubMed

    Barber, M J; Eichler, D C; Solomonson, L P; Ackrell, B A

    1987-02-15

    Antibodies were elicited to FAD by using the hapten N-6-(6-aminohexyl)-FAD conjugated to the immunogenic carrier protein bovine serum albumin. Cross-reactivity was determined by Ouchterlony double-diffusion analysis with N-6-(6-aminohexyl)-FAD coupled to rabbit serum albumin. Anti-FAD IgG was partially purified by (NH4)2SO4 precipitation followed by DEAE-cellulose/CM-cellulose and bovine serum albumin-agarose chromatography. The partially purified anti-FAD IgG fraction failed to inhibit the catalytic activities of the flavin-containing enzymes nitrate reductase, xanthine oxidase and succinate dehydrogenase, whereas enzyme activity could be inhibited by addition of antibodies elicited against the native proteins. However, the partially purified anti-FAD IgG fraction could be used as a highly sensitive and specific probe to detect proteins containing only covalently bound flavin, such as succinate dehydrogenase, p-cresol methylhydroxylase and monoamine oxidase, by immuno-blotting techniques. Detection limits were estimated to be of the order of femtomolar concentrations of FAD with increased sensitivity for the 8 alpha-N(3)-histidyl linkage compared with 8 alpha-O-tyrosyl substitution. PMID:3109386

  20. STUDIES ON ANTIBODY PRODUCTION

    PubMed Central

    Ambrose, Charles T.; Coons, Albert H.

    1963-01-01

    When lymph node fragments from previously immunized rabbits were stimulated in vitro to produce a secondary response, the continuous presence of 50 µg/ml (0.15 mM) of chloramphenicol in the medium during the entire incubation period of 15 to 21 days produced nearly complete suppression of the response. Concentrations as low as 5 µg/ml (0.015 mM) produced approximately 80 per cent suppression of the response. When 50 µg/ml of chloramphenicol was present during only the first 6 days of culture, the secondary response was reduced 90 per cent. When it was absent for the first 6 days but present for the next 9 to 15 days, the response was reduced only 40 per cent. Since over 95 per cent of the antibody of the secondary response in most experiments appeared in the medium after the 6th day, chloramphenicol apparently inhibits antibody production by interfering with some early phase of the response. It is suggested that this interference involves messenger RNA and that animal cells have appeared resistant to this drug only because their complement of messenger RNA present when the drug has been added is stable over the short periods during which protein synthesis has usually been studied. PMID:14012518

  1. Radiolabeled antibodies in gynecologic tumors

    SciTech Connect

    Hardy, J.G.; Perkins, A.C.; Symonds, E.M.; Wastie, M.L.; Pimm, M.V.

    1984-01-01

    A monoclonal antibody has been raised against an osteogenic sarcoma cell line and radiolabeled with iodine-131. The antibody was administered to 12 patients with suspected ovarian tumors, two with recurrent carcinoma of the cervix and one with carcinoma of the body of the uterus. Each patient received an intravenous dose of 70 MBq I-131-labeled antibody and was imaged either 24 or 48 hours later. Image enhancement was achieved by subtraction of background activity using Tc-99m-labeled red blood cells and pertechnetate. In eleven patients with ovarian malignancies antibody uptake was detected at the suspected tumor sites, and agreed with the operative findings in the eight patients who subsequently underwent surgery. The patient in whom the antibody failed to localize was found to have a benign lesion. Uptake of antibody was seen at the tumor sites in the patients with carcinoma of the cervix and body of the uterus. The localization of tumor sites using I-131-labeled antibodies is difficult due to background activity, particularly from radioiodine in the bladder. In only five cases could the abnormal antibody concentration be identified on the iodine images alone. This problem was overcome by the use of background subtraction techniques. Immunoscintigraphy is proving useful for the assessment of tumor recurrence and as an aid to radiotherapy treatment planning.

  2. Renaissance of cancer therapeutic antibodies

    Microsoft Academic Search

    Martin J. Glennie

    2003-01-01

    In the past five years therapeutic monoclonal antibodies have established themselves as perhaps the most important and rapidly expanding class of therapeutic drugs. More than 25% of pharmacological agents that are currently under development are based on antibodies and the total income generated from them in 2002 exceeded $3 billion, and is predicted to rise to $10–20 billion by 2010.

  3. Endogenous Antibodies for Tumor Detection

    PubMed Central

    Rich, Barrie S.; Honeyman, Joshua N.; Darcy, David G.; Smith, Peter T.; Williams, Andrew R.; Lim, Irene Isabel P.; Johnson, Linda K.; Gönen, Mithat; Simon, Joel S.; LaQuaglia, Michael P.; Simon, Sanford M.

    2014-01-01

    The study of cancer immunology has provided diagnostic and therapeutic instruments through serum autoantibody biomarkers and exogenous monoclonal antibodies. While some endogenous antibodies are found within or surrounding transformed tissue, the extent to which this exists has not been entirely characterized. We find that in transgenic and xenograft mouse models of cancer, endogenous gamma immunoglobulin (IgG) is present at higher concentration in malignantly transformed organs compared to non-transformed organs in the same mouse or organs of cognate wild-type mice. The enrichment of endogenous antibodies within the malignant tissue provides a potential means of identifying and tracking malignant cells in vivo as they mutate and diversify. Exploiting these antibodies for diagnostic and therapeutic purposes is possible through the use of agents that bind endogenous antibodies. PMID:24875800

  4. RosettaAntibody: antibody variable region homology modeling server.

    PubMed

    Sircar, Aroop; Kim, Eric T; Gray, Jeffrey J

    2009-07-01

    The RosettaAntibody server (http://antibody.graylab.jhu.edu) predicts the structure of an antibody variable region given the amino-acid sequences of the respective light and heavy chains. In an initial stage, the server identifies and displays the most sequence homologous template structures for the light and heavy framework regions and each of the complementarity determining region (CDR) loops. Subsequently, the most homologous templates are assembled into a side-chain optimized crude model, and the server returns a picture and coordinate file. For users requesting a high-resolution model, the server executes the full RosettaAntibody protocol which additionally models the hyper-variable CDR H3 loop. The high-resolution protocol also relieves steric clashes by optimizing the CDR backbone torsion angles and by simultaneously perturbing the relative orientation of the light and heavy chains. RosettaAntibody generates 2000 independent structures, and the server returns pictures, coordinate files, and detailed scoring information for the 10 top-scoring models. The 10 models enable users to use rational judgment in choosing the best model or to use the set as an ensemble for further studies such as docking. The high-resolution models generated by RosettaAntibody have been used for the successful prediction of antibody-antigen complex structures. PMID:19458157

  5. Micromechanical antibody sensor

    DOEpatents

    Thundat, Thomas G. (Knoxville, TN); Jacobson, K. Bruce (Oak Ridge, TN); Doktycz, Mitchel J. (Knoxville, TN); Kennel, Stephen J. (Oak Ridge, TN); Warmack, Robert J. (Knoxville, TN)

    2001-01-01

    A sensor apparatus is provided using a microcantilevered spring element having a coating of a detector molecule such as an antibody or antigen. A sample containing a target molecule or substrate is provided to the coating. The spring element bends in response to the stress induced by the binding which occurs between the detector and target molecules. Deflections of the cantilever are detected by a variety of detection techniques. The microcantilever may be approximately 1 to 200 .mu.m long, approximately 1 to 50 .mu.m wide, and approximately 0.3 to 3.0 .mu.m thick. A sensitivity for detection of deflections is in the range of 0.01 nanometers.

  6. Encephalitis and GABAB receptor antibodies

    PubMed Central

    Höftberger, Romana; Titulaer, Maarten J.; Sabater, Lidia; Dome, Balazs; Rózsás, Anita; Hegedus, Balazs; Hoda, Mir Alireza; Laszlo, Viktoria; Ankersmit, Hendrik Jan; Harms, Lutz; Boyero, Sabas; de Felipe, Alicia; Saiz, Albert; Dalmau, Josep

    2013-01-01

    Objective: To report the clinical features of 20 newly diagnosed patients with GABAB receptor (GABABR) antibodies and determine the frequency of associated tumors and concurrent neuronal autoantibodies. Methods: Clinical data were retrospectively obtained and evaluated. Serum and CSF samples were examined for additional antibodies using methods previously reported. Results: Seventeen patients presented with seizures, memory loss, and confusion, compatible with limbic encephalitis (LE), one patient presented with ataxia, one patient presented with status epilepticus, and one patient presented with opsoclonus-myoclonus syndrome (OMS). Nineteen (95%) patients eventually developed LE during the course of the disease. Small-cell lung cancer (SCLC) was identified in 10 (50%) patients, all with LE. Treatment and outcome was available from 19 patients: 15 showed complete (n = 7) or partial (n = 8) neurologic improvement after steroids, IV immunoglobulins, or plasma exchange and oncologic treatment when indicated; 1 patient died of tumor progression shortly after the first cycle of immunotherapy, and 3 were not treated. Five patients with SCLC had additional onconeuronal antibodies (Ri, amphiphysin, or SOX1), and 2 without tumor had GAD65 and NMDAR antibodies, respectively. GABABR antibodies were not detected in serum of 116 patients with SCLC without neurologic symptoms. Conclusion: Our study confirms GABABR as an autoantigen of paraneoplastic and nonparaneoplastic LE and expands the phenotype of GABABR antibodies to ataxia, OMS, and status epilepticus. The long-term prognosis is dictated by the presence of a tumor. Recognition of syndromes associated with GABABR antibodies is important because they usually respond to treatment. PMID:24068784

  7. Modified antibody in fetal alloimmunization.

    PubMed

    Bussel, James B; McFarland, Janice G

    2013-07-18

    In this issue of Blood, Ghevaert et al propose to develop a therapeutic antibody for fetal and neonatal alloimmune thrombocytopenia (FNAIT) that would block the actual antibody in sensitized mothers from binding and therefore prevent, or at least ameliorate, fetal and neonatal thrombocytopenia in fetuses who would otherwise be affected.1 The goal of the group is to engineer an antibody reagent that would on the one hand not engage conventional activating Fc receptors and on the other hand interact normally with FcRn, allowing transplacental passage. PMID:23869072

  8. Reducing heterophilic antibody interference in immunoassays using single chain antibodies

    SciTech Connect

    Baird, Cheryl L.; Tan, Ruimin; Fischer, Christopher J.; Victry, Kristin D.; Zangar, Richard C.; Rodland, Karin D.

    2011-12-15

    Sandwich ELISA microarrays have the potential to simultaneously quantify the levels of multiple diagnostic targets in a biological sample. However, as seen with traditional ELISA diagnostics, heterophilic antibodies (HA) in patient sera have the potential to cause interference in these assays. We demonstrate here that reducing the diagnostic capture antibody to its minimal functional unit, the variable heavy and light domains artificially connected with a short polypeptide linker (scFv), is an effective strategy for reducing the HA assay interference.

  9. Structural Biology of Moonlighting --Lessons from Antibodies

    E-print Network

    Martin, Andrew C.R.

    Structural Biology of Moonlighting -- Lessons from Antibodies Andrew C.R. Martin Institute that they are both part of what an antibody needs to do. Nonetheless, antibodies provide interesting lessons fragment) forms the stem of the Y-shape and is responsible for the effector functions of the antibody

  10. Structural Biology of Moonlighting --Lessons from Antibodies

    E-print Network

    Martin, Andrew C.R.

    Structural Biology of Moonlighting -- Lessons from Antibodies Andrew C.R. Martin Institute that they are both part of what an antibody needs to do. Nonetheless, antibodies provide interesting lessons the stem of the Y-shape and is responsible for the effector functions of the antibody such as binding to F

  11. Surface chemistries for antibody microarrays

    SciTech Connect

    Seurynck-Servoss, Shannon L.; Baird, Cheryl L.; Rodland, Karin D.; Zangar, Richard C.

    2007-05-01

    Enzyme-linked immunosorbent assay (ELISA) microarrays promise to be a powerful tool for the detection of disease biomarkers. The original technology for printing ELISA microarray chips and capturing antibodies on slides was derived from the DNA microarray field. However, due to the need to maintain antibody structure and function when immobilized, surface chemistries used for DNA microarrays are not always appropriate for ELISA microarrays. In order to identify better surface chemistries for antibody capture, a number of commercial companies and academic research groups have developed new slide types that could improve antibody function in microarray applications. In this review we compare and contrast the commercially available slide chemistries, as well as highlight some promising recent advances in the field.

  12. Antiphospholipid-antibody-associated panniculitis.

    PubMed

    Hunt, Raegan D; Robinson, Maria; Patel, Rishi; Franks, Andrew G

    2012-12-01

    A 60-year-old man presented with intermittent, tender, erythematous nodules on the legs that were associated with mild arthralgias. He was otherwise asymptomatic but reported a history of lupus anticoagulant antibodies that were discovered incidentally on laboratory screening at the approximate time that his lesions first occurred. A biopsy specimen showed a septal and lobular panniculitis with neutrophils, histiocytes, numerous eosinophils, foci of fibrosis, and fat necrosis but no vascular pathology. An elevated activated partial thromboplastin time (PTT), appreciably elevated levels of anti-beta-2 glycoprotein I antibody (IgM and IgG), and moderately elevated levels of anticardiolipin antibody (IgM and IgG) were present. The onset and recurrence of his skin condition coincided with increased antiphospholipid antibody levels and treatment with 81 mg aspirin daily was associated with improvement. PMID:23286808

  13. Biophysical Signatures of Monoclonal Antibodies

    Microsoft Academic Search

    N. Harn; T. Spitznagel; M. Perkins; C. Allan; S. Shire; C. R. Middaugh

    \\u000a Monoclonal antibodies are the most common protein that is being developed by many companies as therapies against a wide range\\u000a of diseases (Andreakos et al. 2002; Campbell and Marcus 2003; Trikha et al. 2002; Untch et al. 2003). With increasing interest\\u000a in the use of monoclonal antibodies therapeutics, it is apparent that the combination of specificity and safety offered by

  14. Radioimmunoguided surgery using monoclonal antibody

    Microsoft Academic Search

    E. W. Jr. Martin; C. M. Mojzisik; G. H. Jr. Hinkle; J. Sampsel; M. A. Siddiqi; S. E. Tuttle; B. Sickle-Santanello; D. Colcher; M. O. Thurston; J. G. Bell

    1988-01-01

    The potential proficiency of radioimmunoguided surgery in the intraoperative detection of tumors was assessed using labeled monoclonal antibody B72.3 in 66 patients with tissue-proved tumor. Monoclonal antibody B72.3 was injected 5 to 42 days preoperatively, and the hand-held gamma-detecting probe was used intraoperatively to detect the presence of tumor. Intraoperative probe counts of less than 20 every 2 seconds, or

  15. Therapeutic Anti-VEGF Antibodies

    Microsoft Academic Search

    S. Lien; H. B. Lowman

    Vascular endothelial growth factor (VEGF-A) is a key cytokine in the development of normal blood vessels as well as the development\\u000a of vessels in tumors and other tissues undergoing abnormal angiogenesis. Here, we review the molecular engineering of two\\u000a humanized antibodies derived from a common mouse anti-VEGF antibody — bevacizumab, a full-length IgG1 approved for the treatment\\u000a of specified cancer

  16. Neutralising Antibodies against Ricin Toxin

    PubMed Central

    Prigent, Julie; Panigai, Laetitia; Lamourette, Patricia; Sauvaire, Didier; Devilliers, Karine; Plaisance, Marc; Volland, Hervé; Créminon, Christophe; Simon, Stéphanie

    2011-01-01

    The Centers for Disease Control and Prevention have listed the potential bioweapon ricin as a Category B Agent. Ricin is a so-called A/B toxin produced by plants and is one of the deadliest molecules known. It is easy to prepare and no curative treatment is available. An immunotherapeutic approach could be of interest to attenuate or neutralise the effects of the toxin. We sought to characterise neutralising monoclonal antibodies against ricin and to develop an effective therapy. For this purpose, mouse monoclonal antibodies (mAbs) were produced against the two chains of ricin toxin (RTA and RTB). Seven mAbs were selected for their capacity to neutralise the cytotoxic effects of ricin in vitro. Three of these, two anti-RTB (RB34 and RB37) and one anti-RTA (RA36), when used in combination improved neutralising capacity in vitro with an IC50 of 31 ng/ml. Passive administration of association of these three mixed mAbs (4.7 µg) protected mice from intranasal challenges with ricin (5 LD50). Among those three antibodies, anti-RTB antibodies protected mice more efficiently than the anti-RTA antibody. The combination of the three antibodies protected mice up to 7.5 hours after ricin challenge. The strong in vivo neutralising capacity of this three mAbs combination makes it potentially useful for immunotherapeutic purposes in the case of ricin poisoning or possibly for prevention. PMID:21633505

  17. Antibodies to watch in 2015.

    PubMed

    Reichert, Janice M

    2015-01-01

    The commercial pipeline of recombinant antibody therapeutics is robust and dynamic. As of early December 2014, a total of 6 such products (vedolizumab, siltuximab, ramucirumab, pembrolizumab, nivolumab, blinatumomab) were granted first marketing approvals in 2014. As discussed in this perspective on antibodies in late-stage development, the outlook for additional approvals, potentially still in 2014 and certainly in 2015, is excellent as marketing applications for 7 antibody therapeutics (secukinumab, evolocumab, mepolizumab, dinutuximab, nivolumab, blinatumomab, necitumumab) are undergoing a first regulatory review in the EU or US. Of the 39 novel mAbs currently in Phase 3 studies, a marketing application for one (alirocumab) may be submitted in late 2014, and marketing application submissions for at least 4 (reslizumab, ixekizumab, ocrelizumab, obiltoxaximab) are expected in 2015. Other 'antibodies to watch' are those in Phase 3 studies with estimated primary completion dates in late 2014 or 2015, which includes 13 for non-cancer indications (brodalumab, bimagrumab, bococizumab, MABp1, gevokizumab, dupilumab, sirukumab, sarilumab, tildrakizumab, guselkumab, epratuzumab, combination of actoxumab + bezlotoxumab, romosozumab) and 2 (racotumomab and clivatuzumab tetraxetan) undergoing evaluation as treatments for cancer. In addition to the novel antibody therapeutics mentioned, biosimilar infliximab and biosimilar trastuzumab are 'antibodies to watch' in 2015 because of their potential for entry into the US market and regulatory review, respectively. PMID:25484055

  18. Antibodies to watch in 2014.

    PubMed

    Reichert, Janice M

    2014-01-01

    Since 2010, mAbs has documented the biopharmaceutical industry's progress in transitioning antibody therapeutics to first Phase 3 clinical studies and regulatory review, and its success at gaining first marketing approvals for antibody-based products. This installment of the "Antibodies to watch" series outlines events anticipated to occur between December 2013 and the end of 2014, including first regulatory actions on marketing applications for vedolizumab, siltuximab, and ramucirumab, as well as the Fc fusion proteins Factor IX-Fc and Factor VIII-Fc; and the submission of first marketing applications for up to five therapeutics (secukinumab, ch14.18, onartuzumab, necitumumab, gevokizumab). Antibody therapeutics in Phase 3 studies are described, with an emphasis on those with study completion dates in 2014, including antibodies targeting interleukin-17a or the interleukin-17a receptor (secukinumab, ixekizumab, brodalumab), proprotein convertase subtilisin/kexin type 9 (alirocumab, evolocumab, bococizumab), and programmed death 1 receptor (lambrolizumab, nivolumab). Five antibodies with US Food and Drug Administration's Breakthrough Therapy designation (obinutuzumab, ofatumumab, lambrolizumab, bimagrumab, daratumumab) are also discussed. PMID:24284914

  19. Antibodies to watch in 2014

    PubMed Central

    Reichert, Janice M

    2014-01-01

    Since 2010, mAbs has documented the biopharmaceutical industry’s progress in transitioning antibody therapeutics to first Phase 3 clinical studies and regulatory review, and its success at gaining first marketing approvals for antibody-based products. This installment of the “Antibodies to watch” series outlines events anticipated to occur between December 2013 and the end of 2014, including first regulatory actions on marketing applications for vedolizumab, siltuximab, and ramucirumab, as well as the Fc fusion proteins Factor IX-Fc and Factor VIII-Fc; and the submission of first marketing applications for up to five therapeutics (secukinumab, ch14.18, onartuzumab, necitumumab, gevokizumab). Antibody therapeutics in Phase 3 studies are described, with an emphasis on those with study completion dates in 2014, including antibodies targeting interleukin-17a or the interleukin-17a receptor (secukinumab, ixekizumab, brodalumab), proprotein convertase subtilisin/kexin type 9 (alirocumab, evolocumab, bococizumab), and programmed death 1 receptor (lambrolizumab, nivolumab). Five antibodies with US Food and Drug Administration’s Breakthrough Therapy designation (obinutuzumab, ofatumumab, lambrolizumab, bimagrumab, daratumumab) are also discussed. PMID:24284914

  20. Elimination mechanisms of therapeutic monoclonal antibodies.

    PubMed

    Tabrizi, Mohammad A; Tseng, Chih-Ming L; Roskos, Lorin K

    2006-01-01

    Targeted therapies using monoclonal antibodies have achieved important therapeutic applications in the treatment of various human diseases. Understanding the factors that impact the pharmacokinetics of monoclonal antibodies is of high importance for effective therapy. Many factors related to the target antigen, antibody and patients can affect antibody elimination. Evaluation of these factors will facilitate the understanding of the processes involved in antibody elimination. PMID:16478695

  1. Pharmacokinetic characteristics of therapeutic antibodies.

    PubMed

    Wohlrab, Johannes

    2015-06-01

    Because of their high molecular weight and their highly hydrophilic character, therapeutic antibodies behave differently in terms of absorption, distribution and elimination compared to conventional drugs. Also, their pharmacokinetic profile varies significantly among individuals. After subcutaneous administration, antibodies are absorbed via the lymphatic system and become systemically bioavailable with some delay. The physicochemical properties of the molecules hinder their distribution from the bloodstream into the tissue. Elimination occurs by proteolysis in various organs (skin, muscle, liver), but mainly within the reticuloendothelial system. Also relevant is the elimination through target antigens (especially in the case of cell-bound target antigens) as well as a recycling process through binding to the neonatal Fc receptor that provides protection from lysosomal degradation. Depending on the immunogenicity of the therapeutic antibody and the individual immune response, neutralizing antibodies can develop. Pharmacokinetic conditions can be optimized by coadministration of, for example, methotrexate. Moreover, risk factors for the loss of immunological tolerance, such as on-demand therapy or elective switching of therapeutic antibodies, should be avoided. PMID:26018364

  2. Monoclonal Antibodies for the Treatment of Cancer

    PubMed Central

    Shuptrine, Casey; Surana, Rishi; Weiner, Louis M.

    2012-01-01

    Over the past decade, the clinical utility of monoclonal antibodies has been realized and antibodies are now a mainstay for the treatment of cancer. Antibodies have the unique capacity to target and kill tumor cells while simultaneously activating immune effectors to kill tumor cells through the complement cascade or antibody-dependent cellular cytotoxicity (ADCC). This multifaceted mechanism of action combined with target specificity underlies the capacity of antibodies to elicit anti-tumor responses while minimizing the frequency and magnitude of adverse events. This review will focus on mechanisms of action, clinical applications and putative mechanisms of resistance to monoclonal antibody therapy in the context of cancer. PMID:22245472

  3. Monoclonal Antibodies in Diagnosis and Therapy

    NASA Astrophysics Data System (ADS)

    Waldmann, Thomas A.

    1991-06-01

    Monoclonal antibodies have been applied clinically to the diagnosis and therapy of an array of human disorders, including cancer and infectious diseases, and have been used for the modulation of immune responses. Effective therapy using unmodified monoclonal antibodies has, however, been elusive. Recently, monoclonal antibody-mediated therapy has been revolutionized by advances such as the definition of cell-surface structures on abnormal cells as targets for effective monoclonal antibody action, genetic engineering to create less immunogenic and more effective monoclonal antibodies, and the arming of such antibodies with toxins or radionuclides to enhance their effector function.

  4. Controlling chemical reactivity with antibodies

    SciTech Connect

    Hsieh, L.C.; Schultz, P.G. (Univ. of California, Berkeley (United States)); Yonkovich, S.; Kochersperger, L. (Affymax Research Inst., Palo Alto, CA (United States))

    1993-04-16

    The remarkable specificity of an antibody molecule has been used to accomplish highly selective functional group transformations not attainable by current chemical methods. An antibody raised against an amine-oxide hapten catalyzes the reduction of a diketone to a hydroxyketone with greater than 75:1 regioselectivity for one of two nearly equivalent ketone moieties. The antibody-catalyzed reaction is highly stereoselective, affording the hydroxyketone in high enantiomeric excess. Similarly, the reduction of ketones containing branched and aryl substituents, including the highly symmetrical 1-nitrophenyl-3-phenyl-2-propanone, was enantioselective. The simple strategy presented herein may find general applicability to the regio- and stereoselective reduction of a broad range of compounds. 18 refs., 2 figs., 1 tab.

  5. Molecular-specific urokinase antibodies

    NASA Technical Reports Server (NTRS)

    Atassi, M. Zouhair (Inventor); Morrison, Dennis R. (Inventor)

    2009-01-01

    Antibodies have been developed against the different molecular forms of urokinase using synthetic peptides as immunogens. The peptides were synthesized specifically to represent those regions of the urokinase molecules which are exposed in the three-dimensional configuration of the molecule and are uniquely homologous to urokinase. Antibodies are directed against the lysine 158-isoleucine 159 peptide bond which is cleaved during activation from the single-chain (ScuPA) form to the bioactive double chain (54 KDa and 33 KDa) forms of urokinase and against the lysine 135 lysine 136 bond that is cleaved in the process of removing the alpha-chain from the 54 KDa form to produce the 33 KDa form of urokinase. These antibodies enable the direct measurement of the different molecular forms of urokinase from small samples of conditioned medium harvested from cell cultures.

  6. Antibodies to watch in 2013

    PubMed Central

    Reichert, Janice M

    2013-01-01

    The transitions of antibody therapeutics to late-stage clinical development, regulatory review and the market are proceeding at a rapid pace in 2013. Since late 2012, two monoclonal antibody (mAb) therapeutics (itolizumab, trastuzumab emtansine) received their first approvals, first marketing applications for three mAbs (vedolizumab, ramucirumab, obinutuzumab) were submitted to regulatory agencies, and five mAbs (brodalumab, MABp1, moxetumomab pasudotox, tildrakizumab, rilotumumab) entered their first Phase 3 studies. The current total of commercially-sponsored antibody therapeutics undergoing evaluation in late-stage studies is 30. Recently announced study results for farletuzumab, naptumomab estafenatox, and tabalumab indicate that clinical endpoints were not met in some Phase 3 studies of these product candidates. PMID:23727858

  7. Entanglement model of antibody viscosity.

    PubMed

    Schmit, Jeremy D; He, Feng; Mishra, Shradha; Ketchem, Randal R; Woods, Christopher E; Kerwin, Bruce A

    2014-05-15

    Antibody solutions are typically much more viscous than solutions of globular proteins at equivalent volume fraction. Here we propose that this is due to molecular entanglements that are caused by the elongated shape and intrinsic flexibility of antibody molecules. We present a simple theory in which the antibodies are modeled as linear polymers that can grow via reversible bonds between the antigen binding domains. This mechanism explains the observation that relatively subtle changes to the interparticle interaction can lead to large changes in the viscosity. The theory explains the presence of distinct power law regimes in the concentration dependence of the viscosity as well as the correlation between the viscosity and the charge on the variable domain in our antistreptavidin IgG1 model system. PMID:24758234

  8. Antibody microarrays utilizing site-specific antibody-oligonucleotide conjugates.

    PubMed

    Wold, Erik D; McBride, Ryan; Axup, Jun Y; Kazane, Stephanie A; Smider, Vaughn V

    2015-05-20

    Protein arrays are typically made by random absorption of proteins to the array surface, potentially limiting the amount of properly oriented and functional molecules. We report the development of a DNA encoded antibody microarray utilizing site-specific antibody-oligonucleotide conjugates that can be used for cell immobilization as well as the detection of genes and proteins. This technology allows for the facile generation of antibody microarrays while circumventing many of the drawbacks of conventionally produced antibody arrays. We demonstrate that this method can be used to capture and detect SK-BR-3 cells (Her2+ breast cancer cells) at concentrations as low as 10(2) cells/mL (which is equivalent to 10 cells per 100 ?L array) without the use of microfluidics, which is 100- to 10(5)-fold more sensitive than comparable techniques. Additionally, the method was shown to be able to detect cells in a complex mixture, effectively immobilizing and specifically detecting Her2+ cells at a concentration of 10(2) SK-BR-3 cells/mL in 4 × 10(6) white blood cells/mL. Patients with a variety of cancers can have circulating tumor cell counts of between 1 and 10(3) cells/mL in whole blood, well within the range of this technology. PMID:25884500

  9. Antibody profiling sensitivity through increased reporter antibody layering

    DOEpatents

    Apel, William A.; Thompson, Vicki S.

    2013-02-26

    A method for analyzing a biological sample by antibody profiling for identifying forensic samples or for detecting the presence of an analyte. In an embodiment of the invention, the analyte is a drug, such as marijuana, Cocaine (crystalline tropane alkaloid), methamphetamine, methyltestosterone, or mesterolone. The method comprises attaching antigens to a surface of a solid support in a preselected pattern to form an array wherein locations of the antigens are known; contacting the array with the biological sample such that a portion of antibodies in the sample reacts with and binds to the antigens in the array to form immune complexes; washing away antibodies that do form immune complexes; and detecting the immune complexes, to form an antibody profile. Forensic samples are identified by comparing a sample from an unknown source with a sample from a known source. Further, an assay, such as a test for illegal drug use, can be coupled to a test for identity such that the results of the assay can be positively correlated to the subject's identity.

  10. Antibody profiling sensitivity through increased reporter antibody layering

    DOEpatents

    Apel, William A.; Thompson, Vicki S

    2010-04-13

    A method for analyzing a biological sample by antibody profiling for identifying forensic samples or for detecting the presence of an analyte. In an embodiment of the invention, the analyte is a drug, such as marijuana, Cocaine (crystalline tropane alkaloid), methamphetamine, methyltestosterone, or mesterolone. The method comprises attaching antigens to a surface of a solid support in a preselected pattern to form an array wherein locations of the antigens are known; contacting the array with the biological sample such that a portion of antibodies in the sample reacts with and binds to the antigens in the array to form immune complexes; washing away antibodies that do form immune complexes; and detecting the immune complexes, to form an antibody profile. Forensic samples are identified by comparing a sample from an unknown source with a sample from a known source. Further, an assay, such as a test for illegal drug use, can be coupled to a test for identity such that the results of the assay can be positively correlated to the subject's identity.

  11. [Antineutrophil cytoplasmic antibodies and associated diseases].

    PubMed

    Sghiri, R; Meddeb, H; Bouguila, J; Jarray, M; Bahri, F; Nouira, R; Zellama, D; Achour, A; Essoussi, A S; Harbi, A; Ghedira, I

    2009-07-01

    Antineutrophil cytoplasmic antibodies are classical serological markers of small-vessels vasculitis. However, they have been described in many other pathological situations. The aim of this study was to determine through our experience, the main antineutrophil cytoplasmic antibodies-associated diseases and to investigate antigen targets of these antibodies. Forty complete observations of antineutrophil cytoplasmic antibodies (ANCA) positive patients either by indirect immunofluorescence or by enzyme immunoassay were analysed. Only five (12.5%) patients have small-vessels vasculitis. Among these, antineutrophil cytoplasmic antibodies were detected only by Elisa in one patient and they were exclusively directed against bactericidal permeability increasing protein in another one. Our study confirms the presence of antineutrophil cytoplasmic antibodies in different diseases. It demonstrates that antineutrophil cytoplasmic antibodies should be investigated by Elisa when indirect immunofluorescence is negative. In small-vessels vasculitis, Proteinase 3 and myeloperoxidase are mainly but not exclusively the antigenic targets of antineutrophil cytoplasmic antibodies. PMID:18834675

  12. Detection of Campylobacter species using monoclonal antibodies

    NASA Astrophysics Data System (ADS)

    Young, Colin R.; Lee, Alice; Stanker, Larry H.

    1999-01-01

    A panel of species specific monoclonal antibodies were raised to Campylobacter coli, Campylobacter jejuni and Campylobacter lari. The isotypes, and cross-reactivity profiles of each monoclonal antibody against an extensive panel of micro- organisms, were determined.

  13. Anti-insulin antibody test

    MedlinePLUS

    Insulin antibodies - serum; Insulin Ab test ... This test may be performed if you have or are at risk for type 1 diabetes . It also may ... different laboratories. Some labs use different measurements or test different samples. Talk to your doctor about the ...

  14. Anti-acetylcholine receptor antibodies.

    PubMed Central

    Vincent, A; Newsom Davis, J

    1980-01-01

    Early suggestions that a humoral factor might be implicated in the disorder of neuromuscular transmission in myasthenia gravis have been confirmed by the detection of anti-AChR antibody in 85-90% of the patients with generalised disease and in 75% of cases with restricted ocular myasthenia. Plasma exchange reveals that serum anti-AChR usually has an inverse relationship to muscle strength and present evidence indicates that patients responding to thymectomy and immunosuppressive durg treatment usually show a consistent decline in serum anti-AChR titres. The antibody is heterogeneous and can lead to a loss of muscle AChR by several mechanisms. Anti-AChR is produced in the thymus in relatively small amounts. Anti-AChR antibody synthesis by thymic lymphocytes and pokeweed stimulated peripheral lymphocytes in culture provides a means of studying the effect of different lymphocyte populations in vitro. Analysis of clinical, immunological and HLA antigen characteristics in MG suggest that more than one mechanism may underlie the breakdown in tolerance to AChR, leading to the production of anti-AChR antibodies. PMID:7400823

  15. Therapeutic antibodies for autoimmunity and inflammation

    Microsoft Academic Search

    Andrew C. Chan; Paul J. Carter

    2010-01-01

    The development of therapeutic antibodies has evolved over the past decade into a mainstay of therapeutic options for patients with autoimmune and inflammatory diseases. Substantial advances in understanding the biology of human diseases have been made and tremendous benefit to patients has been gained with the first generation of therapeutic antibodies. The lessons learnt from these antibodies have provided the

  16. Monoclonal antibodies as neural cell surface markers

    Microsoft Academic Search

    O. K. Langley; M. S. Ghandour; G. Gombos; M. Hirn; C. Goridis

    1982-01-01

    A comparison is made of the immunohistochemistry at the ultrastructural level of three monoclonal antibodies directed against surface components of CNS cells. Hybridomas secreting these antibodies were obtained from two cell fusions of a rat myeloma cell line and immune splenocytes derived from rats immunized either with primary mouse brain cultured cells or membrane components. In cultures one antibody, anti-BSP-2

  17. Anti-DNA antibodies in SLE

    SciTech Connect

    Voss, E.W.

    1988-01-01

    This book contains 8 chapters. Some of the titles are: Anti-DNA Antibodies in SLE: Historical Perspective; Specificity of Anti-DNA Antibodies in Systemic Lupus Erythematosus; Monoclonial Autoimmune Anti-DNA Antibodies; and Structure--Function Analyses of Anti-DNA Autoantibodies.

  18. Review on modeling anti-antibody responses to monoclonal antibodies.

    PubMed

    Gómez-Mantilla, José David; Trocóniz, Ińaki F; Parra-Guillén, Zinnia; Garrido, María J

    2014-10-01

    Monoclonal antibodies (mAbs) represent a therapeutic strategy that has been increasingly used in different diseases. mAbs are highly specific for their targets leading to induce specific effector functions. Despite their therapeutic benefits, the presence of immunogenic reactions is of growing concern. The immunogenicity identified as anti-drug antibodies (ADA) production due to the continuous administration of mAbs may affect the pharmacokinetics (PK) and/or the pharmacodynamics (PD) of mAbs administered to patients. Therefore, the immunogenicity and its clinical impact have been studied by several authors using PK modeling approaches. In this review, the authors try to present all those models under a unique theoretical mechanism-based framework incorporating the main considerations related to ADA formation, and how ADA may affect the efficacy or toxicity profile of some therapeutic biomolecules. PMID:25027160

  19. Development of Broadly Neutralizing Antibodies from Autologous Neutralizing Antibody Responses

    PubMed Central

    Derdeyn, Cynthia A.; Moore, Penny L.; Morris, Lynn

    2014-01-01

    Purpose of Review Detailed genetic and structural characterization has revealed that broadly neutralizing antibodies (bnAbs) against HIV-1 have unusually high levels of somatic hypermutation, long CDRH3 domains, and the ability to target one of four sites of vulnerability on the HIV-1 envelope (Env) glycoproteins. A current priority is to understand how bnAbs are generated during natural infection, and translate this information into immunogens that can elicit bnAb following vaccination. Recent Findings Strain-specific neutralizing antibodies (nAb) can acquire broad neutralizing capacity when the transmitted/founder Env or a specific Env variant is recognized by an unmutated rearranged germline that has the capacity to develop bnAb like features. This could be a relatively infrequent event, as only certain germlines appear to possess inherent features needed for bnAb activity. Furthermore, the glycosylation pattern and diversity of circulating HIV-1 Envs, as well as the state of the B cell compartment, may influence the activation and maturation of certain antibody lineages. Summary Collectively, studies over the last year suggest that the development of HIV-1 Env immunogens that bind and activate bnAb-like germlines is feasible. However, more information about the features of Env variants and the host factors that lead to breadth during natural infection is needed to elicit bnAbs through immunization. PMID:24662931

  20. Engineered antibodies accelerating drug discovery and development.

    PubMed

    Zhu, Z

    2001-12-01

    This meeting, organized by the Strategic Research Institute, reviewed advances in both engineering technologies and clinical development of antibody products. A panel of speakers, mainly from the biotechnology and pharmaceutical industries, covered several important issues in the development of antibody therapeutics, including new display technologies, human antibody generation, high-level expression of antibodies in both mammalian cells and in transgenic plants, and the latest data from ongoing clinical trials of antibody products. In the case of human antibody generation, Abgenix Inc's XenoMax technology combines the powers of the transgenic XenoMouse and in vitro B cell culture and selection to enable the robust identification of antibodies with rare properties and very high affinity. MAbstract from Crucell NV provides a unique approach to the identification of human antibodies directed against tumor-specific glycosylation variants and activation epitopes of otherwise normal cellular molecules. Regarding antibody production, the use of transgenic plants is gaining more interest among biopharmaceutical industrials. Promising results were also reported on several antibody-based therapeutic, including antibody-drug conjugates and prodrugs. PMID:15931562

  1. Expression studies of catalytic antibodies.

    PubMed Central

    Ulrich, H D; Patten, P A; Yang, P L; Romesberg, F E; Schultz, P G

    1995-01-01

    We have examined the positive influence of human constant regions on the folding and bacterial expression of active soluble mouse immunoglobulin variable domains derived from a number of catalytic antibodies. Expression yields of eight hybridoma- and myeloma-derived chimeric Fab fragments are compared in both shake flasks and high density fermentations. In addition the usefulness of this system for the generation of in vivo expression libraries is examined by constructing and expressing combinations of heavy and light chain variable regions that were not selected as a pair during an immune response. A mutagenesis study of one of the recombinant catalytic Fab fragments reveals that single amino acid substitutions can have dramatic effects on the expression yield. This system should be generally applicable to the production of Fab fragments of catalytic and other hybridoma-derived antibodies for crystallographic and structure-function studies. Images Fig. 3 Fig. 4 PMID:8524873

  2. Antinuclear antibodies in rosacea patients

    PubMed Central

    Salamon, Ma?gorzata; McCauliffe, Daniel; Sysa-J?drzejowska, Anna

    2013-01-01

    Introduction Rosacea is a common inflammatory disorder, characterized by a spectrum of facial manifestations. The clinical similarity to other dermatoses, like lupus erythematosus, might lead to misdiagnosis, particularly in patients with elevated antinuclear antibody titers. Aim To assess the frequency, titer and specificity of antinuclear antibodies in rosacea patients and correlate these findings with clinical features. Material and methods The study included 101 rosacea patients and 26 sex- and age-matched controls. Immunofluorescence antinuclear antibody testing was performed on HEp-2 substrates. Patients’ sera with ANA titers of 1 : 160 or higher were evaluated by Euroline analysis. Results Over a half (53.5%) of rosacea patients had an ANA titer greater than or equal to 1 : 160. Within this group 13.86% had a titer of 1 : 320, 8.91% had a titer of 1 : 640, and 6.93% had a titer of 1 : 1,280 or higher. The specificity of these antibodies could not be identified. Elevated ANA titers were present more often in women (55.8%) than in men (44.15%). Only two of 26 healthy volunteers had elevated ANA titers. One had a titer of 1 : 160 and the other of 1 : 320. During a two-year observation period, after the initial ANA testing, none of the patients with ANA titers above 1 : 640 developed an apparent autoimmune disorder. Conclusions Elevated ANA titers are commonly found in rosacea patients, what with simultaneously existing facial erythema and photosensitivity might lead to misdiagnosis of lupus erythematosus. Clinicians should beware of these findings to avoid misdiagnosing lupus erythematosus in rosacea patients with elevated ANA titers. PMID:24278039

  3. Antiphospholipid antibody effects on monocytes

    Microsoft Academic Search

    Alisa S. Wolberg

    2007-01-01

    Although the presence of autoantibodies is known to increase the risk of thrombosis in the antiphospholipid syndrome, the\\u000a mechanism by which these antibodies exert their effects is poorly understood. Several studies suggest that autoantibody-mediated\\u000a dysregulation of monocytes is one pathobiologic mechanism of this disease. Recent studies have focused on extra-and intracellular\\u000a interactions involved in monocyte activation and expression of procoagulant

  4. Cytokine-neutralizing therapeutic antibodies

    Microsoft Academic Search

    Amanda Suitters; Roly Foulkes

    \\u000a Monoclonal antibodies (Mabs) offer the potential as useful therapeutic agents because of their high affinity and selectivity\\u000a for the target antigen. The standard approach in making Mabs has been to immunise rodents, usually mice, with the desired\\u000a human protein and generate a mouse anti-human IgG through hybridoma technology. Indeed mouse Mabs are used clinically in acute\\u000a diseases, such as the

  5. Therapeutic monoclonal antibodies in ophthalmology

    Microsoft Academic Search

    Eduardo B. Rodrigues; Michel E. Farah; Maurício Maia; Fernando M. Penha; Caio Regatieri; Gustavo B. Melo; Marcelo M. Pinheiro; Carlos R. Zanetti

    2009-01-01

    Monoclonal antibodies (mAbs) can be used therapeutically by binding to molecular targets with high specificity. Therefore, they have excellent therapeutic applications in ophthalmology. This manuscript presents four aspects of the therapeutic use of mAbs in ophthalmology: the scientific rationale, the unique characteristics of selected mAbs, the current state-of-the-art application, and relevant therapeutic mAbs for future applications in ophthalmology. We identified

  6. Bispecific Antibodies for Diagnostic Applications

    Microsoft Academic Search

    Archana Parashar; Susmita Sarkar; Advaita Ganguly; Sai Kiran Sharma; Mavanur R. Suresh

    \\u000a Bispecific monoclonal antibodies (BsMAb) are unique engineered macromolecules that have two different pre-determined binding\\u000a specificities. Their ability to simultaneously bind to a specific antigen and a given detection moiety enables them to function\\u000a as excellent bifunctional immunoprobes in diagnostic assays. BsMAb are being exploited for the development of simple, rapid,\\u000a and highly sensitive immunoassays for diagnosis of bacterial and viral

  7. Antibodies to laminin in Chagas' disease

    PubMed Central

    1982-01-01

    We have found that sera from humans with Chagas' disease and Rhesus monkeys infected with Trypanosoma cruzi contain IgM and IgG antibodies, which react with structures in a variety of connective tissues. These antibodies react with laminin but not with various other purified connective tissue components like collagen types I, III, IV, and V, fibronectin, heparan sulfate (BM-1) proteoglycan, or chondronectin. The tissue-reacting antibodies were isolated by absorption to a laminin- Sepharose column. The bound fraction contained all the tissue-reacting antibodies. These antibodies strongly stained trypomastigotes and amastigotes, but weakly stained epimastigotes. These studies show that sera from T. cruzi-infected primates contain antilaminin antibodies, which may be produced by those host in response to a laminin-like molecule present in the parasite. PMID:6801186

  8. Antibody to dihydropyridine calcium entry blockers

    SciTech Connect

    Thayer, S.; Minaskanian, G.; Fairhurst, A.

    1986-03-05

    Antibodies that recognize dihydropyridine calcium entry blockers were elicited from rabbits. A sensitive and specific radioimmunoassay for dihydropyridines was developed and its specificity compared to the DHP binding site in skeletal muscle membranes. The antibody bound (/sup 3/H)nitrendipine with a higher affinity (K/sub D/ = 0.155 nM) than did the DHP receptor of skeletal muscle (K/sub D/ = 1-3 nM). However, in contrast to the DHP receptor, the antibody recognized only those DHP drugs with meta-nitrophenyl substituents at the 4-position on the DHP ring, and thus reflected the meta position of the nitro group on the DHP hapten used as an antigen. Both the antibody and the receptor exhibited stereospecificity, with each site recognizing the (+) isomer of nicardipine as the more potent. This antibody should prove useful in the studies of some potentially irreversible DHP molecules and for use in the production of anti-idotype antibodies.

  9. Gangliosides, Ab1 and Ab2 antibodies

    Microsoft Academic Search

    Alejandro López-Requena; Marco Bestagno; Cristina Mateo de Acosta; Michela Cesco-Gaspere; Ana María Vázquez; Rolando Pérez; Oscar R. Burrone

    2007-01-01

    P3 mAb is an IgM monoclonal antibody specific for N-glycolyl-containing gangliosides. The immunogenicity of the P3 idiotype has been previously described by immunizing syngeneic BALB\\/c mice with the purified murine IgM or the mouse-human chimeric IgG antibody. In the present work we study the antibody response against the idiotype of P3 mAb through immunization with DNA. We used small immune

  10. Reconciling the Structural Attributes of Avian Antibodies*

    PubMed Central

    Conroy, Paul J.; Law, Ruby H. P.; Gilgunn, Sarah; Hearty, Stephen; Caradoc-Davies, Tom T.; Lloyd, Gordon; O'Kennedy, Richard J.; Whisstock, James C.

    2014-01-01

    Antibodies are high value therapeutic, diagnostic, biotechnological, and research tools. Combinatorial approaches to antibody discovery have facilitated access to unique antibodies by surpassing the diversity limitations of the natural repertoire, exploitation of immune repertoires from multiple species, and tailoring selections to isolate antibodies with desirable biophysical attributes. The V-gene repertoire of the chicken does not utilize highly diverse sequence and structures, which is in stark contrast to the mechanism employed by humans, mice, and primates. Recent exploitation of the avian immune system has generated high quality, high affinity antibodies to a wide range of antigens for a number of therapeutic, diagnostic and biotechnological applications. Furthermore, extensive examination of the amino acid characteristics of the chicken repertoire has provided significant insight into mechanisms employed by the avian immune system. A paucity of avian antibody crystal structures has limited our understanding of the structural consequences of these uniquely chicken features. This paper presents the crystal structure of two chicken single chain fragment variable (scFv) antibodies generated from large libraries by phage display against important human antigen targets, which capture two unique CDRL1 canonical classes in the presence and absence of a non-canonical disulfide constrained CDRH3. These structures cast light on the unique structural features of chicken antibodies and contribute further to our collective understanding of the unique mechanisms of diversity and biochemical attributes that render the chicken repertoire of particular value for antibody generation. PMID:24737329

  11. Reconciling the structural attributes of avian antibodies.

    PubMed

    Conroy, Paul J; Law, Ruby H P; Gilgunn, Sarah; Hearty, Stephen; Caradoc-Davies, Tom T; Lloyd, Gordon; O'Kennedy, Richard J; Whisstock, James C

    2014-05-30

    Antibodies are high value therapeutic, diagnostic, biotechnological, and research tools. Combinatorial approaches to antibody discovery have facilitated access to unique antibodies by surpassing the diversity limitations of the natural repertoire, exploitation of immune repertoires from multiple species, and tailoring selections to isolate antibodies with desirable biophysical attributes. The V-gene repertoire of the chicken does not utilize highly diverse sequence and structures, which is in stark contrast to the mechanism employed by humans, mice, and primates. Recent exploitation of the avian immune system has generated high quality, high affinity antibodies to a wide range of antigens for a number of therapeutic, diagnostic and biotechnological applications. Furthermore, extensive examination of the amino acid characteristics of the chicken repertoire has provided significant insight into mechanisms employed by the avian immune system. A paucity of avian antibody crystal structures has limited our understanding of the structural consequences of these uniquely chicken features. This paper presents the crystal structure of two chicken single chain fragment variable (scFv) antibodies generated from large libraries by phage display against important human antigen targets, which capture two unique CDRL1 canonical classes in the presence and absence of a non-canonical disulfide constrained CDRH3. These structures cast light on the unique structural features of chicken antibodies and contribute further to our collective understanding of the unique mechanisms of diversity and biochemical attributes that render the chicken repertoire of particular value for antibody generation. PMID:24737329

  12. Specificity of secretory antibodies to bacterial immunogens.

    PubMed

    Ebersole, J L; Molinari, J A

    1976-01-01

    The present investigation examined the specificity of the salivary immune response of axenic and conventional mice to topically administered Salmonella typhi, S. gallinarum, and S. typhimurium. Specific antibacterial antibodies were determined by passive hemagglutination and bacterial agglutination. Reciprocal antibody titers up to 320 were detected in saliva from mice immunized and assayed with homologous antigens. Antibodies to heterologous immunogens exhibited lower mean titers of 10 to 20 under identical conditions. High concentrations of specific antibodies to the somatic (O) antigen were detected in the saliva of mice administered these microorganisms; however, no significant differences in serum antibody levels were detectable after oral immunization. Only low levels of specific antiflagellar (H) antibodies were demonstrated in the saliva of immunized mice, whereas mean reciprocal titers of 20 were observed in the serum. Antibodies to the Vi antigen of S. typhi were detected in the saliva and serum of only those mice administered formalin-treated S. typhi. Examination of the classes of antibody elicited by these organisms indicated that immunoglobulin A (IgA) was the predominant class in saliva against the O antigens. The salivary response to the H antigens was comprised of both IgG and IgA, whereas the specific serum immunoglobulins were consistent with a primary humoral immune reaction. Local antibodies formed in response to the Vi antigen were exclusively IgG. Serum immunoglobulins produced after peroral administration of the somatic and virulence antigens were limited to the IgM class. PMID:765284

  13. [Managing patients with therapeutic antibodies in odontostomatology].

    PubMed

    Demoersman, J; Soueidan, A; Corre, P; Pers, J O

    2014-06-01

    Immunotherapies, particularly therapeutic antibodies, are increasingly used in the treatment of many autoimmune or oncological diseases. Patients treated with therapeutic antibodies may present with an increased risk of infection or of osteonecrosis of the jaws (ONJ). There is currently no consensus on the management of patients treated with therapeutic antibodies. These treatments are mainly used in hospitals, but they have been increasingly prescribed in ambulatory treatment for patients undergoing oral care. It is therefore important to establish therapeutic precautions for these patients. We had for aim to describe these antibody therapies, their indications, their potentially adverse effects in the oral cavity and to review the latest recommendations. PMID:24797731

  14. Preparation of astatine-labeled monoclonal antibodies

    SciTech Connect

    Milesz, S.; Norseev, Yu.V.; Szucs, Z. [Joint Inst. for Nuclear Research, Dubna (Russian Federation)]|[Central Inst. of Physical Research, Budapest (Hungary)

    1995-07-01

    In the cationic state astatine forms a stable complex with diethylenetriaminepentaacetic acid. Thanks to this complex, astatine can be bound to monoclonal antibodies of the RYa{sub 1} type. The most favorable conditions for preparing astatine-labeled antibodies are established. The chromatographic analysis and electromigration experiments showed that astatine is firmly linked to a biomolecule in vitro and it did not escape from labeled monoclonal antibodies even under treatment with such highly effective astatine-complexing agent as thiourea. The immune activity of astatine-labeled antibodies did not change even after 20 h.

  15. Plant cardiac glycosides and digoxin Fab antibody.

    PubMed

    Cheung, K; Urech, R; Taylor, L; Duffy, P; Radford, D

    1991-10-01

    The potential application of the Digoxin Fab antibody (Wellcome Digibind) in the clinical management of plant poisoning was investigated. The cardiac glycoside contents of various Australian plants were studied using immunoassay techniques. The cross-reactions of the Fab antibody and two digoxin assay antibodies against extracts of these plants were also studied. Results obtained indicated that the Digibind antibody cross-reacted with a wide range of glycosides contained in Australian plants and therefore could be of use in the treatment of life-threatening plant poisoning. PMID:1931226

  16. Monoclonal Antibody Therapy for Cancer

    Microsoft Academic Search

    Christoph Rader

    \\u000a Since the approval of rituximab (Rituxan®) for the treatment of B-cell non-Hodgkin’s lymphoma (B-NHL) in 1997, nine additional monoclonal antibodies (mAbs) have been\\u000a approved by the FDA for cancer therapy. Currently, more than 1,300 clinical studies registered at ClinicalTrials.gov investigate\\u000a mAb therapy of cancer, including more than 150 phase III clinical trials. In concert with their clinical acceptance, mAbs\\u000a in

  17. Function Blocking Antibodies to Neuropilin-1 Generated from a Designed Human Synthetic Antibody Phage Library

    Microsoft Academic Search

    Wei-Ching Liang; Mark S. Dennis; Scott Stawicki; Yvan Chanthery; Qi Pan; Yongmei Chen; Charles Eigenbrot; JianPing Yin; Alexander W. Koch; Xiumin Wu; Napoleone Ferrara; Anil Bagri; Marc Tessier-Lavigne; Ryan J. Watts; Yan Wu

    2007-01-01

    Non-immune (naďve) antibody phage libraries have become an important source of human antibodies. The synthetic phage antibody library described here utilizes a single human framework with a template containing human consensus complementarity-determining regions (CDRs). Diversity of the libraries was introduced at select CDR positions using tailored degenerate and trinucleotide codons that mimic natural human antibodies. Neuropilin-1 (NRP1), a cell-surface receptor

  18. Camel Single-domain Antibodies as Modular Building Units in Bispecific and Bivalent Antibody Constructs

    Microsoft Academic Search

    Katja Els Conrath; Mark Lauwereys; Lode Wyns; Serge Muyldermans

    2001-01-01

    Single-domain antibodies against various antigens are isolated from the unique heavy-chain antibodies of immunized camels and llamas. These minimal sized binders are very robust and bind the antigen with high affinity in a monomeric state. We evaluated the feasibil- ity to produce soluble, functional bispecific and bivalent antibodies in Escherichia coli with camel single-domain antibody fragments as building blocks. Two

  19. Transplacental transfer of immune antibodies in the mouse demonstrated by antibody labeled in vivo with tritium

    E-print Network

    McKinney, Hubert Eugene

    1971-01-01

    TRANSPLACENTAL TRANSFER OF IMMUNE ANTIBODIES IN THE MOUSE DEMONSTRATED BY ANTIBODY LABELED IN VIVO WITH TRITIUM A Thesis by HUBERT EUGENE MCKINNEY Submitted to the Graduate College of Texas ASM University in partial fulfillment... of the requirement for the degree of MASTER OF SCIENCE August 1971 Major Sub] ect: Laboratory Animal Medicine TRAYSPLACENTAL TRANSFER OF IMMUNE ANTIBODIES IN THE MOUSE DEMONSTRATED BY ANTIBODY LABELED IN VIVO WITH TRITIUM A Thesis by HUBERT EUGENE MCKINNEY...

  20. Engineered antibody fragments and the rise of single domains

    Microsoft Academic Search

    Philipp Holliger; Peter J Hudson

    2005-01-01

    With 18 monoclonal antibody (mAb) products currently on the market and more than 100 in clinical trials, it is clear that engineered antibodies have come of age as biopharmaceuticals. In fact, by 2008, engineered antibodies are predicted to account for >30% of all revenues in the biotechnology market. Smaller recombinant antibody fragments (for example, classic monovalent antibody fragments (Fab, scFv))

  1. Clinical and Therapeutic Aspects Associated to Phospholipid Binding Antibodies (Lupus Anticoagulant and Anticardiolipin Antibodies)

    Microsoft Academic Search

    J. Ordi-Ros; P. Pérez-Pemán; J. Monasterio

    1994-01-01

    Antiphospholipid antibodies (APA) comprise a family of immunoglobulins characterized by their pattern of reactivity in a number of laboratory tests. Included in this family are lupus anticoagulant (LA) anticardiolipin antibodies (ACA) and antibodies causing biologic false positive serologic tests for syphilis (BFP-STS). LA and ACA occur in a variety of conditions, including other autoimmunes disorders, infectious diseases, neoplasic disorders, in

  2. Antibodies Act Jointly to Promote Inflammation in Rheumatoid Arthritis

    MedlinePLUS

    ... ease the symptoms. Molecules called antibodies usually help combat infections by binding to foreign molecules like viral ... the antibodies to bind to their normal human targets, creating what are known as complexes. These antibody- ...

  3. Radiohalogenated half-antibodies and maleimide intermediate therefor

    DOEpatents

    Kassis, A.I.; Khawli, L.A.

    1991-02-19

    N-(m-radiohalophenyl) maleimide can be conjugated with a reduced antibody having a mercapto group to provide a radiolabeled half-antibody having immunological specific binding characteristics of whole antibody. No Drawings

  4. Bioconjugation of antibodies to horseradish peroxidase (hrp)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The bioconjugation of an antibody to an enzymatic reporter such as horseradish peroxidase (HRP) affords an effective mechanism by which immunoassay detection of a target antigen can be achieved. The use of heterobifunctional cross—linkers to covalently link antibodies to HRP provides a simple and c...

  5. Complement in antibody-based tumor therapy.

    PubMed

    Derer, Stefanie; Beurskens, Frank J; Rosner, Thies; Peipp, Matthias; Valerius, Thomas

    2014-01-01

    Monoclonal antibodies constitute a major treatment option for many tumor patients. Due to their specific recognition sites in their constant Fc regions, antibodies are able to trigger antibody-dependent cell-mediated cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC). While the contribution of ADCC to clinical efficacy has been strengthened by observations that patients with favorable Fc? receptor polymorphisms display better response rates to therapeutic antibodies, the contribution of CDC to their clinical efficacy remains controversial. In the background of high expression of complement-regulatory proteins on tumor cells as well as of the fact that some therapeutic antibodies lack the capacity to trigger efficient CDC, strategies have been implemented to improve either the capacity of antibodies to initiate the complement cascade or to interfere with tumor cells' resistance mechanisms. Although both strategies have demonstrated therapeutic benefit in vitro and in murine models, CDC-enhanced antibodies-to the best of our knowledge-have not been clinically tested, and evidence for the potential of CDC-optimizing approaches has yet to be generated in humans. Hence, the potency of complement activation and its impact on the clinical efficacy of therapeutic antibodies still remains to be elucidated in clinical trials encompassing novel complement-enhancing molecules. PMID:24941073

  6. Induction and detection of antibodies to squalene

    Microsoft Academic Search

    Gary R Matyas; Mangala Rao; Carl R Alving

    2002-01-01

    An improved high throughput assay for measuring murine antibodies to squalene (SQE) is described. The assay is highly reproducible and sensitive and can detect 80 ng\\/ml of antibody to SQE. The assay, an ELISA, is similar to our previously described assay in which plates containing PVDF membranes were used [J. Immunol. Methods 245 (2000) 1]. The PVDF plates worked well

  7. Structure and specificity of lamprey monoclonal antibodies

    E-print Network

    Ronquist, Fredrik

    anthracis spores. The recombinant VLR-B antibodies possess 8­10 uniform subunits that collectively bind describe the production of recombinant VLR-B antibodies specific for BclA, a major coat protein of Bacillus abortus, and human red blood cells (6­10). More recently, we demonstrated that immunization with Bacillus

  8. Original article Production of monoclonal antibodies against

    E-print Network

    Paris-Sud XI, Université de

    Original article Production of monoclonal antibodies against equine influenza : application 1988) Summary ― Monoclonal antibodies (Mo Abs) were prepared against influenza/A/equine/Prague/1. These monoclonals were tested against the 2 reference strains, 8 field strains of equine influenza virus, 3 human

  9. How to successfully patent therapeutic antibodies.

    PubMed

    Lahrtz, Fritz

    2015-04-01

    Therapeutic antibodies have become an established class of drugs for the treatment of a variety of diseases, especially cancer and autoimmune/inflammatory disorders, and a sufficient patent protection is a prerequisite for their successful commercialization. As monoclonal antibodies and their therapeutic potential have been well known for decades, the mere production of yet another therapeutic antibody is in many jurisdictions not considered a patentable invention. In contrast, antibodies with novel structural features and/or improved properties may be patentable. When drafting the claims, care should be taken to obtain a broad patent scope that protects both the antibody of interest and related antibodies having the same functional features, thereby preventing competitors from marketing a functionally equivalent antibody. Furthermore, the application should contain experimental evidence showing the improved properties of the claimed antibody. After the filing of a priority patent application, patent protection should be initiated at least in countries that are of particular commercial importance. Subsequent inventions relating to novel uses, formulations, dosage regimens, and combinations with other treatment modalities should be protected by further patent applications to extend patent term. PMID:25614506

  10. Monoclonal antibodies specific for mercuric ions.

    PubMed Central

    Wylie, D E; Lu, D; Carlson, L D; Carlson, R; Babacan, K F; Schuster, S M; Wagner, F W

    1992-01-01

    Monoclonal antibodies (mAbs) that react with soluble mercuric ions have been produced by injection of BALB/c mice with a hapten-carrier complex designed to maximize exposure of the metal to the immune system. Three hybridomas producing antibodies that reacted with bovine serum albumin (BSA)-glutathione-HgCl, but not with BSA-glutathione, were isolated from the spleen of a mouse given multiple injections with glutathione-HgCl conjugated to keyhole limpet hemocyanin. Stable subclones were established from two of these antibodies, designated mAb 4A10 and mAb 1F10. The binding of both antibodies to immobilized BSA-glutathione-HgCl was inhibited by soluble HgCl2, and dissociation constants for mercuric chloride binding were 2.3 and 3.7 nM for mAbs 4A10 and 1F10, respectively. Both antibodies bound mercuric acetate with similar affinities, demonstrating that the antibodies were capable of binding to mercuric ions in the presence of a different counterion than the one used in the immunogen. Reactions were not observed with other metal cations by either antibody. These data demonstrate the successful induction of antibodies that react very specifically with mercuric ions in solution regardless of the presence of a carrier. PMID:1570337

  11. Cytolytic Antibodies to Melanocytes in Vitiligo

    Microsoft Academic Search

    Jian Cui; Yuko Arita; Jean-Claude Bystryn

    1993-01-01

    Patients with vitiligo have been found to have circulating antibodies to pigment cells. To evaluate the functional activity of these antibodies, a highly sensitive europium release assay was used to compare complement-mediated cytolysis of human melanocytes by sera of 56 patients with vitiligo (20 with active disease, 25 with inactive disease, 11 with unidentified disease activity) and 47 control individuals.

  12. DNA Repair, Antibody Diversity, and Aging

    Microsoft Academic Search

    R. C. Johnson; A. C. Wang

    1985-01-01

    Proposed relationships of DNA repair, mutation, and the process of generation of antibody diversity allow new insights into the mechanism of aging. Pathways of antibody development are reviewed with special attention to steps which generate diversity. The normal process of combinatorial fusion of V region gene segments (i.e. V, D, and J) coding for the entire V region, plus the

  13. Synthetic Antibodies for Reversible Cell Recognition

    NASA Astrophysics Data System (ADS)

    Zhou, Jing Zhou

    2011-12-01

    Antibody-mediated cell recognition plays a critical role in various biological and biomedical applications. However, strong antibody-cell interactions can lead to the difficulty of separating antibodies from the bound cells in a simple and non-destructive manner, which is often necessary to numerous applications such as cell sorting or separation. Thus, this thesis research is aimed to create an antibody-like nanomaterial with the function of reversible cell recognition It was hypothesized that nucleic acid aptamer and dendrimer could be used as fundamental structural components to develop an antibody-like nanomaterial. The aptamer functions as the binding site of an antibody; the dendrimer is used as a robust, defined nano-scaffold to support the aptamer and to carry small molecules (e.g., fluorophores). To test this hypothesis, a novel method was first developed to discover the essential nucleotides of full-length aptamers to mimic the binding sites of antibodies. The essential nucleotides were further conjugated with a dendrimer to synthesize a monovalent aptamer-dendrimer nanomaterial. The results clearly showed that the essential nucleotides could maintain high affinity and specificity after tethered on dendrimer surface. To further test the hypothesis that antibody-like nanomaterials can be rationally designed to acquire the capability of reversible cell recognition, an aptamer that was selected at 0 °C was used as a model to synthesize a "Y-shaped" nanomaterial by conjugating two aptamers to the same dendrimer. The results showed that the nanomaterial-cell interaction could be affected by the distance between two binding aptamers. In addition, the "Y-shaped" antibody-like nanomaterial could bind target cells more strongly than its monovalent control. Importantly, the strong cell-nanomaterial interaction could be rapidly reversed when the temperature was shifted from 0 °C to 37 °C. In summary, we developed a synthetic antibody that can not only mimic the functional structure and cell recognition capability of antibodies, but also possess specific features that natural antibodies do not possess. This study has opened a new avenue for developing synthetic antibodies and has also advanced the understanding of the functionality of multivalent nanomaterials. This novel synthetic antibody holds great potential for various biological and biomedical applications such as cell separation.

  14. Clearance of pathological antibodies using biomimetic nanoparticles

    PubMed Central

    Copp, Jonathan A.; Fang, Ronnie H.; Luk, Brian T.; Hu, Che-Ming J.; Gao, Weiwei; Zhang, Kang; Zhang, Liangfang

    2014-01-01

    Pathological antibodies have been demonstrated to play a key role in type II immune hypersensitivity reactions, resulting in the destruction of healthy tissues and leading to considerable morbidity for the patient. Unfortunately, current treatments present significant iatrogenic risk while still falling short for many patients in achieving clinical remission. In the present work, we explored the capability of target cell membrane-coated nanoparticles to abrogate the effect of pathological antibodies in an effort to minimize disease burden, without the need for drug-based immune suppression. Inspired by antibody-driven pathology, we used intact RBC membranes stabilized by biodegradable polymeric nanoparticle cores to serve as an alternative target for pathological antibodies in an antibody-induced anemia disease model. Through both in vitro and in vivo studies, we demonstrated efficacy of RBC membrane-cloaked nanoparticles to bind and neutralize anti-RBC polyclonal IgG effectively, and thus preserve circulating RBCs. PMID:25197051

  15. Canadian Public Health Laboratory Network laboratory guidelines for the use of direct tests to detect syphilis in Canada

    PubMed Central

    Tsang, Raymond SW; Morshed, Muhammad; Chernesky, Max A; Jayaraman, Gayatri C; Kadkhoda, Kamran

    2015-01-01

    Treponema pallidum subsp. pallidum and/or its nucleic acid can be detected by various methods such as microscopy, rabbit infectivity test or polymerase chain reaction (PCR) tests. The rabbit infectivity test for T. pallidum, although very sensitive, has been discontinued from most laboratories due to ethical issues related to the need for animal inoculation with live T. pallidum, the technically demanding procedure and long turnaround time for results, thus making it impractical for routine diagnostic use. Dark-field and phase-contrast microscopy are still useful at clinic- or hospital-based laboratories for near-bedside detection of T. pallidum in genital, skin or mucous lesions although their availability is decreasing. The lack of reliable and specific anti-T. pallidum antibodies and its inferior sensitivity to PCR may explain why the direct fluorescent antibody test for T. pallidum is not widely available for clinical use. Immunohistochemical staining for T. pallidum also depends on the availability of specific antibodies, and the method is only applicable for histopathological examination of biopsy and autopsy specimens necessitating an invasive specimen collection approach. With recent advances in molecular diagnostics, PCR is considered to be the most reliable, versatile and practical for laboratories to implement. In addition to being an objective and sensitive test for direct detection of Treponema pallidum subsp. pallidum DNA in skin and mucous membrane lesions, the resulting PCR amplicons from selected gene targets can be further characterized for antimicrobial (macrolide) susceptibility testing, strain typing and identification of T. pallidum subspecies. PMID:25798160

  16. 9 CFR 113.452 - Erysipelothrix Rhusiopathiae Antibody.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE VIRUSES, SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS STANDARD REQUIREMENTS Antibody Products § 113.452 Erysipelothrix Rhusiopathiae Antibody. Erysipelothrix...

  17. 9 CFR 113.452 - Erysipelothrix Rhusiopathiae Antibody.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ...PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE VIRUSES, SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS STANDARD REQUIREMENTS Antibody Products § 113.452 Erysipelothrix Rhusiopathiae Antibody. Erysipelothrix...

  18. 9 CFR 113.452 - Erysipelothrix Rhusiopathiae Antibody.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ...PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE VIRUSES, SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS STANDARD REQUIREMENTS Antibody Products § 113.452 Erysipelothrix Rhusiopathiae Antibody. Erysipelothrix...

  19. Monoclonal Antibody Therapies against Anthrax

    PubMed Central

    Chen, Zhaochun; Moayeri, Mahtab; Purcell, Robert

    2011-01-01

    Anthrax is a highly lethal infectious disease caused by the spore-forming bacterium Bacillus anthracis. It not only causes natural infection in humans but also poses a great threat as an emerging bioterror agent. The lethality of anthrax is primarily attributed to the two major virulence factors: toxins and capsule. An extensive effort has been made to generate therapeutically useful monoclonal antibodies to each of the virulence components: protective antigen (PA), lethal factor (LF) and edema factor (EF), and the capsule of B. anthracis. This review summarizes the current status of anti-anthrax mAb development and argues for the potential therapeutic advantage of a cocktail of mAbs that recognize different epitopes or different virulence factors. PMID:22069754

  20. ANTIBODIES TO INTESTINAL MICROVILLOUS MEMBRANES

    PubMed Central

    Kopp, William L.; Trier, Jerry S.; Mackenzie, Iain L.; Donaldson, Robert M.

    1968-01-01

    Antibodies (AbMVM) were produced in rabbits to microvillous membranes isolated from hamster small bowel. Incubation of frozen sections of hamster small bowel with fluorescein-labeled AbMVM showed specific reaction with brush borders, but not with other intestinal cellular components. Electron microscopy with ferritin-conjugated AbMVM localized the antigens more precisely to the surface mucopolysaccharide coat of the brush borders. AbMVM also reacted with the brush border of colon and of proximal renal tubules of hamsters but not with those of hamster stomach or gall bladder. It also reacted with the brush borders of some rat and human tissues, but not with those of rabbits. In addition, fluorescent-labeled AbMVM combined specifically with cell walls of some yeasts, but not of several bacteria. AbMVM also contained a weak precipitin to a component of hamster serum, which migrated like prealbumin in immunoelectrophoresis. PMID:4969881

  1. Monoclonal antibody therapies against anthrax.

    PubMed

    Chen, Zhaochun; Moayeri, Mahtab; Purcell, Robert

    2011-08-01

    Anthrax is a highly lethal infectious disease caused by the spore-forming bacterium Bacillus anthracis. It not only causes natural infection in humans but also poses a great threat as an emerging bioterror agent. The lethality of anthrax is primarily attributed to the two major virulence factors: toxins and capsule. An extensive effort has been made to generate therapeutically useful monoclonal antibodies to each of the virulence components: protective antigen (PA), lethal factor (LF) and edema factor (EF), and the capsule of B. anthracis. This review summarizes the current status of anti-anthrax mAb development and argues for the potential therapeutic advantage of a cocktail of mAbs that recognize different epitopes or different virulence factors. PMID:22069754

  2. HIV antibody detection in oral fluids.

    PubMed

    1993-01-01

    Oral fluids are a mixture of saliva and oral mucosal transudates (OMT). Saliva is a product of the salivary glands and contains mostly IgA, while OMT is mostly fluid in the subgingival space derived from the passive transport of plasma and contains mostly IgG. The IgG concentration, however, is much lower than that in serum. Testing for antibodies to HIV in oral fluids has been proposed as an alternative to antibody testing in blood. The fluids may be collected directly by dribbling into a receptacle and via absorption onto pads using specially designed collection devices. Only one commercially available HIV antibody test is, however, specifically designed for use with oral fluid samples. Some existing commercial tests designed to detect antibody to HIV in blood samples have been modified for use with oral fluids, but only limited information is available on their performance. Several studies suggest that using tests to detect HIV antibodies in oral fluids may be adequate for some situations, but a number of issues remain to be addressed. WHO therefore recommends that a full evaluation of the detection of HIV antibody in oral fluids be undertaken to address the issues before recommendations on HIV antibody testing using oral fluids are made. Such evaluation would gather information on the accuracy and cost-effectiveness of testing oral fluids. Ethical considerations when performing HIV testing using oral fluids would be the same as those for blood: non-coercion, informed consent, counseling, and confidentiality. PMID:8261568

  3. Structure Based Antibody-Like Peptidomimetics

    PubMed Central

    Murali, Ramachandran; Greene, Mark I.

    2012-01-01

    Biologics such as monoclonal antibodies (mAb) and soluble receptors represent new classes of therapeutic agents for treatment of several diseases. High affinity and high specificity biologics can be utilized for variety of clinical purposes. Monoclonal antibodies have been used as diagnostic agents when coupled with radionuclide, immune modulatory agents or in the treatment of cancers. Among other limitations of using large molecules for therapy the actual cost of biologics has become an issue. There is an effort among chemists and biologists to reduce the size of biologics which includes monoclonal antibodies and receptors without a reduction of biological efficacy. Single chain antibody, camel antibodies, Fv fragments are examples of this type of deconstructive process. Small high-affinity peptides have been identified using phage screening. Our laboratory used a structure-based approach to develop small-size peptidomimetics from the three-dimensional structure of proteins with immunoglobulin folds as exemplified by CD4 and antibodies. Peptides derived either from the receptor or their cognate ligand mimics the functions of the parental macromolecule. These constrained peptides not only provide a platform for developing small molecule drugs, but also provide insight into the atomic features of protein-protein interactions. A general overview of the reduction of monoclonal antibodies to small exocyclic peptide and its prospects as a useful diagnostic and as a drug in the treatment of cancer are discussed. PMID:24288089

  4. Progress towards recombinant anti-infective antibodies

    PubMed Central

    Pai, Jennifer C.; Sutherland, Jamie N.; Maynard, Jennifer A.

    2009-01-01

    The global market for monoclonal antibody therapeutics reached a total of $11.2 billion in 2004, with an impressive 42% growth rate over the previous five years and is expected to reach ~$34 billion by 2010. Coupled with this growth are stream-lined product development, production scale-up and regulatory approval processes for the highly conserved antibody structure. While only one of the 21 current FDA-approved antibodies, and one of the 38 products in advanced clinical trials target infectious diseases, there is increasing academic, government and commercial interest in this area. Synagis, an antibody neutralizing respiratory syncitial virus (RSV), garnered impressive sales of $1.1 billion in 2006 in spite of its high cost and undocumented effects on viral titres in human patients. The success of anti-RSV passive immunization has motivated the continued development of anti-infectives to treat a number of other infectious diseases, including those mediated by viruses, toxins and bacterial/fungal cells. Concurrently, advances in antibody technology suggest that cocktails of several monoclonal antibodies with unique epitope specificity or single monoclonal antibodies with broad serotype specificity may be the most successful format. Recent patents and patent applications in these areas will be discussed as predictors of future anti-infective therapeutics. PMID:19149692

  5. Antibodies Against Three Forms of Urokinase

    NASA Technical Reports Server (NTRS)

    Morrison, Dennis R.; Atassi, M. Zouhair

    2007-01-01

    Antibodies that bind to preselected regions of the urokinase molecule have been developed. These antibodies can be used to measure small quantities of each of three molecular forms of urokinase that could be contained in microsamples or conditioned media harvested from cultures of mammalian cells. Previously available antibodies and assay techniques do not yield both clear distinctions among, and measurements of, all three forms. Urokinase is a zymogen that is synthesized in a single-chain form, called ScuPA, which is composed of 411 amino acid residues (see figure). ScuPA has very little enzyme activity, but it can be activated in two ways: (1) by cleavage of the peptide bond lysine 158/isoleucine 159 and the loss of lysine 158 to obtain the high molecular-weight (HMW) form of the enzyme or (2) by cleavage of the bond lysine 135/lysine 136 to obtain the low-molecular-weight (LMW) form of the enzyme. The antibodies in question were produced in mice and rabbits by use of peptides as immunogens. The peptides were selected to obtain antibodies that bind to regions of ScuPA that include the lysine 158/isoleucine 159 and the lysine 135/lysine 136 bonds. The antibodies include monoclonal and polyclonal ones that yield indications as to whether either of these bonds is intact. The polyclonal antibodies include ones that preferentially bind to the HMW or LMW forms of the urokinase molecule. The monoclonal antibodies include ones that discriminate between the ScuPA and the HMW form. A combination of these molecular-specific antibodies will enable simultaneous assays of the ScuPA, HMW, and LMW forms in the same specimen of culture medium.

  6. SAbDab: the structural antibody database

    PubMed Central

    Dunbar, James; Krawczyk, Konrad; Leem, Jinwoo; Baker, Terry; Fuchs, Angelika; Georges, Guy; Shi, Jiye; Deane, Charlotte M.

    2014-01-01

    Structural antibody database (SAbDab; http://opig.stats.ox.ac.uk/webapps/sabdab) is an online resource containing all the publicly available antibody structures annotated and presented in a consistent fashion. The data are annotated with several properties including experimental information, gene details, correct heavy and light chain pairings, antigen details and, where available, antibody–antigen binding affinity. The user can select structures, according to these attributes as well as structural properties such as complementarity determining region loop conformation and variable domain orientation. Individual structures, datasets and the complete database can be downloaded. PMID:24214988

  7. Fc engineering of antibodies and antibody derivatives by primary sequence alteration and their functional characterization.

    PubMed

    Derer, Stefanie; Kellner, Christian; Rösner, Thies; Klausz, Katja; Glorius, Pia; Valerius, Thomas; Peipp, Matthias

    2014-01-01

    Therapeutic antibodies used in the treatment of cancer patients are able to mediate diverse effector mechanisms. Dependent on tumor entity, localization, and tumor burden different effector mechanisms may contribute to the in vivo antitumor activity to a variable degree. Especially Fc-mediated effector functions such as antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) have been suggested as being important for the in vivo activity of therapeutic antibodies like rituximab or trastuzumab. In recent years, several strategies have been pursued to further optimize the cytotoxic potential of monoclonal antibodies by modifying their Fc part (Fc engineering) with the ultimate goal to enhance antibody therapy.Since Fc engineering approaches are applicable to any Fc-containing molecule, strategies to enhance CDC or ADCC activity of full antibodies or scFv-Fc fusion proteins by altering the primary Fc sequence are described. PMID:24515488

  8. Antibody discovery: the use of transgenic mice to generate human monoclonal antibodies for therapeutics.

    PubMed

    Kellermann, Sirid Aimée; Green, Larry L

    2002-12-01

    Technical advances made in the 1980s and early 1990s resulted in monoclonal antibodies that are now approved for human therapy. Novel transgenic mouse strains provide a powerful technology platform for creating fully human monoclonal antibodies as therapeutics; ten such antibodies have entered clinical trials since 1998 and more are in preclinical testing. Improved transgenic mouse strains provide a powerful technology platform for creating human therapeutics in the future. PMID:12482519

  9. Production of fully human antibodies by transgenic mice

    Microsoft Academic Search

    Aya Jakobovits

    1995-01-01

    The ability to produce a diverse repertoire of fully human monoclonal antibodies may have significant applications to human therapy. One of the most promising approaches to the production of therapeutic human monoclonal antibodies is the creation of a mouse strain engineered to produce a large repertoire of human antibodies in the absence of mouse antibodies. Recently, such mice have been

  10. Avian Diagnostic and Therapeutic Antibodies to Viral Emerging Pathogens

    SciTech Connect

    David Bradley

    2011-03-31

    During the current period the following key objectives were achieved: demonstration of high titer antibody production by geese following immunization with inactived H1N1 virus; completion of the epitope mapping of West Nile Virus-specific goose antibodies and initiation of epitope mapping of H1N1 flu-specific goose antibodies; advancement in scalable purification of goose antibodies.

  11. Deciphering the Antibodyome - Peptide Arrays for Serum Antibody Biomarker Diagnostics

    Microsoft Academic Search

    Heiko Andresen; Carsten Grotzinger

    2009-01-01

    The analysis of antibodies in human serum is an established technique in the laboratory diagnosis of infectious as well as autoimmune diseases. The multitude of antibody reactions towards pathogens and likewise the antibody profile in autoimmune diseases does contain a wealth of proteomic (antibody) data that may constitute valuable diagnostic infor- mation with relevance for the patient's prognosis and response

  12. Myopathy with anti-HMGCR antibodies

    PubMed Central

    Alshehri, Ali; Choksi, Rati; Bucelli, Robert

    2015-01-01

    Objective: To analyze clinical features and myopathology changes in muscle fibers, connective tissue, and vessels in 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR) antibody–associated myopathies. Methods: Retrospective review of records and myopathologic features of 49 consecutive patients with myopathies and serum HMGCR antibodies. Results: Clinical features included onset age from 12 to 83 years, female predominance (67%), proximal, symmetric weakness (84%), muscle discomfort (78%), dysphagia (35%), systemic features, including skin rash and interstitial lung disease (37%), statin use (38%), and a high serum creatine kinase (83%). Myopathology included muscle fiber necrosis or regeneration (66%), myonuclear pathology (43%), perimysial connective tissue damage (61%), and lymphocytic foci (27%). Conclusions: Patients with HMGCR antibody–associated myopathies present with weakness and muscle discomfort and often have damage to both perimysial connective tissue and muscle fibers, with necrosis and myonuclear pathology. Only a minority of patients with HMGCR antibody–associated myopathies have a history of statin exposure.

  13. Localization of tumors by radiolabelled antibodies

    Microsoft Academic Search

    H. J. Hansen; F. J. Primus

    1975-01-01

    A method of utilizing radiolabelled antibodies to carcinoembryonic antigens for determining the site of tumors which produce or are associated with carcinoembryonic antigen is disclosed. 3 claims, no drawings.

  14. Antibodies against the calcium-binding protein

    SciTech Connect

    Chou, Mei; Jensen, K.G.; Sjolund, R.D. (Univ. of Iowa, Iowa City (USA)); Krause, K.H.; Campbell, K.P. (Univ. of Iowa College of Medicine, Iowa City (USA))

    1989-12-01

    Plant microsomes contain a protein clearly related to a calcium-binding protein, calsequestrin, originally found in the sarcoplasmic reticulum of muscle cells, responsible for the rapid release and uptake of Ca{sup 2+} within the cells. The location and role of calsequestrin in plant cells is unknown. To generate monoclonal antibodies specific to plant calsequestrin, mice were immunized with a microsomal fraction from cultured cells of Streptanthus tortuosus (Brassicaceae). Two clones cross-reacted with one protein band with a molecular weight equal to that of calsequestrin (57 kilodaltons) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. This band is able to bind {sup 45}Ca{sup 2+} and can be recognized by a polyclonal antibody against the canine cardiac muscle calsequestrin. Rabbit skeletal muscle calsequestrin cross-reacted with the plant monoclonal antibodies. The plant monoclonal antibodies generated here are specific to calsequestrin protein.

  15. Polynucleotides encoding anti-sulfotyrosine antibodies

    DOEpatents

    Bertozzi, Carolyn R. (Berkeley, CA); Kehoe, John (Saint Davids, PA); Bradbury, Andrew M. (Santa Fe, NM)

    2011-01-11

    The invention provides anti-sulfotyrosine specific antibodies capable of detecting and isolating polypeptides that are tyrosine-sulfated. The sulfotyrosine antibodies and antibody fragments of the invention may be used to discriminate between the non-sulfated and sulfated forms of such proteins, using any number of immunological assays, such ELISAs, immunoblots, Western Blots, immunoprecipitations, and the like. Using a phage-display system, single chain antibodies (scFvs) were generated and screened against tyrosine-sulfated synthetic peptide antigens, resulting in the isolation of scFvs that specifically recognize sulfotyrosine-containing peptides and/or demonstrate sulfotyrosine-specific binding in tyrosine sulfated proteins. The VH and VL genes from one such sulfotyrosine-specific scFv were employed to generate a full length, sulfotyrosine-specific immunoglobulin.

  16. Broadly neutralizing antibodies against influenza viruses

    PubMed Central

    Laursen, Nick S.; Wilson, Ian A.

    2014-01-01

    Despite available antivirals and vaccines, influenza infections continue to be a major cause of mortality worldwide. Vaccination generally induces an effective, but strain-specific antibody response. As the virus continually evolves, new vaccines have to be administered almost annually when a novel strain becomes dominant. Furthermore, the sporadic emerging resistance to neuraminidase inhibitors among circulating strains suggests an urgent need for new therapeutic agents. Recently, several cross-reactive antibodies have been described, which neutralize an unprecedented spectrum of influenza viruses. These broadly neutralizing antibodies generally target conserved functional regions on the major influenza surface glycoprotein hemagglutinin (HA). The characterization of their neutralization breadth and epitopes on HA could stimulate the development of new antibody-based antivirals and broader influenza vaccines. PMID:23583287

  17. Brain-Reactive Antibodies and Disease

    PubMed Central

    Diamond, B.; Honig, G.; Mader, S.; Brimberg, L.; Volpe, B.T.

    2015-01-01

    Autoimmune diseases currently affect 5–7% of the world's population; in most diseases there are circulating autoantibodies. Brain-reactive antibodies are present in approximately 2–3% of the general population but do not usually contribute to brain pathology. These antibodies penetrate brain tissue only early in development or under pathologic conditions. This restriction on their pathogenicity and the lack of correlation between serum titers and brain pathology have, no doubt, contributed to a delayed appreciation of the contribution of autoantibodies in diseases of the central nervous system. Nonetheless, it is increasingly clear that antibodies can cause damage in the brain and likely initiate or aggravate multiple neurologic conditions; brain-reactive antibodies contribute to symptomatology in autoimmune disease, infectious disease, and malignancy. PMID:23516983

  18. Antibody Fc: Linking Adaptive and Innate Immunity

    PubMed Central

    Reichert, Janice M.

    2014-01-01

    Antibody Fc: Linking Adaptive and Innate Immunity, edited by Margaret E. Ackerman and Falk Nimmerjahn and published by Academic Press, provides a highly detailed examination of the involvement of the antibody Fc in mechanisms critical to both innate and adaptive immune responses. Despite a recent increase in format diversity, most marketed antibodies are full-length IgG molecules and the majority of the commercial clinical pipeline of antibody therapeutics is composed of Fc-containing IgG molecules, which underscores the importance of understanding how the Fc domain affects biological responses. The book is divided into six sections that include a total of 20 chapters. In order of their appearance, the sections provide extensive coverage of effector mechanisms, effector cells, Fc receptors, variability of the Fc domain, genetic associations, and evolving areas.

  19. Antiphospholipid antibodies in critically ill patients

    PubMed Central

    Vassalo, Juliana; Spector, Nelson; de Meis, Ernesto; Soares, Márcio; Salluh, Jorge Ibrain Figueira

    2014-01-01

    Antiphospholipid antibodies are responsible for a wide spectrum of clinical manifestations. Venous, arterial and microvascular thrombosis and severe catastrophic cases account for a large morbidly/mortality. Through the connection between the immune, inflammatory and hemostatic systems, it is possible that these antibodies may contribute to the development of organ dysfunction and are associated with poor short and long-term prognoses in critically ill patients. We performed a search of the PubMed/MedLine database for articles written during the period from January 2000 to February 2013 to evaluate the frequency of antiphospholipid antibodies in critically ill patients and their impact on the outcomes of these patients. Only eight original studies involving critically ill patients were found. However, the development of antiphospholipid antibodies in critically ill patients seems to be frequent, but more studies are necessary to clarify their pathogenic role and implications for clinical practice. PMID:25028953

  20. Antibodies to watch in 2010

    PubMed Central

    2010-01-01

    Monoclonal antibodies (mAbs) are a burgeoning class of therapeutics, with more than 25 approved in countries worldwide. Novel molecules are entering clinical study at a rate of nearly 40 per year, and the commercial pipeline includes approximately 240 mAb therapeutics in clinical studies that have not yet progressed to regulatory approval or been approved. Of particular interest are the 26 mAbs that are currently at Phase 3, when safety and efficacy data critical to approval is established. Phase 3 study lengths are typically two to four years, so results for some studies might be announced in 2010, but data from others might not be presented until 2014. This overview of the 26 candidates provides a brief description of the background and the on-going Phase 3 studies of each mAb. Additional mAbs that have progressed to regulatory review or been approved may also be in Phase 3 studies, but these, as well as Fc fusion proteins, have been excluded. Due to the large body of primary literature about the 26 candidates, only selected references are given, with a focus on recent publications and articles that were relevant to Phase 3 studies. Current as of October 2009, the results presented here will serve as a baseline against which future progress can be measured. PMID:20065640

  1. Lung adenocarcinoma and antiphospholipid antibodies.

    PubMed

    de Meis, Ernesto; Monteiro, Robson Q; Levy, Roger A

    2009-05-01

    Thrombosis is a frequent finding in cancer patients, being referred to as a poor prognostic factor. The mechanisms underlying the thrombophilic state in malignancy are not well elucidated but involve a complex interaction between tumor and host cells as well as the hemostatic system. A number of studies have demonstrated the presence of antiphospholipid antibodies (aPL) in cancer patients, suggesting a potential role in tumor-associated thrombosis. A prospective analysis has been performed in a group of lung adenocarcinoma patients in respect to the presence of aPL and thrombotic manifestations. Lupus anticoagulant (LAC) was identified in 61 out of 105 patients and it correlated highly with thrombosis (22/61, LAC positive vs 2/44, LAC negative RR=7.93; p<0.001). On the other hand, patients that displayed IgM anti-beta2-glycoprotein I (abeta2GPI) (22/80) showed an unexpected decrease in thrombosis risk (2/22, with IgM abeta2GPI vs 18/58, without IgM abeta2GPI RR=0.29; p=0.04). Considerations on the mechanisms that link cancer, thrombosis and aPL are discussed in this article. PMID:19185619

  2. [Anti-basal ganglia antibody].

    PubMed

    Hayashi, Masaharu

    2013-04-01

    Sydenham's chorea (SC) is a major manifestation of rheumatic fever, and the production of anti-basal ganglia antibodies (ABGA) has been proposed in SC. The pathogenesis is hypothesized as autoimmune targeting of the basal ganglia via molecular mimicry, triggered by streptococcal infection. The spectrum of diseases in which ABGA may be involved has been broadened to include other extrapyramidal movement disorders, such as tics, dystonia, and Parkinsonism, as well as other psychiatric disorders. The autoimmune hypothesis in the presence and absence of ABGA has been suggested in Tourette's syndrome (TS), early onset obsessive-compulsive disorders (OCD), and pediatric autoimmune neuropsychiatric disorders associated with streptococcal infections (PANDAS). Recently, the relationship between ABGA and dopamine neurons in the basal ganglia has been examined, and autoantibodies against dopamine receptors were detected in the sera from patients with basal ganglia encephalitis. In Japan, the occurrence of subacute encephalitis, where patients suffer from episodes of altered behavior and involuntary movements, has increased. Immune-modulating treatments are effective, indicating the involvement of an autoimmune mechanism. We aimed to detect the anti-neuronal autoantibodies in such encephalitis, using immunohistochemical assessment of patient sera. The sera from patients showing involuntary movements had immunoreactivity for basal ganglia neurons. Further epitopes for ABGA will be investigated in basal ganglia disorders other than SC, TS, OCD, and PANDAS. PMID:23568985

  3. Generation of novel recombinant antibodies against nitrotyrosine by antibody phage display.

    PubMed

    Hof, Danielle; Cooksley-Decasper, Seraina; Moergeli, Sandra; von Eckardstein, Arnold

    2011-01-01

    Nitrotyrosine is a posttranslational protein modification that occurs under oxidative and nitrosative stress, and plays an important role in numerous pathological conditions. To analyse nitrotyrosine formation several commercial monoclonal and polyclonal antibodies reacting with 3-nitrotyrosine have been developed which however do not work properly in all required assays. Here, antibody phage display was used to select recombinant antibodies that specifically react with nitrotyrosine in various protein contexts. Nine initial selections were carried out, using synthetic peptides, peroxynitrite-modified proteins and conjugated proteins as antigens. Four antibodies were isolated that each exhibited a characteristic binding reactivity that greatly depended on the antigens that were used for their selections. In general, the selections using small, synthetic and biotinylated peptides were the most successful approach. Subsequently, antibody 11B1 was affinity matured by error prone mutagenesis, resulting in the isolation of two antibodies, designated 47A7 and 47B1. Competition ELISA and immunoblotting after treatment with sodium dithionite further demonstrated the specificity of antibody 47B1 for nitrotyrosine. The results presented here demonstrate that antibody phage display is a useful method to isolate antibodies against posttranslational modifications, which are powerful tools in the proteomic era. PMID:21558620

  4. Antibody Affinity Purification to Detect Interacting Proteins

    Microsoft Academic Search

    Sonia Navarro; Lucio Comai

    \\u000a Affinity purification is a procedure based on the specific binding interactions between a ligand chemically coupled to a resin\\u000a and a target molecule. A common application is the use of antibody as immobilized ligands. The purification of antigens by\\u000a antibody-affinity chromatography is widely used to detect factors interacting with a protein of interest, and when coupled\\u000a to mass spectrometry, it

  5. Single domain camel antibodies: current status

    Microsoft Academic Search

    Serge Muyldermans

    2001-01-01

    The antigen-binding capacity of the paired variable domains of an antibody is well established. The observation that the isolated heavy chains of anti-hapten antibodies retain some antigen-binding capacity in the absence of light chains led to attempts to obtain an even smaller antigen-binding unit in a VH format. Unfortunately, the poor solubility, the reduced affinity for the antigen and the

  6. Engineering the variable region of therapeutic IgG antibodies

    PubMed Central

    Tsunoda, Hiroyuki; Kuramochi, Taichi; Sampei, Zenjiro; Ishii, Shinya; Hattori, Kunihiro

    2011-01-01

    Since the first generation of humanized IgG1 antibodies reached the market in the late 1990s, IgG antibody molecules have been extensively engineered. The success of antibody therapeutics has introduced severe competition in developing novel therapeutic monoclonal antibodies, especially for promising or clinically validated targets. Such competition has led researchers to generate so-called second or third generation antibodies with clinical differentiation utilizing various engineering and optimization technologies. Parent IgG antibodies can be engineered to have improved antigen binding properties, effector functions, pharmacokinetics, pharmaceutical properties and safety issues. Although the primary role of the antibody variable region is to bind to the antigen, it is also the main source of antibody diversity and its sequence affects various properties important for developing antibody therapeutics. Here we review recent research activity in variable region engineering to generate superior antibody therapeutics. PMID:21406966

  7. Monoclonal antibodies in acute lymphoblastic leukemia.

    PubMed

    Jabbour, Elias; O'Brien, Susan; Ravandi, Farhad; Kantarjian, Hagop

    2015-06-25

    With modern intensive combination polychemotherapy, the complete response (CR) rate in adults with acute lymphoblastic leukemia (ALL) is 80% to 90%, and the cure rate is 40% to 50%. Hence, there is a need to develop effective salvage therapies and combine novel agents with standard effective chemotherapy. ALL leukemic cells express several surface antigens amenable to target therapies, including CD20, CD22, and CD19. Monoclonal antibodies target these leukemic surface antigens selectively and minimize off-target toxicity. When added to frontline chemotherapy, rituximab, an antibody directed against CD20, increases cure rates of adults with Burkitt leukemia from 40% to 80% and those with pre-B ALL from 35% to 50%. Inotuzumab ozogamicin, a CD22 monoclonal antibody bound to calicheamicin, has resulted in marrow CR rates of 55% and a median survival of 6 to 7 months when given to patients with refractory-relapsed ALL. Blinatumomab, a biallelic T cell engaging the CD3-CD19 monoclonal antibody, also resulted in overall response rates of 40% to 50% and a median survival of 6.5 months in a similar refractory-relapsed population. Other promising monoclonal antibodies targeting CD20 (ofatumumab and obinutuzumab) or CD19 or CD20 and bound to different cytotoxins or immunotoxins are under development. Combined modalities of chemotherapy and the novel monoclonal antibodies are under investigation. PMID:25999456

  8. Anti Transglutaminase Antibodies Cause Ataxia in Mice

    PubMed Central

    Boscolo, Sabrina; Lorenzon, Andrea; Sblattero, Daniele; Florian, Fiorella; Stebel, Marco; Marzari, Roberto; Not, Tarcisio; Aeschlimann, Daniel; Ventura, Alessandro; Hadjivassiliou, Marios; Tongiorgi, Enrico

    2010-01-01

    Background Celiac disease (CD) is an autoimmune gastrointestinal disorder characterized by the presence of anti-transglutaminase 2 (TG2) and anti-gliadin antibodies. Amongst the neurological dysfunctions associated with CD, ataxia represents the most common one. Methods We analyzed by immunohistochemistry, the anti-neural reactivity of the serum from 20 CD patients. To determine the role of anti-TG2 antibodies in ataxia, two anti-TG2 single chain variable fragments (scFv), isolated from a phage-display IgA antibody library, were characterized by immunohistochemistry and ELISA, and injected in mice to study their effects on motor coordination. We found that 75% of the CD patient population without evidence of neurological involvement, has circulating anti-neural IgA and/or IgG antibodies. Two anti-TG2 scFvs, cloned from one CD patient, stained blood vessels but only one reacted with neurons. This anti-TG2 antibody showed cross reactivity with the transglutaminase isozymes TG3 and TG6. Intraventricular injection of the anti-TG2 or the anti-TG2/3/6 cross-reactive scFv provoked transient, equally intensive ataxia in mice. Conclusion The serum from CD patients contains anti-TG2, TG3 and TG6 antibodies that may potentially cause ataxia. PMID:20300628

  9. Decay of maternal antibodies in broiler chickens.

    PubMed

    Gharaibeh, Saad; Mahmoud, Kamel

    2013-09-01

    The objective of this study was to determine the decay rate of maternal antibodies against major broiler chicken pathogens. A total of 30 one-day-old broiler chicks were obtained from a commercial hatchery and reared in isolation. These chicks were retrieved from a parent flock that received a routine vaccination program. Chicks were bled at hatch and sequentially thereafter every 5 d through 30 d of age. Maternal antibody titers were measured by ELISA for avian encephalomyelitis (AEV), avian influenza virus (AIV), chicken anemia virus (CAV), infectious bursal disease virus (IBDV), infectious bronchitis virus (IBV), infectious laryngotracheitis virus (ILTV), Mycoplasma gallisepticum (MG), Mycoplasma synoviae (MS), and reovirus (Reo). Maternal antibody titers for Newcastle disease virus (NDV) were measured using a hemagglutination inhibition test. Half-life estimates of maternal antibody titers were 5.3, 4.2, 7, 5.1, 3.9, 3.8, 4.9, 4.1, 6.3, and 4.7 d for AEV, AIV, CAV, IBDV, IBV, ILTV, MG, MS, NDV, and Reo, respectively. The statistical analysis revealed significant differences among half-lives of maternal antibody titers against certain pathogens. Furthermore, all maternal antibody titers were depleted by 10 d of age except for IBDV. PMID:23960115

  10. Fourth World Antibody-Drug Conjugate Summit

    PubMed Central

    Beck, Alain; Lambert, John; Sun, Michael; Lin, Kedan

    2012-01-01

    The 4th World Antibody Drug Conjugate (WADC) Summit, organized by Hanson Wade was held on February 29?March 1, 2012 in Frankfurt, Germany, which was also the location for the Antibody Drug Conjugate Summit Europe held in February 2011. During the one year between these meetings, antibody drug conjugates (ADCs) have confirmed their technological maturity and their clinical efficacy in oncology. Brentuximab vedotin (ADCETRISTM) gained approval by the US Food and Drug Administration in August 2011 and trastuzumab emtansine (T-DM1) confirmed impressive clinical efficacy responses in a large cohort of breast cancer patients. During the 4th WADC meeting, antibody-maytansinoid conjugates were showcased by representatives of ImmunoGen (T-DM1, SAR3419, lorvotuzumab mertansine/IMGN801, IMGN529 and IMG853) and Biotest (BT-062). Data on antibody-auristatin conjugates were presented by scientists and clinicians from Seattle Genetics and Takeda (brentuximab vedotin), Pfizer (5T4-MMAF), Agensys/Astella (AGS-16M8F), Progenics (PSMA-ADC) and Genmab (anti-TF ADCs). Alternative payloads such as calicheamicins and duocarmycin used for preparation of ADCs were discussed by Pfizer and Synthon representatives, respectively. In addition, emerging technologies, including site-directed conjugation (Ambrx), a protein toxin as payload (Viventia), hapten-binding bispecific antibodies (Roche), and use of light activated drugs (Photobiotics), were also presented. Last but not least, progresses in solving Chemistry Manufacturing and Control, and pharmacokinetic issues were addressed by scientists from Genentech, Pfizer, Novartis and Pierre Fabre. PMID:22909934

  11. SPECIFIC FRACTIONATION OF HUMAN ANTIDEXTRAN ANTIBODIES

    PubMed Central

    Gelzer, Justus; Kabat, Elvin A.

    1964-01-01

    Human antidextran of one individual, absorbed specifically on sephadex, was fractionated into two populations of antibody molecules by successive elution with oligosaccharides of the isomaltose series of increasing size. The purified antibody fractions and some whole antidextran sera were found to fix complement with dextrans of molecular weight of 195,000 and above. It could be demonstrated by quantitative microcomplement fixation inhibition assays that the antibody eluted with isomaltotriose had a higher affinity for smaller oligosaccharides relative to isomaltohexaose, indicating a high content of antibody molecules with smaller combining sites, while with the second fraction, eluted with isomaltohexaose, the small haptens were very poor inhibitors and the larger oligosaccharides inhibited readily, presumably due to a higher proportion of molecules with larger combining site size. Assays of similarly prepared fractions, obtained from earlier bleedings of the same individual (1), with inhibition of complement fixation were in good agreement with those obtained by inhibition of precipitation. The two purified antidextran fractions were shown to differ with respect to their complement-fixing capacity. The fraction with molecules with smaller size-combining sites fixed only about half as much complement per unit antibody N as did the fraction containing largely molecules with larger combining sites suggesting that the strength of complement fixation is affected by the strength of the antigen-antibody interaction. PMID:14176295

  12. Antibodies and cancer therapy: versatile platforms for cancer immunotherapy

    PubMed Central

    Surana, Rishi; Wang, Shangzi

    2012-01-01

    Antibodies have emerged as important therapeutics for cancer. Recently, it has become clear that antibodies possess multiple clinically relevant mechanisms of action. Many clinically useful antibodies can manipulate tumour-related signalling. In addition, antibodies exhibit various immunomodulatory properties and, by directly activating or inhibiting molecules of the immune system, antibodies can promote the induction of anti-tumour immune responses. These immunomodulatory properties can form the basis for new cancer treatment strategies. PMID:20414205

  13. Thermodynamics of antibody-antigen interaction revealed by mutation analysis of antibody variable regions.

    PubMed

    Akiba, Hiroki; Tsumoto, Kouhei

    2015-07-01

    Antibodies (immunoglobulins) bind specific molecules (i.e. antigens) with high affinity and specificity. In order to understand their mechanisms of recognition, interaction analysis based on thermodynamic and kinetic parameters, as well as structure determination is crucial. In this review, we focus on mutational analysis which gives information about the role of each amino acid residue in antibody-antigen interaction. Taking anti-hen egg lysozyme antibodies and several anti-small molecule antibodies, the energetic contribution of hot-spot and non-hot-spot residues is discussed in terms of thermodynamics. Here, thermodynamics of the contribution from aromatic, charged and hydrogen bond-forming amino acids are discussed, and their different characteristics have been elucidated. The information gives fundamental understanding of the antibody-antigen interaction. Furthermore, the consequences of antibody engineering are analysed from thermodynamic viewpoints: humanization to reduce immunogenicity and rational design to improve affinity. Amino acid residues outside hot-spots in the interface play important roles in these cases, and thus thermodynamic and kinetic parameters give much information about the antigen recognition. Thermodynamic analysis of mutant antibodies thus should lead to advanced strategies to design and select antibodies with high affinity. PMID:25956164

  14. Cofactor-containing antibodies: crystal structure of the original yellow antibody.

    PubMed

    Zhu, Xueyong; Wentworth, Paul; Kyle, Robert A; Lerner, Richard A; Wilson, Ian A

    2006-03-01

    Antibodies are generally thought to be a class of proteins that function without the use of cofactors. However, it is not widely appreciated that antibodies are believed to be the major carrier protein in human circulation for the important riboflavin cofactor that is involved in a host of biological phenomena. A further link between riboflavin and antibodies was discovered 30 years ago when a bright-yellow antibody, IgG(GAR), was purified from a patient with multiple myeloma who had turned yellow during the course of her disease. It was subsequently shown that the yellow color of this antibody was due to riboflavin binding. However, it was not known how and where riboflavin was bound to this antibody. We now report the crystal structure of this historically important IgG(GAR) Fab at 3.0-A resolution. The riboflavin is located in the antigen-combining site with its isoalloxazine ring stacked between the parallel aromatic moieties of TyrH33, PheH58, and TyrH100A. Together with additional hydrogen bonds, these interactions reveal the structural basis for high-affinity riboflavin binding. The ligand specificity of IgG(GAR) is compared with another riboflavin-binding antibody, IgG(DOT), which was purified from a second patient with multiple myeloma. The crystal structure of IgG(GAR) provides a starting point for attempts to understand the physiological relevance and chemical functions of cofactor-containing antibodies. PMID:16537445

  15. Panel reactive antibody positivity and associated HLA antibodies in Turkish renal transplant candidates

    Microsoft Academic Search

    Fatma Nurhan Ozdemir; Siren Sezer; Ali Akcay; Zubeyde Arat; Munire Turan; Sale Gulmus; Eyup Kulah; Mehmet Haberal

    2004-01-01

    Pre- and post-renal transplantation panel reactive antibody (PRA) screening is associated with increased incidence of hyperacute or acute graft rejection and graft loss. This study was designed to find any relationship PRA sensitization and associated human leukocyte antigen (HLA)-specific antibodies in Turkish renal transplant candidates. We included 340 patients who were in the renal transplantation waiting list in the study.

  16. Higher cytotoxicity of divalent antibody-toxins than monovalent antibody-toxins

    SciTech Connect

    Won, JaeSeon; Nam, PilWon; Lee, YongChan [College of Life Sciences and Graduate School of Biotechnology, Korea University, 5-ga Anam-dong, Sungbuk-gu, Seoul 136-701 (Korea, Republic of)] [College of Life Sciences and Graduate School of Biotechnology, Korea University, 5-ga Anam-dong, Sungbuk-gu, Seoul 136-701 (Korea, Republic of); Choe, MuHyeon, E-mail: choemh@korea.ac.kr [College of Life Sciences and Graduate School of Biotechnology, Korea University, 5-ga Anam-dong, Sungbuk-gu, Seoul 136-701 (Korea, Republic of)] [College of Life Sciences and Graduate School of Biotechnology, Korea University, 5-ga Anam-dong, Sungbuk-gu, Seoul 136-701 (Korea, Republic of)

    2009-04-24

    Recombinant antibody-toxins are constructed via the fusion of a 'carcinoma-specific' antibody fragment to a toxin. Due to the high affinity and high selectivity of the antibody fragments, antibody-toxins can bind to surface antigens on cancer cells and kill them without harming normal cells [L.H. Pai, J.K. Batra, D.J. FitzGerald, M.C. Willingham, I. Pastan, Anti-tumor activities of immunotoxins made of monoclonal antibody B3 and various forms of Pseudomonas exotoxin, Proc. Natl. Acad. Sci. USA 88 (1991) 3358-3362]. In this study, we constructed the antibody-toxin, Fab-SWn-PE38, with SWn (n = 3, 6, 9) sequences containing n-time repeated (G{sub 4}S) between the Fab fragment and PE38 (38 kDa truncated form of Pseudomonas exotoxin A). The SWn sequence also harbored one cysteine residue that could form a disulfide bridge between two Fab-SWn-PE38 monomers. We assessed the cytotoxicity of the monovalent (Fab-SWn-PE38), and divalent ([Fab-SWn-PE38]{sub 2}) antibody-toxins. The cytotoxicity of the dimer against the CRL1739 cell line was approximately 18.8-fold higher than that of the monomer on the ng/ml scale, which was approximately 37.6-fold higher on the pM scale. These results strongly indicate that divalency provides higher cytotoxicity for an antibody-toxin.

  17. The antibody mining toolbox: an open source tool for the rapid analysis of antibody repertoires.

    PubMed

    D'Angelo, Sara; Glanville, Jacob; Ferrara, Fortunato; Naranjo, Leslie; Gleasner, Cheryl D; Shen, Xiaohong; Bradbury, Andrew R M; Kiss, Csaba

    2014-01-01

    In vitro selection has been an essential tool in the development of recombinant antibodies against various antigen targets. Deep sequencing has recently been gaining ground as an alternative and valuable method to analyze such antibody selections. The analysis provides a novel and extremely detailed view of selected antibody populations, and allows the identification of specific antibodies using only sequencing data, potentially eliminating the need for expensive and laborious low-throughput screening methods such as enzyme-linked immunosorbant assay. The high cost and the need for bioinformatics experts and powerful computer clusters, however, have limited the general use of deep sequencing in antibody selections. Here, we describe the AbMining ToolBox, an open source software package for the straightforward analysis of antibody libraries sequenced by the three main next generation sequencing platforms (454, Ion Torrent, MiSeq). The ToolBox is able to identify heavy chain CDR3s as effectively as more computationally intense software, and can be easily adapted to analyze other portions of antibody variable genes, as well as the selection outputs of libraries based on different scaffolds. The software runs on all common operating systems (Microsoft Windows, Mac OS X, Linux), on standard personal computers, and sequence analysis of 1-2 million reads can be accomplished in 10-15 min, a fraction of the time of competing software. Use of the ToolBox will allow the average researcher to incorporate deep sequence analysis into routine selections from antibody display libraries. PMID:24423623

  18. Baculovirus display of functional antibody Fab fragments.

    PubMed

    Takada, Shinya; Ogawa, Takafumi; Matsui, Kazusa; Suzuki, Tasuku; Katsuda, Tomohisa; Yamaji, Hideki

    2015-08-01

    The generation of a recombinant baculovirus that displays antibody Fab fragments on the surface was investigated. A recombinant baculovirus was engineered so that the heavy chain (Hc; Fd fragment) of a mouse Fab fragment was expressed as a fusion to the N-terminus of baculovirus gp64, while the light chain of the Fab fragment was simultaneously expressed as a secretory protein. Following infection of Sf9 insect cells with the recombinant baculovirus, the culture supernatant was analyzed by enzyme-linked immunosorbent assay using antigen-coated microplates and either an anti-mouse IgG or an anti-gp64 antibody. A relatively strong signal was obtained in each case, showing antigen-binding activity in the culture supernatant. In western blot analysis of the culture supernatant using the anti-gp64 antibody, specific protein bands were detected at an electrophoretic mobility that coincided with the molecular weight of the Hc-gp64 fusion protein as well as that of gp64. Flow cytometry using a fluorescein isothiocyanate-conjugated antibody specific to mouse IgG successfully detected the Fab fragments on the surface of the Sf9 cells. These results suggest that immunologically functional antibody Fab fragments can be displayed on the surface of baculovirus particles, and that a fluorescence-activated cell sorter with a fluorescence-labeled antigen can isolate baculoviruses displaying specific Fab fragments. This successful baculovirus display of antibody Fab fragments may offer a novel approach for the efficient selection of specific antibodies. PMID:25906386

  19. [Characterisation of a monoclonal antibody against Trypanosoma evansi and its application for detecting circulating antibodies].

    PubMed

    Monzón, C M

    2006-12-01

    Monoclonal antibodies were obtained against Trypanosoma evansi. The 2-4F6 IgM monoclonal antibody (Mab) was chosen for the study because of its ability to detect antigens and its specificity (as it did not recognise T. cruzi, T. equiperdum, Babesia equi or B. caballi). The immunoblot test revealed that the 2-4F6 IgM Mab recognises epitopes in two antigenic bands, one measuring 85 kDa and the other 122 kDa. An immunoassay for antigen detection in serum using polyclonal antibodies for capture, the Mab 2-4F6 as primary antibody and an antimouse IgM as secondary antibody gave positive results in 10 of the 11 equidae infected with T. evansi, whereas 20 controls gave negative results. These research results show that the Mab 2-4F6 and the antigen it recognises are useful in identifying equidae infected with T. evansi. PMID:17361770

  20. Antibody engineering by parsimonious mutagenesis.

    PubMed

    Balint, R F; Larrick, J W

    1993-12-27

    The human monoclonal antibody (humAb) problem has largely been solved with the aid of the polymerase chain reaction (PCR) [Larrick et al., Bio/Technology 7 (1989a) 934-938; Larrick et al., Biochem. Biophys. Res. Commun. 160 (1989b) 1250-1256; Chiang et al., BioTechniques 7 (1989) 360-366]. Phage display has now made it possible to recover humAb with primary response level affinities (approx. 10(6) M-1) for virtually any antigen (including self antigens) from comprehensive libraries of B-cell repertoires from non-immunized humans [Marks et al., J. Mol. Biol. 222 (1991) 581-597; Marks et al., Bio/Technology 10 (1992) 779-783; Griffiths et al., EMBO J. 12 (1993) 725-734]. This means that the goal of therapeutic humAb without immunization is within reach. However, in order to achieve the affinities generally required for therapeutic use (> or = 10(9) M-1), reliable methods will be needed to complete the affinity maturation process in vitro. Available X-ray crystallographic data and energy calculations indicate that only a fraction of the substantial contact surface between the Ab and protein antigens contribute significantly to affinity. Thus, the remaining contact surface presents multiple opportunities to develop additional high-affinity contacts, needing only a means to identify them. To this end, we have developed a computer-assisted method for oligodeoxyribonucleotide-directed scanning mutagenesis, called parsimonious mutagenesis (PM), whereby all three complementarity-determining regions (CDR) of a variable region (V-region) gene can be simultaneously and thoroughly searched for improved variants in libraries of manageable size. These libraries are made with low-redundancy 'doping' codons and biased nucleotide (nt) mixtures designed to maximize the abundance of combining sites with predetermined proportions of preselected sets of alternative amino acids (aa). This allows the library to 'probe' the surface of the antigen one or a few aa residues at a time with a wide selection of aa side chains to search out and identify new high-affinity contacts. In addition to affinity maturation in vitro, PM can also be used to remove unwanted cross-reactivities and to 'reshape' rodent mAb for human therapeutic use. PMID:7506686

  1. Smooth muscle antibodies in Mycoplasma pneumoniae infection.

    PubMed Central

    Biberfeld, G; Sterner, G

    1976-01-01

    Paired sera from forty-five cases of Mycoplasma pneumoniae (MP) infection associated with acute lower respiratory tract illness were examined by immunofluorescence for antibodies to smooth muscle. Twenty-five (56%) of these cases had smooth muscle antibodies (SMA) of IgM class. A significant (greater than or equal to 4-fold) increase in titre of these antibodies was demonstrated in fifteen of thirty-five patients with a significant rise in titre of MP antibodies. SMA of IgG class occurred in eleven of forty-five cases (24%), but a 4-fold rise in antibody titre was found only in two cases. Three of forty-five sera (7%) from healthy donors contained SMA of IgM class and eight sera (18%) SMA of IgG class. MP antigen did not absorb SMA. Liver tests were performed in twenty-nine patients. In eighteen patients SGPT values were moderately or slightly elevated. There was no correlation between the occurrence of increased levels of transaminases and the presence of SMA in serum. In a patient with active chronic hepatitis, who had had a high titre of SMA exclusively of IgG class for 2 years, SMA of IgM class appeared transiently in association with an acute respiratory illness due to MP. PMID:1084242

  2. Sensitivity of HIV antibody detection in saliva.

    PubMed

    Stark, K; Warnecke, C; Brinkmann, V; Gelderblom, H R; Bienzle, U; Pauli, G

    1993-07-01

    To assess the sensitivity and specificity of HIV antibody detection in saliva we tested matched serum and saliva samples from HIV-infected and uninfected individuals. Saliva specimens were collected by two different devices of the Salivette system and stored at different temperatures. Samples were tested for HIV antibodies by two commercially available enzyme-linked immunosorbent assays (ELISAs; Wellcome, Biotest). HIV antibodies were detected in 98.5% (Wellcome) and 97.8% (Biotest) of the saliva samples (standard Salivettes) from 135 seropositive individuals. Using the Salivettes flavoured with citric acid the sensitivity was only 22.9%. No reactions in ELISA were found in saliva from HIV-seronegative individuals. Salivary HIV-specific IgA was detected in 90% of seropositive individuals. All positive saliva samples stored at room temperature were still reactive after 20 days; of those stored at 37 degrees C, 23 out of 24 were positive when retested on day 5. Sensitivity of HIV antibody detection in saliva samples dried onto filter paper was 100% when a minimum of 100 microliters of saliva was used. HIV antibody testing in saliva is an efficient tool for large scale epidemiological studies when standard Salivettes are used for sample collection. Saliva samples can be stored in Salivettes or dried onto filter paper for several days at room temperature and under tropical conditions (37 degrees C). PMID:8232068

  3. Pneumococcal vaccine and opsonic pneumococcal antibody.

    PubMed

    Song, Joon Young; Moseley, M Allen; Burton, Robert L; Nahm, Moon H

    2013-06-01

    Streptococcus pneumoniae is a major human pathogen responsible for the majority of bacterial pneumonia cases as well as invasive pneumococcal diseases with high mortality and morbidity. Use of conjugate vaccines targeting the pneumococcal capsule has dramatically reduced the incidence of invasive diseases, and there are active efforts to further improve the conjugate vaccines. However, in children new pneumococcal vaccines can no longer be tested with placebo-based clinical trials because effective vaccines are currently available. Thus, vaccine studies must depend on surrogate markers of vaccine efficacy. Although traditional antibody levels (e.g., ELISA) are useful as a surrogate marker of protection, they have limitations, and a bioassay measuring the capacity of antibodies to opsonize pneumococci has been developed. This opsonophagocytosis assay (OPA) replicates the in vivo mechanism of antibody protection and should therefore better reflect protection by vaccine-induced antibodies. Technical improvements of OPA have made this bioassay rapid, multiplexed, and practical for analyzing small samples including those from children. Strong correlations between ELISA and OPA have been observed in many studies of young children. However, poor correlations have been found in some important clinical situations (such as determination of protection by cross-reactive antibodies) and populations (such as elderly adults and immunodeficient patients). In these settings, OPA has become a useful supplementary measure of pneumococcal vaccine immunogenicity. Current efforts to standardize OPA will further expand its uses. PMID:23657429

  4. Antibody-specific model of amino acid substitution for immunological inferences from alignments of antibody sequences.

    PubMed

    Mirsky, Alexander; Kazandjian, Linda; Anisimova, Maria

    2015-03-01

    Antibodies are glycoproteins produced by the immune system as a dynamically adaptive line of defense against invading pathogens. Very elegant and specific mutational mechanisms allow B lymphocytes to produce a large and diversified repertoire of antibodies, which is modified and enhanced throughout all adulthood. One of these mechanisms is somatic hypermutation, which stochastically mutates nucleotides in the antibody genes, forming new sequences with different properties and, eventually, higher affinity and selectivity to the pathogenic target. As somatic hypermutation involves fast mutation of antibody sequences, this process can be described using a Markov substitution model of molecular evolution. Here, using large sets of antibody sequences from mice and humans, we infer an empirical amino acid substitution model AB, which is specific to antibody sequences. Compared with existing general amino acid models, we show that the AB model provides significantly better description for the somatic evolution of mice and human antibody sequences, as demonstrated on large next generation sequencing (NGS) antibody data. General amino acid models are reflective of conservation at the protein level due to functional constraints, with most frequent amino acids exchanges taking place between residues with the same or similar physicochemical properties. In contrast, within the variable part of antibody sequences we observed an elevated frequency of exchanges between amino acids with distinct physicochemical properties. This is indicative of a sui generis mutational mechanism, specific to antibody somatic hypermutation. We illustrate this property of antibody sequences by a comparative analysis of the network modularity implied by the AB model and general amino acid substitution models. We recommend using the new model for computational studies of antibody sequence maturation, including inference of alignments and phylogenetic trees describing antibody somatic hypermutation in large NGS data sets. The AB model is implemented in the open-source software CodonPhyML (http://sourceforge.net/projects/codonphyml) and can be downloaded and supplied by the user to ProGraphMSA (http://sourceforge.net/projects/prographmsa) or other alignment and phylogeny reconstruction programs that allow for user-defined substitution models. PMID:25534034

  5. Novel human antibody therapeutics: The age of the Umabs

    PubMed Central

    Ruuls, Sigrid R; van Bueren, Jeroen J Lammerts; van de Winkel, Jan G J; Parren, Paul W H I

    2008-01-01

    Monoclonal antibodies represent a major and increasingly important category of biotechnology products for the treatment of human diseases. The state-of-the-art of antibody technology has evolved to the point where therapeutic monoclonal antibodies, that are practically indistinguishable from antibodies induced in humans, are routinely generated. We depict how our science-based approach can be used to further improve the efficacy of antibody therapeutics, illustrated by the development of three monoclonal antibodies for various cancer indications: zanolimumab (directed against CD4), ofatumumab (directed against CD20) and zalutumumab (directed against epidermal growth factor receptor). PMID:18702090

  6. Cross-reactive and pre-existing antibodies to therapeutic antibodies-Effects on treatment and immunogenicity.

    PubMed

    van Schie, Karin A; Wolbink, Gerrit-Jan; Rispens, Theo

    2015-07-01

    The potential for immunogenicity is an ever-present concern during the development of biopharmaceuticals. Therapeutic antibodies occasionally elicit an antibody response in patients, which can result in loss of response or adverse effects. However, antibodies that bind a drug are sometimes found in pre-treatment serum samples, with the amount depending on drug, assay, and patient population. This review summarizes published data on pre-existing antibodies to therapeutic antibodies, including rheumatoid factors, anti-allotype antibodies, anti-hinge antibodies, and anti-glycan antibodies. Unlike anti-idiotype antibodies elicited by the drug, pre-formed antibodies in general appear to have little consequences during treatment. In the few cases where (potential) clinical consequences were encountered, antibodies were characterized and found to bind a distinct, unusual epitope of the therapeutic. Immunogenicity testing strategies should therefore always include a proper level of antibody characterization, especially when pre-formed antibodies are present. This minimizes false-positives, particularly due to rheumatoid factors, and helps to judge the potential threat in case a genuine pre-dose antibody reactivity is identified. PMID:25962087

  7. The expanding role of therapeutic antibodies.

    PubMed

    Salemi, Simonetta; Markovic, Milica; Martini, Gabriella; D'Amelio, Raffaele

    2015-05-01

    Therapeutic antibodies have been used since the end of nineteenth century, but their use is progressively increased and recently, with the availability of monoclonal antibodies, they are successfully employed in a large disease spectrum, which transversally covers different fields of medicine. Hyperimmune polyclonal immune globulin has been used against infectious diseases, in a period in which anti-microbial drugs were not yet available, and it still maintains a relevant place in prophylaxis/therapy. Although immune globulin should be considered life-saving as replacement therapy in humoral immunodeficiencies, its place in the immune-modulating treatment is not usually first-choice, but it should be considered as support to standard approved treatments. Despite therapeutic monoclonal antibodies have been lastly introduced in therapy, their extreme potentiality is reflected by the large number of approved molecules, addressed toward different immunological targets and able to heavily influence the prognosis and quality of life of a wide range of different diseases. PMID:24471447

  8. Removal of Species Constraints in Antibody Detection ?

    PubMed Central

    Basile, Alison Jane; Biggerstaff, Brad J.; Kosoy, Olga L.; Junna, Shilpa R.; Panella, Nicholas A.; Powers, Ann M.; Stark, Lillian M.; Nemeth, Nicole M.

    2010-01-01

    Serum antibodies from myriad species, particularly birds, can provide key information regarding the transmission and the expansion of the territory of emerging pathogens. Expedient antibody analysis is constrained by a lack of species-specific reagents, a deficiency potentially highlighted by the recent swine-origin influenza A virus (H1N1) outbreak. Available methodologies present difficulties that discourage thorough serologic monitoring of potential disease vectors or hosts. Rapid high-throughput procedures that combined serum amine labeling via biotinylation, contaminant removal, and microsphere-based immunoassays for antibodies to three arboviruses were developed. Agent-specific adaptations of this simple format should facilitate expanded surveillance and diagnostic capabilities regarding pathogens of human and veterinary importance. PMID:19923570

  9. Unnatural amino acids in novel antibody conjugates.

    PubMed

    Hallam, Trevor J; Smider, Vaughn V

    2014-07-01

    Antibody-drug conjugates are an important and emerging drug class for the treatment of cancer. Recent evidence strongly suggests that site-specific drug conjugation results in a homogenous population of molecules with more favorable activity and pharmacokinetic properties than randomly conjugated antibodies. Unnatural amino acids (uAAs) can be incorporated in recombinant proteins to enable unique orthogonal chemistries in comparison to the side chains of the natural 20 amino acids. Thus, uAAs present a novel platform for which to create next-generation antibody-drug conjugates. Furthermore, site-specific conjugation through uAAs can also enpower unique small molecule, bispecific, multispecific and other conjugates that could be important constructs for therapeutics, diagnostics and research reagents. Here, we review the progress in uAA incorporation and conjugate construction through both cell-based and -free approaches. PMID:25163001

  10. Monoclonal antibodies specific for sickle cell hemoglobin

    SciTech Connect

    Jensen, R.H.; Vanderlaan, M.; Grabske, R.J.; Branscomb, E.W.; Bigbee, W.L.; Stanker, L.H.

    1985-01-01

    Two mouse hybridoma cell lines were isolated which produce monoclonal antibodies that bind hemoglobin S. The mice were immunized with peptide-protein conjugates to stimulate a response to the amino terminal peptide of the beta chain of hemoglobin S, where the single amino acid difference between A and S occurs. Immunocharacterization of the antibodies shows that they bind specifically to the immunogen peptide and to hemoglobin S. The specificity for S is high enough that one AS cell in a mixture with a million AA cells is labeled by antibody, and such cells can be analyzed by flow cytometry. Immunoblotting of electrophoretic gels allows definitive identification of hemoglobin S as compared with other hemoglobins with similar electrophoretic mobility. 12 references, 4 figures.

  11. Is antenatal antibody screening worthwhile in Chinese?

    PubMed

    Wong, K F; Tse, K T; Lee, A W; Mak, C S; So, C C

    1997-06-01

    A total of 1997 pregnant women were screened during their first antenatal visit for irregular antibodies for the prevention of haemolytic disease of the newborn. Patient sera were tested against a panel of group O screen cells including one with the expression of Miltenberger determinants GP.Mur. 17 women (0.85%) had irregular antibodies of which four were of potential clinical significance, including one with anti-D, two with anti-E and one with anti-D, anti-E and anti-G. Although antenatal antibody screening is mandatory in Western populations, our results suggest that this may not be necessary in the Chinese population except for those who are Rh D-negative or who have a history of haemolytic disease of the newborn. PMID:9217197

  12. An immunosuppressive antibody-drug conjugate.

    PubMed

    Wang, Rongsheng E; Liu, Tao; Wang, Ying; Cao, Yu; Du, Jintang; Luo, Xiaozhou; Deshmukh, Vishal; Kim, Chan Hyuk; Lawson, Brian R; Tremblay, Matthew S; Young, Travis S; Kazane, Stephanie A; Wang, Feng; Schultz, Peter G

    2015-03-11

    We have developed a novel antibody-drug conjugate (ADC) that can selectively deliver the Lck inhibitor dasatinib to human T lymphocytes. This ADC is based on a humanized antibody that selectively binds with high affinity to CXCR4, an antigen that is selectively expressed on hematopoietic cells. The resulting dasatinib-antibody conjugate suppresses T-cell-receptor (TCR)-mediated T-cell activation and cytokine expression with low nM EC50 and has minimal effects on cell viability. This ADC may lead to a new class of selective immunosuppressive drugs with improved safety and extend the ADC strategy to the targeted delivery of kinase inhibitors for indications beyond oncology. PMID:25699419

  13. [Demonstration of trichinella antibodies (author's transl)].

    PubMed

    Saathoff, M; Kasper, M; Demmer, H

    1978-10-13

    Serum samples of humans and animals with Trichinella infection of different duration and severity were analysed for Trichinella antibodies by four different serological methods. The indirect immunofluorescence test and the indirect haemagglutination test proved to be the most sensitive ones. Complement-fixation reaction gave the poorest results. In fresh Trichinella infections the heterologous antibody against cercaria of S. mansoni was more strongly developed than the homologous antibody which is effective in the microprecision test on Trichinella larvae. Although the animals deliberately infected also had, at the beginning of infection, a marked reaction to cercaria, this test became negative earlier than the microprecipitation test. Differentiation of Trichinella from Schistosoma infections is serologically possible without difficulty by means of adult schistosomes. The cross-reaction observed between T. spiralis and various Filaria types can be differentiated with adult O. volvulus as antigen. PMID:359293

  14. Current status of antibody therapy in ALL.

    PubMed

    Ai, Jing; Advani, Anjali

    2015-02-01

    Despite the significant advances in modern chemotherapy, it remains challenging to treat adult patients with acute lymphoblastic leukaemia (ALL). The relapse rate remains high, and the outcome at the time of relapse is dismal. Antibody-based therapies have demonstrated promising results in this patient group. Variable mechanisms have been applied to target surface antigens (CD20 [also termed MS4A1], CD22, CD52 and CD19) that are commonly expressed on malignant leukaemia cells. In this review, we will focus on the clinical application of such therapies in adult ALL, including the naked antibodies: Rituximab, Ofatumumab, Epratuzumab and Alemtuzumab; the immunotoxins: BL22 and Combotox; the immunoconjugates: inotuzumab and SAR 3419; as well as the Bi-specific T cell engaging (BiTE)-specific antibody, Blinatumomab. PMID:25382151

  15. Engineered antibodies for molecular imaging of cancer

    PubMed Central

    Wu, Anna M.

    2013-01-01

    Antibody technology has transformed drug development, providing robust approaches to producing highly targeted and active therapeutics that can routinely be advanced through clinical evaluation and registration. In parallel, there is an emerging need to access similarly targeted agents for diagnostic purposes, including non-invasive imaging in preclinical models and patients. Antibody engineering enables modification of key properties (immunogenicity, valency, biological inertness, pharmacokinetics, clearance route, site-specific conjugation) in order to produce targeting agents optimized for molecular imaging. Expanded availability of positron-emitting radionuclides has led to a resurgence of interest and applications of immunoPET (immuno-positron emission tomography). Molecular imaging using engineered antibodies and fragments provides a general approach for assessing cell surface phenotype in vivo and stands to play an increasingly important role in cancer diagnosis, treatment selection, and monitoring of molecularly targeted therapeutics. PMID:24091005

  16. THE MODIFICATION OF ANTIBODIES BY FORMALDEHYDE

    PubMed Central

    Mudd, Stuart; Joffe, Eleanore W.

    1933-01-01

    Certain strains of bacteria which have only minimal zeta potentials over a wide range of pH, and upon which surface deposits can be formed, afford a favorable means of studying certain chemical and physical properties of the surface deposits. Films of specific antibody-globulin upon these bacteria possess basic groups which can combine with formaldehyde. Combination of these groups with HCHO under the conditions of the present experiments shifts the isoelectric point of the sensitizing film toward the acid side by about 0.6 to 0.8 pH unit, and reduces the agglutinating tendency of the sensitizing film. Antibodies may be formalinized before combination with antigen without marked change in their specific combining affinities. The properties of the sensitizing films are similar whether formol treatment occurs before or after the antigen-antibody combination. The nature of the basic groups has been discussed. PMID:19872752

  17. Removal of species constraints in antibody detection.

    PubMed

    Basile, Alison Jane; Biggerstaff, Brad J; Kosoy, Olga L; Junna, Shilpa R; Panella, Nicholas A; Powers, Ann M; Stark, Lillian M; Nemeth, Nicole M

    2010-01-01

    Serum antibodies from myriad species, particularly birds, can provide key information regarding the transmission and the expansion of the territory of emerging pathogens. Expedient antibody analysis is constrained by a lack of species-specific reagents, a deficiency potentially highlighted by the recent swine-origin influenza A virus (H1N1) outbreak. Available methodologies present difficulties that discourage thorough serologic monitoring of potential disease vectors or hosts. Rapid high-throughput procedures that combined serum amine labeling via biotinylation, contaminant removal, and microsphere-based immunoassays for antibodies to three arboviruses were developed. Agent-specific adaptations of this simple format should facilitate expanded surveillance and diagnostic capabilities regarding pathogens of human and veterinary importance. PMID:19923570

  18. Multiplex serology of paraneoplastic antineuronal antibodies.

    PubMed

    Maat, Peter; Brouwer, Eric; Hulsenboom, Esther; VanDuijn, Martijn; Schreurs, Marco W J; Hooijkaas, Herbert; Smitt, Peter A E Sillevis

    2013-05-31

    Paraneoplastic neurological syndromes (PNS) are devastating neurological disorders secondary to cancer, associated with onconeural autoantibodies. Such antibodies are directed against neuronal antigens aberrantly expressed by the tumor. The detection of onconeural antibodies in a patient is extremely important in diagnosing a neurological syndrome as paraneoplastic (70% is not yet known to have cancer) and in directing the search for the underlying neoplasm. At present six onconeural antibodies are considered 'well characterized' and recognize the antigens HuD, CDR62 (Yo), amphiphysin, CRMP-5 (CV2), NOVA-1 (Ri), and Ma2. The gold standard of detection is the characteristic immunohistochemical staining pattern on brain tissue sections combined with confirmation by immunoblotting using recombinant purified proteins. Since all six onconeural antibodies are usually analyzed simultaneously and objective cut-off values for these analyses are warranted, we developed a multiplex assay based on Luminex technology. Reaction of serial dilutions of six onconeural standard sera with microsphere-bound antigens showed lower limits of detection than with Western blotting. Using the six standard sera at a dilution of 1:200, the average within-run coefficient of variation (CV) was 4% (range 1.9-7.3%). The average between-run within-day CV was 5.1% (range 2.9-6.7%) while the average between-day CV was 8.1% (range 2.8-11.6%). The shelf-life of the antigen coupled microspheres was at least two months. The sensitivity of the multiplex assay ranged from 83% (Ri) to 100% (Yo, amphiphysin, CV2) and the specificity from 96% (CV2) to 100% (Ri). In conclusion, Luminex-based multiplex serology is highly reproducible with high sensitivity and specificity for the detection of onconeural antibodies. Conventional immunoblotting for diagnosis of onconeural antibodies in the setting of a routine laboratory may be replaced by this novel, robust technology. PMID:23500780

  19. Proteomic Identification of Monoclonal Antibodies from Serum

    PubMed Central

    2015-01-01

    Characterizing the in vivo dynamics of the polyclonal antibody repertoire in serum, such as that which might arise in response to stimulation with an antigen, is difficult due to the presence of many highly similar immunoglobulin proteins, each specified by distinct B lymphocytes. These challenges have precluded the use of conventional mass spectrometry for antibody identification based on peptide mass spectral matches to a genomic reference database. Recently, progress has been made using bottom-up analysis of serum antibodies by nanoflow liquid chromatography/high-resolution tandem mass spectrometry combined with a sample-specific antibody sequence database generated by high-throughput sequencing of individual B cell immunoglobulin variable domains (V genes). Here, we describe how intrinsic features of antibody primary structure, most notably the interspersed segments of variable and conserved amino acid sequences, generate recurring patterns in the corresponding peptide mass spectra of V gene peptides, greatly complicating the assignment of correct sequences to mass spectral data. We show that the standard method of decoy-based error modeling fails to account for the error introduced by these highly similar sequences, leading to a significant underestimation of the false discovery rate. Because of these effects, antibody-derived peptide mass spectra require increased stringency in their interpretation. The use of filters based on the mean precursor ion mass accuracy of peptide-spectrum matches is shown to be particularly effective in distinguishing between “true” and “false” identifications. These findings highlight important caveats associated with the use of standard database search and error-modeling methods with nonstandard data sets and custom sequence databases. PMID:24684310

  20. Quantitative cumulative biodistribution of antibodies in mice

    PubMed Central

    Yip, Victor; Palma, Enzo; Tesar, Devin B; Mundo, Eduardo E; Bumbaca, Daniela; Torres, Elizabeth K; Reyes, Noe A; Shen, Ben Q; Fielder, Paul J; Prabhu, Saileta; Khawli, Leslie A; Boswell, C Andrew

    2014-01-01

    The neonatal Fc receptor (FcRn) plays an important and well-known role in antibody recycling in endothelial and hematopoietic cells and thus it influences the systemic pharmacokinetics (PK) of immunoglobulin G (IgG). However, considerably less is known about FcRn’s role in the metabolism of IgG within individual tissues after intravenous administration. To elucidate the organ distribution and gain insight into the metabolism of humanized IgG1 antibodies with different binding affinities FcRn, comparative biodistribution studies in normal CD-1 mice were conducted. Here, we generated variants of herpes simplex virus glycoprotein D-specific antibody (humanized anti-gD) with increased and decreased FcRn binding affinity by genetic engineering without affecting antigen specificity. These antibodies were expressed in Chinese hamster ovary cell lines, purified and paired radiolabeled with iodine-125 and indium-111. Equal amounts of I-125-labeled and In-111-labeled antibodies were mixed and intravenously administered into mice at 5 mg/kg. This approach allowed us to measure both the real-time IgG uptake (I-125) and cumulative uptake of IgG and catabolites (In-111) in individual tissues up to 1 week post-injection. The PK and distribution of the wild-type IgG and the variant with enhanced binding for FcRn were largely similar to each other, but vastly different for the rapidly cleared low-FcRn-binding variant. Uptake in individual tissues varied across time, FcRn binding affinity, and radiolabeling method. The liver and spleen emerged as the most concentrated sites of IgG catabolism in the absence of FcRn protection. These data provide an increased understanding of FcRn’s role in antibody PK and catabolism at the tissue level. PMID:24572100

  1. Antibodies to citrullinated peptides in tuberculosis.

    PubMed

    Lima, I; Oliveira, R C; Atta, A; Marchi, S; Barbosa, L; Reis, E; Reis, M G; Santiago, M B

    2013-05-01

    Rheumatoid arthritis (RA) is an autoimmune disease characterized by symmetric polyarthritis, rheumatoid factor (RF) positivity, and bone erosions. Recently, research has been conducted on anti-citrullinated peptide antibodies (ACPAs) to which there are greater sensitivity and specificity than RF. However, these antibodies have also been described in infectious diseases, particularly tuberculosis (TB), placing the high specificity of the test in doubt. The aim of this research was to study the prevalence of ACPAs in TB, RA, and healthy controls. Patients with bacteriologically confirmed pulmonary tuberculosis, RA (ACR criteria), in addition to healthy controls were included. ACPAs were researched by: anti-cyclic citrullinated peptide (CCP), anti-modified citrullinated vimentin (MCV), and RF by ELISA. The study was conducted in 50 TB patients, 50 with RA, and 20 controls. Anti-CCP antibodies were found in 39 (78 %) of the RA patients (median titer, 128 U), whereas anti-MCV antibodies were found in 25 (50 %). Of the patients with TB, two (4 %) had positivity for anti-CCP and anti-MCV and no patient in the control group tested positive for these antibodies. Sensitivity of anti-CCP for RA was 78 % (confidence interval (CI), 63 to 88 %) and specificity was 97 % (CI, 89 to 99 %) while the sensitivity of anti-MCV was 50 % (CI, 35-64 %) and specificity was 97 % (CI, 89 to 99 %). RF was positive in 40 samples (80 %) of RA, in 30 (60 %) of TB, and in 1 (5 %) of the controls. Our findings showed high sensitivity of anti-CCP and high specificity of both anti-CCP and anti-MCV antibodies for RA, even in a population with high incidence of tuberculosis. The higher frequency of positivity of ACPA in TB observed in previous studies may be attributed to methodological factors. PMID:23344687

  2. GAMMA GLOBULIN AND ANTIBODY FORMATION IN VITRO

    PubMed Central

    Thorbecke, G. J.; Asofsky, R. M.; Hochwald, G. M.; Siskind, G. W.

    1962-01-01

    Antibody formation in vitro by red and white pulp of the spleen and by bone marrow tissue was studied at various days after an intravenous booster injection of soluble antigens such as ovalbumin and bovine gamma globulin (BGG). When the booster injection of antigen was given early (10 days) after an intravenous primary injection, high antibody formation could be demonstrated in the spleen primarily 2 to 3 days after the injection, but much less afterwards. When the booster injection was given later (1 month) after the primary, the antibody production by the spleen lasted longer and higher serum titers were obtained. The bone marrow formed antibody in both cases but, particularly with the short interval between injections, its response was delayed as compared to the spleen. It was also shown that during antibody formation the production of gamma globulin in vitro was enhanced. Histologically the antibody production was always correlated to immature plasma cell proliferation, located at the border of red and white pulp and in the red pulp of the spleen. When endotoxin had been injected at the time of a primary BGG injection, and a second antigen injection was given 5 to 10 days later, a booster response could be elicited which was sometimes limited to the white pulp on day 1, and on day 2 was divided between "red" and "white" pulp. The response induced at day 10, at the peak of secondary nodule proliferation, lasted very long and was accompanied by an enormous plasma cellular proliferation in and around the periarteriolar lymphoid areas of the spleen. The possible importance of the secondary nodules of the white pulp in the preparation for a secondary response is discussed. PMID:13920994

  3. Acute antibody-mediated renal allograft rejection associated with HLA-Cw17 antibody

    PubMed Central

    Kuppachi, Sarat

    2012-01-01

    Detection of donor-specific human leukocyte antigen (HLA) antibodies is an important part of diagnosis of antibody-mediated rejection (AMR) in the renal transplant population. Donor-specific antibodies (DSA) against HLA-C, a Class 1 major histocompatibility gene product, are not considered to be of major importance in renal transplant rejection. Typing for HLA-C is not a routine part of pre- and post-transplant evaluation. In roughly 10% of biopsy-proven C4d-positive rejections, DSA are not detected by standard testing protocols. In some of these cases, minor HLA and non-HLA antibodies have been implicated. The role of HLA-C antibodies in this patient group is not clear. We present a patient with acute renal graft dysfunction 21 months post-transplant. The allograft biopsy showed features of AMR with diffuse margination of inflammatory cells and diffuse C4d staining in peritubular capillaries. HLA-Cw17 antibody was detected by single-bead antigen Luminex assay, which was further confirmed by a mock flow crossmatch. This case highlights the importance of checking anti-HLA-Cw antibodies in patients with AMR and no detectable DSA using standard methods.

  4. Gangliosides, Ab1 and Ab2 antibodies

    Microsoft Academic Search

    Alejandro López-Requena; Cristina Mateo De Acosta; Ernesto Moreno; Majela González; Yaquelin Puchades; Ariel Talavera; Nelson Santiago Vispo; Ana María Vázquez; Rolando Pérez

    2007-01-01

    This report is focused on the molecular basis for the interaction of a monoclonal antibody (mAb) and its anti-idiotypic mAb. P3 mAb (Ab1) recognizes N-glycolyl-gangliosides, and 1E10 mAb is one of its anti-idiotypic mAbs (Ab2). Chimeric versions of both antibodies retained their specificity. Charged residues in their H-CDRs, particularly H-CDR3, were considered to play a major role in their binding

  5. Effects of interferon on antibody formation

    NASA Technical Reports Server (NTRS)

    Sonnenfeld, G.

    1984-01-01

    Studies of the effects of interferon on primary and secondary antibody responses and of the relationship of interferon to other cytokines, or cell products, are presented. Dosage- and timing-dependent immunoenhancing and immunosuppressive activities of interferon are documented for mouse spleen cell cultures and for mice infected with murine hepatitis virus (MHV-3). A possibility that altered interferon production might lead to immunopathological disorders, such as lupus erythematosus, AIDS, arthritis, etc., is discussed. Latest technological developments are presented that indicate that interferon does apparently play a major role in the regulation of antibody responses.

  6. Antibodies for Treatment of Clostridium difficile Infection

    PubMed Central

    Wilcox, Mark H.

    2014-01-01

    Antibodies for the treatment of Clostridium difficile infection (CDI) have been demonstrated to be effective in the research and clinical environments. Early uncertainties about molecular and treatment modalities now appear to have converged upon the systemic dosing of mixtures of human IgG1. Although multiple examples of high-potency monoclonal antibodies (MAbs) exist, significant difficulties were initially encountered in their discovery. This minireview describes historical and contemporary MAbs and highlights differences between the most potent MAbs, which may offer insight into the pathogenesis and treatment of CDI. PMID:24789799

  7. Efficient generation of monoclonal antibodies from single rhesus macaque antibody secreting cells.

    PubMed

    Meng, Weixu; Li, Leike; Xiong, Wei; Fan, Xuejun; Deng, Hui; Bett, Andrew J; Chen, Zhifeng; Tang, Aimin; Cox, Kara S; Joyce, Joseph G; Freed, Daniel C; Thoryk, Elizabeth; Fu, Tong-Ming; Casimiro, Danilo R; Zhang, Ningyan; A Vora, Kalpit; An, Zhiqiang

    2015-07-01

    Nonhuman primates (NHPs) are used as a preclinical model for vaccine development, and the antibody profiles to experimental vaccines in NHPs can provide critical information for both vaccine design and translation to clinical efficacy. However, an efficient protocol for generating monoclonal antibodies from single antibody secreting cells of NHPs is currently lacking. In this study we established a robust protocol for cloning immunoglobulin (IG) variable domain genes from single rhesus macaque (Macaca mulatta) antibody secreting cells. A sorting strategy was developed using a panel of molecular markers (CD3, CD19, CD20, surface IgG, intracellular IgG, CD27, Ki67 and CD38) to identify the kinetics of B cell response after vaccination. Specific primers for the rhesus macaque IG genes were designed and validated using cDNA isolated from macaque peripheral blood mononuclear cells. Cloning efficiency was averaged at 90% for variable heavy (VH) and light (VL) domains, and 78.5% of the clones (n = 335) were matched VH and VL pairs. Sequence analysis revealed that diverse IGHV subgroups (for VH) and IGKV and IGLV subgroups (for VL) were represented in the cloned antibodies. The protocol was tested in a study using an experimental dengue vaccine candidate. About 26.6% of the monoclonal antibodies cloned from the vaccinated rhesus macaques react with the dengue vaccine antigens. These results validate the protocol for cloning monoclonal antibodies in response to vaccination from single macaque antibody secreting cells, which have general applicability for determining monoclonal antibody profiles in response to other immunogens or vaccine studies of interest in NHPs. PMID:25996084

  8. Long-term vaginal antibody delivery: delivery systems and biodistribution.

    PubMed

    Saltzman, W M; Sherwood, J K; Adams, D R; Castle, P; Haller, P

    2000-02-01

    Topical delivery systems can provide prolonged delivery of antibodies to the vaginal mucosal surface for long-term protection against infectious diseases. We examined the biodistribution of antibodies during 30 days of vaginal antibody delivery in mice. Different antibody preparations (including monoclonal IgG and IgM, as well as several different (125)I-labeled IgGs) were administered by polymer vaginal rings, which were designed to provide continuous antibody delivery. Antibody concentrations remained high in the vaginal secretions for up to 30 days after disk insertion; radiolabeled antibody was also found, at approximately 100 times lower concentration, in the blood and other tissues. The measured concentrations agreed reasonably well with a simple pharmacokinetic model, which was used to calculate mucosal and systemic concentrations as a function of antibody delivery and elimination rates. Results from the model were consistent with previously reported antibody pharmacokinetic measurements: the half-life for antibody elimination for the vagina was approximately 3 h; the half-life for IgG(1) clearance from the blood was >1 day; and the overall permeability constant for vaginal uptake of IgG was approximately 0.01 to 0.03 h(-1). These results provide important information for the design of controlled antibody delivery devices for vaginal use, and suggest that high-dose, long-term vaginal administration of antibodies may be a reasonable approach for achieving sustained mucosal and systemic antibody levels. PMID:10620255

  9. Enhancing antibody: a novel component of the immune response.

    PubMed Central

    Nemazee, D A; Sato, V L

    1982-01-01

    Current descriptions of the immune response identify two classes of antigenic stimuli that result in the production of specific antibody: (i) exogenous antigens and (ii) endogenous variable-region determinants of the immune system. We expand this scheme to include a third class of antigenic stimulus--new determinants created by the binding of antibody to antigen. This paper describes a set of monoclonal antibodies which arose after repeated immunization with antigen alone but which bound antibody--antigen complexes. These antibodies recognize determinants on the antibody portion of the complexes that were expressed as a consequence of antigen binding. Antibodies of this general type, "enhancing antibodies," which can strengthen antibody--antigen and idiotypic-anti-idiotypic antibody interactions, may play important regulatory and effector roles in the immune response. We suggest a model that predicts the occurrence and specificity of different classes of such antibodies and provides a conceptual framework that gives a straightforward explanation of the appearance in the immune response of rheumatoid antibodies and of antibodies that bind cooperatively to antigen. Images PMID:6179088

  10. Anti Rh Hemolytic Disease due to Anti C Antibody: Is Testing for Anti D Antibodies Enough?

    PubMed

    Negi, Gita; Singh, Gaur Dushyant

    2012-06-01

    Rh blood group system is a complex blood group system. Rh antibodies are produced in Rh negative individuals following exposure to foreign RBCs after transfusion or pregnancy. Anti C is a rare cause of hemolytic disease of newborn and is very scarcely reported in the literature. The aim of the present case report of Hemolytic disease caused by Anti C antibody is to bring out the fact that antibodies other than anti D should be considered in cases that give a suggestive history but no evidence of Anti D. PMID:23730022

  11. Neutralizing antibodies to HIV-1 induced by immunization

    PubMed Central

    McCoy, Laura E.

    2013-01-01

    Most neutralizing antibodies act at the earliest steps of viral infection and block interaction of the virus with cellular receptors to prevent entry into host cells. The inability to induce neutralizing antibodies to HIV has been a major obstacle to HIV vaccine research since the early days of the epidemic. However, in the past three years, the definition of a neutralizing antibody against HIV has been revolutionized by the isolation of extremely broad and potent neutralizing antibodies from HIV-infected individuals. Considerable hurdles remain for inducing neutralizing antibodies to a protective level after immunization. Meanwhile, novel technologies to bypass the induction of antibodies are being explored to provide prophylactic antibody-based interventions. This review addresses the challenge of inducing HIV neutralizing antibodies upon immunization and considers notable recent advances in the field. A greater understanding of the successes and failures for inducing a neutralizing response upon immunization is required to accelerate the development of an effective HIV vaccine. PMID:23401570

  12. NCI Requests Targets for Monoclonal Antibody Production and Characterization

    Cancer.gov

    In an effort to provide well-characterized monoclonal antibodies to the scientific community, NCI's Antibody Characterization Program requests cancer-related protein targets for affinity production and distribution.

  13. NCI Requests Cancer Targets for Monoclonal Antibody Production and Characterization

    Cancer.gov

    In an effort to provide well-characterized monoclonal antibodies to the scientific community, NCI's Antibody Characterization Program requests cancer-related protein targets for affinity production and distribution.

  14. Antibody-based therapies for emerging infectious diseases.

    PubMed Central

    Casadevall, A.

    1996-01-01

    In the 19th century, it was discovered that immune sera were useful in treating infectious diseases. Serum therapy was largely abandoned in the 1940s because of the toxicity associated with the administration of heterologous sera and the introduction of effective antimicrobial chemotherapy. Recent advances in the technology of monoclonal antibody production provide the means to generate human antibody reagents and reintroduce antibody therapies, while avoiding the toxicities associated with serum therapy. Because of the versatility of antibodies, antibody-based therapies could, in theory, be developed against any existing pathogen. The advantages of antibody-based therapies include versatility, low toxicity, pathogen specificity, enhancement of immune function, and favorable pharmacokinetics; the disadvantages include high cost, limited usefulness against mixed infections, and the need for early and precise microbiologic diagnosis. The potential of antibodies as antiinfective agents has not been fully tapped. Antibody-based therapies constitute a potentially useful option against newly emergent pathogens. PMID:8903230

  15. Elicitation of structure-specific antibodies by epitope scaffolds

    E-print Network

    Baker, David

    demonstrate the elicitation of structure- specific antibodies against the HIV-1 gp41 epitope of the broadly. The ability of structural biology to provide atomic-level definition of antibody­ antigen interactions

  16. Hypoglycaemic syndromes and antibodies to pancreatic islet cells.

    PubMed Central

    Di Mario, U; Tamburrano, G; Iavicoli, M; Irvine, W J

    1977-01-01

    The sera of ten patients with unexplained hypoglycaemia were examined for antibodies to pancreatic islets. Antibodies to pancreatic A, B and D cells (ICAb) were detected in one patient with an associated gastrointestinal tumour. PMID:338222

  17. Screening individual hybridomas by microengraving to discover monoclonal antibodies

    E-print Network

    Ogunniyi, Adebola Oluwakayode

    The demand for monoclonal antibodies (mAbs) in biomedical research is significant, but the current methodologies used to discover them are both lengthy and costly. Consequently, the diversity of antibodies available for ...

  18. Quantitative analysis of perivascular antibody distribution in solid tumors

    E-print Network

    Rhoden, John J. (John Joseph)

    2013-01-01

    Monoclonal antibodies and proteins derived from them are an emerging class of anticancer therapeutics that have shown efficacy in a range of blood and solid tumors. Antibodies targeting solid tumors face considerable ...

  19. Engineering aglycosylated antibody variants with immune effector functions

    E-print Network

    Sazinsky, Stephen L. (Stephen Lael)

    2009-01-01

    Monoclonal antibodies have emerged as a promising class of therapeutics for the treatment of human disease, and in particular human cancer. While multiple mechanisms contribute to antibody efficacy, the engagement and ...

  20. CiteAb: a searchable antibody database that ranks antibodies by the number of times they have been cited

    PubMed Central

    2014-01-01

    Background Research antibodies are used by thousands of scientists working in diverse disciplines, but it is common to hear concerns about antibody quality. This means that researchers need to carefully choose the antibodies they use to avoid wasting time and money. A well accepted way of selecting a research antibody is to identify one which has been used previously, where the associated data has been peer-reviewed and the results published. Description CiteAb is a searchable database which ranks antibodies by the number of times they have been cited. This allows researchers to easily find antibodies that have been used in peer-reviewed publications and the accompanying citations are listed, so users can check the data contained within the publications. This makes CiteAb a useful resource for identifying antibodies for experiments and also for finding information to demonstrate antibody validation. The database currently contains 1,400,000 antibodies which are from 90 suppliers, including 87 commercial companies and 3 academic resources. Associated with these antibodies are 140,000 publications which provide 306,000 antibody citations. In addition to searching, users can also browse through the antibodies and add their own publications to the CiteAb database. Conclusions CiteAb provides a new way for researchers to find research antibodies that have been used successfully in peer-reviewed publications. It aims to assist these researchers and will hopefully help promote progress in many areas of life science research. PMID:24528853

  1. Boronated monoclonal antibody conjugates for neutron capture therapy

    SciTech Connect

    Borg, D.C.; Elmore, J.J. Jr.; Ferrone, S.

    1986-01-01

    This paper describes the effectiveness of /sup 10/B-labeled monoclonal antibodies against Colo-38 human melanoma in vitro. The authors obtained high boron to antibody ratios while maintaining antibody activity by using dextran intermediate carriers to link /sup 10/B to the antibody. They developed a double cell quasi-competitive binding bioassay to minimize the effects of nonspecific binding of boronated complexes to cells. 1 fig., 2 tabs.

  2. Doxorubicin Conjugates of Monoclonal Antibodies to Hepatoma-Associated Antigens

    Microsoft Academic Search

    Daniel Shouval; Ruth Adler; Jack R. Wands; Esther Hurwitz; Kurt J. Isselbacher; Michael Sela

    1988-01-01

    A panel of six murine monoclonal antibodies against hepatocellular carcinoma-associated antigens, reactive with PLC\\/PRF\\/5 human hepatoma cells, was conjugated to Adriamycin (doxorubicin) via a dextran bridge. This library of antibodies includes three monoclonal antibodies against hepatitis B virus surface antigen, one anti-alpha -fetoprotein, and two other IgG2a antibodies against PLC\\/PRF\\/5 hepatoma-associated antigens. The use of dextran for conjugation of Adriamycin

  3. Antigen-Antibody Testing: A Visual Simulation or Virtual Reality

    NSDL National Science Digital Library

    Daniel L. Schadler (Oglethorpe University; )

    2003-02-24

    This exercise demonstrates the biological phenomenon of the formation of a precipitate when an antigen reacts with an antibody. The exercise can be used to illustrate the specificity of antigen-antibody reactions, showing that a precipitation reaction only occurs when an antibody reacts with the antigen that was used to induce the formation of the antibody. The exercise is also a general demonstration of diffusion.

  4. Immunostimulatory monoclonal antibodies for cancer therapy

    Microsoft Academic Search

    Sandra Hervas-Stubbs; Martin Glennie; Drew M. Pardoll; Ignacio Melero; Lieping Chen

    2007-01-01

    Increasing immune responses with immunostimulatory monoclonal antibodies (mAbs) directed to immune-receptor molecules is a new and exciting strategy in cancer therapy. This expanding class of agents functions on crucial receptors, either antagonizing those that suppress immune responses or activating others that amplify immune responses. Complications such as autoimmunity and systemic inflammation are problematic side effects associated with these agents. However,

  5. World Antibody Drug Conjugate Summit Europe

    PubMed Central

    2011-01-01

    The World Antibody Drug Conjugate Summit Europe, organized by Biorbis/Hanson Wade was held in Frankfurt, Germany February 21–23, 2011. Antibody drug conjugates (ADCs), also called immunoconjugates, are becoming an increasingly important class of therapeutics as demonstrated by the attendance of nearly 100 delegates at this highly focused meeting. Updates on three ADCs that are in late-stage clinical development, trastuzumab emtansine (T-DM1), brentuximab vedotin (SGN-35) and inotuzumab ozogamicin (CMC-544), were presented by speakers from ImmunoGen, Genentech, Roche, Seattle Genetics and Pfizer. These ADCs have shown encouraging therapeutic effects against solid tumors (T-DM1) and hematological malignancies (SGN-35, CMC-544). The key feature of the new generation of ADCs is the effective combination of the cytotoxicity of natural or synthetic highly potent antineoplastic agents, tumor selective monoclonal antibodies and blood-stable optimized linkers. Early clinical data for ADCs were showcased by Progenics Pharmaceuticals (PSMA ADC), Celldex (CDX-011) and Biotest (BT-062). Takeda, MedImmune and sanofi-aventis outlined their strategies for process development and analytical characterization. In addition, presentations on duocarmycin based-ADCs, ? emitting immunoconjugates and antibody-conjugated nanoparticles were given by representatives from Syntarga, Algeta and the University of Stuttgart, respectively. PMID:21691144

  6. Pregnancy outcome in women with antiphospholipid antibodies

    Microsoft Academic Search

    A. K. M. Al-Momen; S. A. Moghraby; M. O. Gad El-Rab; A. M. A. Gader; S. R. Al-Balla; A. A. Al-Meshari; L. Al-Nuaim

    1993-01-01

    The association of antiphospholipid antibodies (APA) or lupus anticoagulant (LA) and recurrent fetal loss (RFL) is well established; however, the spectrum of pregnancy outcome in relation to various therapeutic approaches versus placebo is unknown. We studied 49 women with RFL, 14 with immune thrombocytopenia (ITP) 13 of whom without a history of RFL, and 32 controls (all in the first

  7. Antibody-catalyzed anaerobic destruction of methamphetamine.

    PubMed

    Xu, Yang; Hixon, Mark S; Yamamoto, Noboru; McAllister, Laura A; Wentworth, Anita D; Wentworth, Paul; Janda, Kim D

    2007-03-01

    Methamphetamine [(+)-2] abuse has emerged as a fast-rising global epidemic, with immunopharmacotherapeutic approaches being sought for its treatment. Herein, we report the generation and characterization of a monoclonal antibody, YX1-40H10, that catalyzes the photooxidation of (+)-2 into the nonpsychoactive compound benzaldehyde (14) under anaerobic conditions in the presence of riboflavin (6). Studies have revealed that the antibody facilitates the conversion of (+)-2 into 14 by binding the triplet photoexcited state of 6 in proximity to (+)-2. The antibody binds riboflavin (K(d) = 180 muM), although this was not programmed into hapten design, and the YX1-40H10-catalyzed reaction is inhibited by molecular oxygen via the presumed quenching of the photoexcited triplet state of 6. Given that this reaction is another highlight in the processing of reactive intermediates by antibodies, we speculate that this process may have future significance in vivo with programmed immunoglobulins that use flavins as cofactors to destroy selectable molecular targets under hypoxic or even anoxic conditions. PMID:17360412

  8. Immunotherapy with monoclonal antibodies in metastatic melanoma

    Microsoft Academic Search

    Thomas A. Steffens; Dean F. Bajorin; Alan N. Houghton

    1992-01-01

    Therapy for metastatic melanoma has been disappointing to date. Treatment with chemotherapy only uncommonly results in complete responses and rarely results in long-term survivors. The identification of human melanoma cell surface antigens has led to the development of an array of mouse monoclonal antibodies (MAb) for use in the diagnosis and therapy of patients with metastatic melanoma. Strategies utilizing MAbs

  9. Innovative monoclonal antibody therapies in multiple sclerosis

    Microsoft Academic Search

    Ralf A. Linker; Bernd C. Kieseier

    The recent years have witnessed great efforts in establishing new therapeutic options for multiple sclerosis (MS), especially for relapsing-remitting disease courses. In particular, the application of monoclonal antibodies provide innovative approaches allowing for blocking or depleting specific molecular targets, which are of interest in the pathogenesis of MS. While natalizumab received approval by the US Food and Drug Administration and

  10. Modulation of neurotrophin signaling by monoclonal antibodies.

    PubMed

    Rosenthal, A; Lin, J C

    2014-01-01

    The neurotrophin family is comprised of the structurally related secreted proteins nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and neurotrophine-4 (NT-4). They bind and activate the tyrosine kinase receptors Trk A, B, and C in a ligand-specific manner and additionally bind a shared p75NTR receptor. The neurotrophins were originally defined by their ability to support the survival and maturation of embryonic neurons. However, they also control important physiological functions of the adult nervous system including learning and memory, sensation, and energy homeostasis. For example, NGF/trkA signaling is critical for normal and pathological sensation of pain. Likewise, the BDNF/trkB pathway controls feeding and metabolism, and its dysfunction leads to severe obesity. Antibodies can modulate neurotrophin signaling. Thus, NGF blocking agents can attenuate pain in several animal models, and a recombinant humanized NGF blocking antibody (Tanezumab) has shown promising results in human clinical trials for osteoarthritic pain. On the other hand trkB agonist antibodies can modulate food intake and body weight in rodents and nonhuman primates. The power of monoclonal antibodies to modulate neurotrophin signaling promises to turn the rich biological insights into novel human medicines. PMID:24668485

  11. de Lange lab protocol Peptide Antibody Production

    E-print Network

    de Lange, Titia

    de Lange lab protocol Peptide Antibody Production A) Peptide BioSynthesis (http synthesis at BioSynthesis (http://www.biosyn.com, 800-227-0627). Make 24-25 aa peptide and add Cys to COOH://www.biosyn.com, 800-227-0627) B) Conjugation of peptide to KLH (Imject Maleimide Activated KLH, PIERCE=Thermo #77605

  12. Neutralizing Antibody Response to Hepatitis C Virus

    PubMed Central

    Wang, Yong; Keck, Zhen-Yong; Foung, Steven K. H.

    2011-01-01

    A critical first step in a “rational vaccine design” approach for hepatitis C virus (HCV) is to identify the most relevant mechanisms of immune protection. Emerging evidence provides support for a protective role of virus neutralizing antibodies, and the ability of the B cell response to modify the course of acute HCV infection. This has been made possible by the development of in vitro cell culture models, based on HCV retroviral pseudotype particles expressing E1E2 and infectious cell culture-derived HCV virions, and small animal models that are robust tools in studies of antibody-mediated virus neutralization. This review is focused on the immunogenic determinants on the E2 glycoprotein mediating virus neutralization and the pathways in which the virus is able to escape from immune containment. Encouraging findings from recent studies provide support for the existence of broadly neutralization antibodies that are not associated with virus escape. The identification of conserved epitopes mediating virus neutralization that are not associated with virus escape will facilitate the design of a vaccine immunogen capable of eliciting broadly neutralizing antibodies against this highly diverse virus. PMID:22163337

  13. Antiubiquitin antibody in localised and systemic scleroderma.

    PubMed Central

    Fujimoto, M; Sato, S; Ihn, H; Kikuchi, K; Tamaki, T; Tamaki, K; Takehara, K

    1996-01-01

    OBJECTIVE: To determine the presence of antiubiquitin antibody (AUbA) in localised scleroderma and systemic sclerosis, as it is frequently found in the sera of patients with systemic lupus erythematosus (SLE) and has also been shown to have a close relationship with antihistone antibodies that have an important role in scleroderma. METHODS: Serum samples from patients with localised scleroderma (n = 48) and systemic sclerosis (n = 52) were examined by enzyme linked immunosorbent assay. Twenty samples from patients with SLE, 20 from patients with dermatomyositis, and 30 samples from healthy individuals were used as controls. RESULTS: AUbA was demonstrated in 44% of patients with localised scleroderma and in 42% of those with systemic sclerosis. The presence of AUbA correlated with the presence of antihistone antibodies in both localised scleroderma and systemic sclerosis. CONCLUSIONS: AUbA is frequently present in patients with localised scleroderma and systemic sclerosis. Induction of AUbA is closely associated with that of antihistone antibodies, suggesting that ubiquitinated histone may be the target in autoimmune responses of these disorders. Images PMID:8694581

  14. Development trends for human monoclonal antibody therapeutics

    Microsoft Academic Search

    Aaron L. Nelson; Eugen Dhimolea; Janice M. Reichert

    2010-01-01

    Fully human monoclonal antibodies (mAbs) are a promising and rapidly growing category of targeted therapeutic agents. The first such agents were developed during the 1980s, but none achieved clinical or commercial success. Advances in technology to generate the molecules for study — in particular, transgenic mice and yeast or phage display — renewed interest in the development of human mAbs

  15. Losing your nerves? Maybe it's the antibodies

    Microsoft Academic Search

    Patricio T. Huerta; Paola Mina-Osorio; Czeslawa Kowal; Bruce T. Volpe; Betty Diamond

    2009-01-01

    We propose that the normal immunocompetent B cell repertoire is replete with B cells making antibodies that recognize brain antigens. Although B cells that are reactive with self antigen are normally silenced during B cell maturation, the blood–brain barrier (BBB) prevents many brain antigens from participating in this process. This enables the generation of a B cell repertoire that is

  16. Orthobunyavirus Antibodies in Humans, Yucatan Peninsula, Mexico

    PubMed Central

    Saiyasombat, Rungrat; Talavera-Aguilar, Lourdes G.; Garcia-Rejon, Julian E.; Farfan-Ale, Jose A.; Machain-Williams, Carlos; Lorońo-Pino, Maria A.

    2012-01-01

    We performed a serologic investigation to determine whether orthobunyaviruses commonly infect humans in the Yucatan Peninsula of Mexico. Orthobunyavirus-specific antibodies were detected by plaque reduction neutralization test in 146 (18%) of 823 persons tested. Further studies are needed to determine health risks for humans from this potentially deadly group of viruses. PMID:23017592

  17. Engineering receptors and antibodies for biosensors

    Microsoft Academic Search

    B Hock; M Seifert; K Kramer

    2002-01-01

    Biosensor sensitivity and selectivity depend essentially on the properties of the biorecognition elements to be used for analyte binding. Two principally different applications are considered, (1) effects monitoring with biological components as targets for bioeffective substances, among them endocrine disruptors; and (2) immunochemical analysis employing antibodies as binding proteins for a wide variety of analytes such as pesticides. Genetic engineering

  18. Rapid antibody test for fragile X syndrome

    Microsoft Academic Search

    R. Willemsen; S. Mohkamsing; B. de Vries; A. van den Ouweland; H. Galjaard; B. Oostra; D. Devys; J. L. Mandel

    1995-01-01

    Fragile X syndrome is the most common known cause of inherited mental retardation. Identification of patients and carriers of fragile X syndrome is usually done with a DNA test system but we have developed a rapid antibody to F identify fragile X patients. This non-invasive test requires only 1 or 2 drops of blood and can be used for screening

  19. Subcutaneous Administration of Monoclonal Antibodies in Oncology

    PubMed Central

    Jackisch, C.; Müller, V.; Maintz, C.; Hell, S.; Ataseven, B.

    2014-01-01

    Treatment with monoclonal antibodies (mabs) has become an established component of oncological therapy. The monoclonal antibodies available for this purpose are mainly administered intravenously in individually adapted doses according to body weight over longer treatment times. For other chronic diseases such as, for example, diabetes mellitus, the subcutaneous administration of drugs is an established therapy option. For the subcutaneous administration of larger volumes as needed for mab solutions the extracellular matrix of the subcutaneous tissue represents a problem. The co-formulation with recombinant human hyaluronidase makes the relatively pain-free administration of larger fluid volumes and thus the subcutaneous administration of monoclonal antibodies possible, as illustrated by the development of a subcutaneous formulation of trastuzumab. This constitutes a less invasive, time-optimised and flexible form of administration for patients with HER2-positive breast cancer that, with its fixed dosing possibilities, contributes to therapeutic safety. The example of trastuzumab shows that the subcutaneous administration of monoclonal antibodies can simplify oncological long-term therapy not only for the patients but also for the medical personnel. PMID:25076790

  20. Rubella antibody loss rates in Korean children.

    PubMed Central

    Ki, M.; Kim, M. H.; Choi, B. Y.; Shin, Y. J.; Park, T.

    2002-01-01

    We followed students in eight elementary schools for rubella antibody from 1993 to 1996 (602 pairs) and 1996-9 (588 pairs) in Gyeonggi Province, Korea. We tested rubella IgG and administered rubella vaccine to the children with the titres < 10 IU/ml. The loss rates of rubella IgG during the follow-up periods were 14.3 and 15.8%, respectively. Among vaccinated groups, the loss rate was 18.8%, which was significantly higher than 13.8% of the mixture of natural and vaccine-induced immunity groups. The group that had the lower preceding antibody titre had a higher loss rate of 24.8% compared to 7.2% for the group whose titre was 40 IU/ml or above. In a multivariate analysis, age and gender were not related to antibody loss rate. Under this higher rubella antibody loss rate, in order to prevent congenital rubella syndrome, the immunization for women at childbearing age appears necessary until rubella can be eliminated or controlled. PMID:12558339

  1. Monoclonal antibodies reactive with chicken interleukin-17

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In our previous study chicken interleukin -17 (chIL-17) gene was cloned from the expressed sequence tag (EST) cDNA library and initially analyzed. To further investigate biological properties of chicken IL-17, six monoclonal antibodies (mAbs) against bacterially expressed protein were produced and c...

  2. Antibody elution from red blood cells

    Microsoft Academic Search

    Harry Rubin

    1963-01-01

    An elution technique is described which gives eluates at least as potent as previously described methods. Elution can be completed in only 15 minutes. The method, a modification of that of Vos and Kelsall, is simple to carry out and requires the minimum of equipment. Studies have shown that the coating antibody is almost, but not completely, eluted.

  3. Automated Aufbau of antibody structures from given sequences using Macromoltek's SmrtMolAntibody.

    PubMed

    Berrondo, Monica; Kaufmann, Susana; Berrondo, Manuel

    2014-08-01

    This study was a part of the second antibody modeling assessment. The assessment is a blind study of the performance of multiple software programs used for antibody homology modeling. In the study, research groups were given sequences for 11 antibodies and asked to predict their corresponding structures. The results were measured using root-mean-square deviation (rmsd) between the submitted models and X-ray crystal structures. In 10 of 11 cases, the results using SmrtMolAntibody show good agreement between the submitted models and X-ray crystal structures. In the first stage, the average rmsd was 1.4 Ĺ. Average rmsd values for the framework was 1.2 Ĺ and for the H3 loop was 3.0 Ĺ. In stage two, there was a slight improvement with an rmsd for the H3 loop of 2.9 Ĺ. PMID:24777752

  4. A novel method of Multiplexed Competitive Antibody Binning for the characterization of monoclonal antibodies.

    PubMed

    Jia, Xiao-Chi; Raya, Robert; Zhang, Li; Foord, Orit; Walker, Wynn L; Gallo, Michael L; Haak-Frendscho, Mary; Green, Larry L; Davis, C Geoffrey

    2004-05-01

    We have developed a novel method of high-throughput Multiplexed Competitive Antibody Binning (MCAB). Using only a small amount of antibody and antigen, this method enables the sorting of a large, complex panel of monoclonal antibodies into different bins based on cross-competition for antigen binding. The MCAB assay builds on Luminex multiplexing bead-based technology to detect antibody competition. Because of its high sensitivity, the MCAB method is immediately applicable after identification of antigen-positive mAbs, providing information useful for advancing mAb candidates into further testing. The MCAB assay also can be used for sorting mAbs into binding groups after screening for functional activity. PMID:15183088

  5. Neuronal surface antigen antibodies in limbic encephalitis

    PubMed Central

    Graus, F; Saiz, A; Lai, M; Bruna, J; López, F; Sabater, L; Blanco, Y; Rey, M J.; Ribalta, T; Dalmau, J

    2008-01-01

    Objective: To report the frequency and type of antibodies against neuronal surface antigens (NSA-ab) in limbic encephalitis (LE). Methods: Analysis of clinical features, neuropathologic findings, and detection of NSA-ab using immunochemistry on rat tissue and neuronal cultures in a series of 45 patients with paraneoplastic (23) or idiopathic (22) LE. Results: NSA-ab were identified in 29 patients (64%; 12 paraneoplastic, 17 idiopathic). Thirteen patients had voltage-gated potassium channels (VGKC)-ab, 11 novel NSA (nNSA)-ab, and 5 NMDA receptor (NMDAR)-ab. nNSA-ab did not identify a common antigen and were more frequent in paraneoplastic than idiopathic LE (39% vs 9%; p = 0.03). When compared with VGKC-ab or NMDAR-ab, the nNSA associated more frequently with intraneuronal antibodies (11% vs 73%; p = 0.001). Of 12 patients (9 nNSA-ab, 2 VGKC-ab, 1 NMDAR-ab) with paraneoplastic LE and NSA-ab, concomitant intraneuronal antibodies occurred in 9 (75%). None of these 12 patients improved with immunotherapy. The autopsy of three of them showed neuronal loss, microgliosis, and cytotoxic T cell infiltrates in the hippocampus and amygdala. These findings were compatible with a T-cell mediated neuronal damage. In contrast, 13 of 17 (76%) patients with idiopathic LE and NSA-ab (8 VGKC-ab, 4 NMDAR-ab, 1 nNSA-ab) and 1 of 5 (20%) without antibodies had clinical improvement (p = 0.04). Conclusions: In paraneoplastic limbic encephalitis (LE), novel antibodies against neuronal surface antigens (nNSA-ab) occur frequently, coexist with antibodies against intracellular antigens, and these cases are refractory to immunotherapy. In idiopathic LE, the likelihood of improvement is significantly higher in patients with NSA-ab than in those without antibodies. GLOSSARY GAD = glutamic acid decarboxylase; LE = limbic encephalitis; NMDAR = N-methyl-D-aspartate receptor; NSA = neuronal surface antigens; nNSA = novel NSA; SCLC = small-cell lung cancer; VGKC = voltage-gated potassium channels; WBC = white blood cells. PMID:18794496

  6. Antibody responses of hens fed vitamin E and passively acquired antibodies of their chicks.

    PubMed

    Boa-Amponsem, K; Price, S E; Geraert, P A; Picard, M; Siegel, P B

    2001-01-01

    Antibody responses of hens and their progeny were studied in commercial broiler nuclear lines. Starting at 168 days of age, individually housed pullets from lines A and B were fed a 16% crude protein and 2752 kcal metabolizable energy/kg mash diet supplemented with either 10 or 300 IU/kg of vitamin E fed as dl-alpha-tocopherol acetate. Fifty-eight days later (226 days of age), 12 hens per line-vitamin E subclass were inoculated i.v. with 0.1 ml of a 2.5% suspension of sheep red blood cells (SRBC). Plasma antibody titers were measured 6, 20, 40, 54, 70, and 88 days after inoculation. Hens from both lines were artificially mated to males from line C, and progeny from eggs collected 9-15, 25-30, and 65-70 days after inoculation were tested for antibodies to SRBC. Hens were reinoculated i.v. with 0.1 ml of 0.25% SRBC 88 days after the first inoculation, and their antibody levels were measured 3, 6, and 20 days later. Eggs laid 10-13 days after reinoculation were incubated, and antibody titers of chicks were measured at hatch. Antibody response of hens to an initial inoculation of SRBC was line-diet-time after inoculation specific. In line A, titers were greater for hens fed the lower than the higher vitamin E diet, whereas diet had no effect on the antibody levels in line B. Line effects (A > B) were observed on days 6 and 20 after inoculation but not thereafter. After the second inoculation, dietary vitamin E level had no effect on antibody levels of hens within lines, whereas a between-line difference (A > B) was observed for the lower but not the higher level of dietary vitamin E. Although there was no difference between diets for antibody transferred to progeny by line B, there was a difference (lower > higher) for line A. After reinoculation of their dams, antibody titers of chicks from line A, but not line B, reached levels similar to those after the first inoculation. Antibody levels were higher for chicks at hatch than in 16-day embryos or 10 days posthatch. The results of this research suggest genetic variation in response to immune stimulation by dietary vitamin E. PMID:11332472

  7. Monoclonal antibodies to lampbrush chromosome antigens of Pleurodeles waltlii

    Microsoft Academic Search

    J. C. Lacroix; R. Azzouz; D. Boucher; C. Abbadie; C. K. Pyne; J. Charlemagne

    1985-01-01

    Germinal vesicles of oocytes from Pleurodeles waltlii were used for immunization of BALB\\/c mice to obtain hybridomas secreting monoclonal antibodies. The hybridomas were screened for reactivity of their antibodies against lampbrush chromosomes of oocytes, as revealed by indirect immunostaining. Antibodies labelling the lampbrush chromosomes were also tested on histological sections of oocytes, embryos, and larvae of Pleurodeles. Characterization of the

  8. Specific Antibody Response to Oligomannosidic Epitopes in Crohn's Disease

    Microsoft Academic Search

    B. SENDID; J. F. COLOMBEL; P. M. JACQUINOT; C. FAILLE; J. FRUIT; A. CORTOT; D. LUCIDARME; D. CAMUS

    1996-01-01

    Elevated antibody levels against the yeastSaccharomyces cerevisiaehave been reported in sera from patients with Crohn's disease and not with ulcerative colitis. The aim of the study was to identify the nature of the epitopes supporting this antibody response. Whole cells from different S. cerevisiae strains were selected in immunofluorescence assay for their ability to differentiate the antibody responses of patients

  9. The solid phase in affinity chromatography: strategies for antibody attachment

    Microsoft Academic Search

    M. Nisnevitch; M. A. Firer

    2001-01-01

    Antibodies (Ab) are commonly used in affinity chromatography (AC) as a versatile and specific means of isolating target molecules from complex mixtures. A number of procedures have been developed to immobilize antibodies on the solid matrix. Some of these methods couple the antibody via chemical groups that may be important for specific recognition of antigen, resulting in loss of functionality

  10. Artificial antibodies to corticosteroids prepared by molecular imprinting

    Microsoft Academic Search

    Olof Ramström; Lei Ye; Klaus Mosbach

    1996-01-01

    Background: Molecular imprinting can be used to prepare antibody and receptor mimics. We have previously shown that acrylic acid polymers can be imprinted to recognize a variety of small molecules. Here, we show that molecularly imprinted polymers (MIPs) can selectively recognize steroid structures.Results: Artificial antibodies mimicking the binding performance of natural anticorticosteroid antibodies have been prepared using a molecular imprinting

  11. Antibodies against Lagos bat virus in megachiroptera from West Africa.

    PubMed

    Hayman, David T S; Fooks, Anthony R; Horton, Daniel; Suu-Ire, Richard; Breed, Andrew C; Cunningham, Andrew A; Wood, James L N

    2008-06-01

    To investigate the presence of Lagos bat virus (LBV)-specific antibodies in megachiroptera from West Africa, we conducted fluorescent antibody virus neutralization tests. Neutralizing antibodies were detected in Eidolon helvum (37%), Epomophorus gambianus (3%), and Epomops buettikoferi (33%, 2/6) from Ghana. These findings confirm the presence of LBV in West Africa. PMID:18507903

  12. Antigens of infectious laryngotracheitis herpesvirus defined by monoclonal antibodies

    Microsoft Academic Search

    Jennifer J. York; S. Sonza; M. R. Brandon; K. J. Fahey

    1990-01-01

    Summary Monoclonal antibodies to glycoprotein and protein antigens of infectious laryngotracheitis virus (ILTV) were divided into five groups on the basis of their reactivity in immunofluorescence and Western blotting. Group I antibodies recognised a single band of 60 k and Group II antibodies recognised bands of 205, 160, 115, 90 and 85 k in Western blotting. In immunofluorescence both these

  13. UTILIZATION OF MONOCLONAL ANTIBODIES FOR ANTIGENIC CHARACTERIZATION OF CORONAVIRUSES

    E-print Network

    Paris-Sud XI, Université de

    UTILIZATION OF MONOCLONAL ANTIBODIES FOR ANTIGENIC CHARACTERIZATION OF CORONAVIRUSES J.F. VAUTHEROT antibodies against Bovine Enteric Coronavirus (BECV strain G110) were obtained by fusion between SP2 monoclonal antibodies was established to be anti-GP105 by immunochemical staining of viral polypeptides

  14. Aged venous thrombi: radioimmunoimaging with fibrin-specific monoclonal antibody

    SciTech Connect

    Rosebrough, S.F.; Grossman, Z.D.; McAfee, J.G.; Kudryk, B.J.; Subramanian, G.; Ritter-Hrncirik, C.A.; Witanowski, L.S.; Tillapaugh-Fay, G.; Urrutia, E.

    1987-02-01

    Radioimmunoimaging of fresh canine venous thrombi with a murine monoclonal antibody specific for human and dog fibrin has been reported. Successful imaging of canine deep venous thrombi 1, 3, and 5 days old at the time of antibody injection is reported. Images were positive in all dogs, and the uptake of fibrin-specific antibody was equivalent to that of fresh thrombi.

  15. Multiplexed measurement of serum antibodies using an array biosensor

    Microsoft Academic Search

    Maria C. Moreno-Bondi; Chris Rowe Taitt; Lisa C. Shriver-Lake; Frances S. Ligler

    2006-01-01

    The array biosensor provides the capability for simultaneously measuring titers of antibody against multiple antigens. Human antibodies against four different targets, tetanus toxin, diphtheria toxin, staphylococcal enterotoxin B (SEB) and hepatitis B, were measured simultaneously in sera from eight different donors in a single assay and titers were determined. The assays could measure amounts of bound antibody as low as

  16. Detection of Antibodies to Melanocytes in Vitiligo by Specific Immunoprecipitation

    Microsoft Academic Search

    Gail K. Naughton; Magdalena Eisinger; Jean-Claude Bystryn

    1983-01-01

    Immunoprecipitation was used to assay for antibodies to normal human melanocytes in the sera of 12 patients with common vitiligo and 12 normal individuals. The procedure is based on the specific immunoprecipitation using protein A-sepharose of antibodies binding to detergent-soluble, radioiodinated macromolecules of normal human melanocytes grown in culture. Antibodies to melanocytes were found in all 12 patients with vitiligo

  17. Monoclonal antibodies distinguish identifiable neurones in the leech

    Microsoft Academic Search

    Birgit Zipser; Ronald McKay

    1981-01-01

    Monoclonal antibodies were isolated by screening 475 hybridomas obtained from mice immunized with whole leech nerve cords. The majority (about 300) reacted with leech nervous tissue, but only about 40 made antibodies that identified single kinds or small sets of cells. Twenty of the antibodies which react with specific neurones were studied in greater detail and are described here. They

  18. FINAL REPORT. ENGINEERED ANTIBODIES FOR MONITORING OF POLYNUCLEAR AROMATIC HYDROCARBONS

    EPA Science Inventory

    This project was conducted to remove the major barrier to the timely development and use of more versatile antibody-based detection and sample cleanup methods. The main objective was to adapt combinatorial antibody library and antibody engineering methods for preparing a panel of...

  19. Heterophilic antibodies as a source of error in immunoassay

    SciTech Connect

    Witherspoon, L.; Witkin, M.; Shuler, S.; Neely, H.; Gilbert, S.

    1985-05-01

    Antibodies directed against IgG used in an immunoassay, when present in patient serum, may lead to erroneous estimates of analyte by combining with the primary antibody and effectively reducing its concentration. In two patients with anti-rabbit IgG the authors obtained apparently elevated TSH, LH, and FSH estimates using competitive, second antibody kits Dilutional parallelism could not be demonstrated using these kits. Normal TSH estimates were obtained using one of several IRMAs and a competitive assay which included rabbit IgG in the buffer. Normal LH and FSH estimates (including dilutional parallelism) were obtained using competitive assay kits which included rabbit IgG in the buffer. The authors were not able to repair the antibody-limited kits by adding rabbit IgG, as the second antibody concentration was inadequate to precipitate the added IgG Heterophilic antibody directed against the assay antibody presents a significant potential problem in immunoassay. This problem is most pronounced in antibody limited systems and may be avoided by the addition of same-species IgG. The laboratory user may not be able to make this addition unless the separation step is reoptomized. Antibody excess (IRMA) systems are effected if the offending antibody is present in concentrations sufficient to saturate the extracting antibody. Practically, a heterophilic antibody must be suspected by demonstrating nonparallelism. This potential problem should be more widely appreciated.

  20. Antibody Overview 1-3 Introduction to Antibody Production, Purification and Modification 1

    E-print Network

    Lebendiker, Mario

    Fragments 42 Fragmentation of IgG 43 Fragmentation of IgM 48 Antibody Labeling 50-69 Overview 50 Enzyme Labeling 51 Biotin Labeling 57 Fluorescent Labeling 63 Iodine Labeling 67 Contents #12;This ability (ELISA). Determining the class (e.g., IgG vs. IgM) and subclass (e.g., IgG1 vs. IgG2a) of an antibody

  1. Development of antibody arrays for monoclonal antibody Higher Order Structure analysis.

    PubMed

    Wang, Xing; Li, Qing; Davies, Michael

    2013-01-01

    Antibody arrays were developed to probe a monoclonal antibody's three-dimensional structure (3-D structure). Peptides with overlapping regions were designed to cover the whole mAb light chain and heavy chain, respectively, and used to generate polyclonal antibodies after the conjugation of the peptides to a carrier protein, KLH. It was shown that good peptide specificity was achieved from the antibodies generated. Using more than 30 different polyclonal antibodies to measure the surface epitope distribution, it was shown that the mAb antibody array can detect epitope exposure as low as 0.1% of defined mAb populations. This ELISA-based analysis of mAb epitope exposure can be considered as a measurement of "conformational impurity" in biologics development, similar to the analysis of other product-related impurities such as different forms of glycosylation, deamidation, and oxidation. This analysis of "conformational impurity" could provide valuable information on the mAb conformational comparability for biosimilar mAbs as well as novel mAbs, especially in the area of protein immunogenicity. Furthermore, stability studies indicated that there are several conformational "hot spots" in many mAbs tested, especially in the hinge region. This antibody array technology can be used for novel mAb Higher Order Structure (HOS) analysis during process and formulation development. Another important area of application is for biosimilar mAb development where the innovator molecule and biosimilar molecule could be compared based on their systemic "fingerprint" from the 30 plus antibodies. PMID:23970865

  2. Transplacental transfer of immune antibodies in the mouse demonstrated by antibody labeled in vivo with tritium 

    E-print Network

    McKinney, Hubert Eugene

    1971-01-01

    of the requirement for the degree of MASTER OF SCIENCE August 1971 Major Sub] ect: Laboratory Animal Medicine TRAYSPLACENTAL TRANSFER OF IMMUNE ANTIBODIES IN THE MOUSE DEMONSTRATED BY ANTIBODY LABELED IN VIVO WITH TRITIUM A Thesis by HUBERT EUGENE MCKINNEY.... Pooled sera of stock mice, from which the animals used in this study originated, did not neutralize the immuniz- ing virus in any dilution. Serum collected from passively immunized mice 6 or more days after immunization did not neutralize the immuniz...

  3. Panel reactive antibody positivity and associated HLA antibodies in Turkish renal transplant candidates.

    PubMed

    Ozdemir, Fatma Nurhan; Sezer, Siren; Akcay, Ali; Arat, Zubeyde; Turan, Munire; Gulmus, Sale; Kulah, Eyup; Haberal, Mehmet

    2004-01-01

    Pre- and post-renal transplantation panel reactive antibody (PRA) screening is associated with increased incidence of hyperacute or acute graft rejection and graft loss. This study was designed to find any relationship PRA sensitization and associated human leukocyte antigen (HLA)-specific antibodies in Turkish renal transplant candidates. We included 340 patients who were in the renal transplantation waiting list in the study. We determined PRA sensitization ratio and the associated anti-HLA IgG antibody distribution of the patient group. The PRA testing was currently performed and levels above 30% were accepted to be positive. The PRA class I positivity was determined in 24 (7%) and class II in 34 (10%) of the patients. The most frequent HLA antibodies for class I were B56, A2, A34, A1, A23, A24 and B61; and for class II were DR11, DR14, DQ7, DR10, DQ5, DR1 and DR7, respectively. From these, the increase of the numbers of anti-HLA class II antibodies was significantly correlated with the increase of PRA sensitization ratio. In conclusion, the identification of the associated HLA-specific antibodies and correlation with the Turkish population HLA antigen distribution will identify the high-risk patients who are candidates for transplantation. PMID:14967317

  4. [The isolation and testing of syngeneic anti-idiotypic antibodies against antimycobacterial monoclonal antibodies].

    PubMed

    Avdienko, V G; Kramnik, I B; Apt, A S; Litvinov, V I

    1993-01-01

    Fifteen monoclonal antibodies (MAb) were obtained to cell walls (CW) of M. bovinus-8, two of them were limited specific against human and bovine mycobacteria. One MAb reacted in immunoblotting with a protein having molecular mass of 19.1 kDa. It was also found that all MAb bind with the antigen determinants of mycobacterial proteins. The competitive enzyme immunoassay (EIA) helped reveal that the antigenic determinants "recognized" by two MAb are located in the same area despite the fact that one reacted in immunoblotting with a denatured protein and the other "recognized" only a native antigen in EIA. The syngeneic antiidiotypic (anti-ID) immune response was induced by these MAb in BALB/c mice. The EIA showed the binding of anti-ID-antibodies isolated from mice serum both MAb inducing their synthesis and to antimycobacterial serum antibodies of caws with tuberculosis. Data suggesting a similarity existing between the mycobacterial antigen and anti-ID-antibodies were also obtained in the blast transformation reaction: in M. bovinus antigen stimulation of 8 mice lymph node cells sensitized by anti-ID-antibodies and in the reverse situation when the cells sensitive to KC M. bovinus-8 proliferated in response to stimulation by anti-ID-antibodies. PMID:7687055

  5. Antibody zymography: a novel adaptation of zymography to determine the protease-neutralising potential of specific antibodies and snake antivenoms

    Microsoft Academic Search

    S. S. Hasson; R. D. G. Theakston; R. A. Harrison

    2004-01-01

    A common problem in the development of antibody-based therapeutics is the selection, usually from a large population, of specific antibodies with the desired function. One of our research objectives is to identify antibodies capable of neutralising the most important haemorrhagic and haemostasis-disruptive proteases from viper venom. Here, we describe a modification of conventional gelatin-zymography that permits the identification of antibodies

  6. Development of Tetravalent, Bispecific CCR5 Antibodies with Antiviral Activity against CCR5 Monoclonal Antibody-Resistant HIV-1 Strains?

    PubMed Central

    Schanzer, Jürgen; Jekle, Andreas; Nezu, Junichi; Lochner, Adriane; Croasdale, Rebecca; Dioszegi, Marianna; Zhang, Jun; Hoffmann, Eike; Dormeyer, Wilma; Stracke, Jan; Schäfer, Wolfgang; Ji, Changhua; Heilek, Gabrielle; Cammack, Nick; Brandt, Michael; Umana, Pablo; Brinkmann, Ulrich

    2011-01-01

    In this study, we describe novel tetravalent, bispecific antibody derivatives that bind two different epitopes on the HIV coreceptor CCR5. The basic protein formats that we applied were derived from Morrison-type bispecific antibodies: whole IgGs to which we connected single-chain antibodies (scFvs) via (Gly4Ser)n sequences at either the C or N terminus of the light chain or heavy chain. By design optimization, including disulfide stabilization of scFvs or introduction of 30-amino-acid linkers, stable molecules could be obtained in amounts that were within the same range as or no less than 4-fold lower than those observed with monoclonal antibodies in transient expression assays. In contrast to monospecific CCR5 antibodies, bispecific antibody derivatives block two alternative docking sites of CCR5-tropic HIV strains on the CCR5 coreceptor. Consequently, these molecules showed 18- to 57-fold increased antiviral activities compared to the parent antibodies. Most importantly, one prototypic tetravalent CCR5 antibody had antiviral activity against virus strains resistant to the single parental antibodies. In summary, physical linkage of two CCR5 antibodies targeting different epitopes on the HIV coreceptor CCR5 resulted in tetravalent, bispecific antibodies with enhanced antiviral potency against wild-type and CCR5 antibody-resistant HIV-1 strains. PMID:21300827

  7. Regeneration of ethyl parathion antibodies for repeated use in immunosensor: a study on dissociation of antigens from antibodies

    Microsoft Academic Search

    V. B. Kandimalla; N. S. Neeta; N. G. Karanth; M. S. Thakur; K. R. Roshini; B. E. A. Rani; A. Pasha

    2004-01-01

    Reliable analysis using an immunosensor strongly depends on the specificity, activity, and sensitivity of the antibody. Immobilization of antibody on the solid matrix enables its repeated use, for which it is required to dissociate the antigens and antigen–enzyme conjugate from the immobilized antibody matrix after each use and while doing so, a maximum retention of activity and specificity are crucial

  8. Paradoxical suppression of poly-specific broadly neutralizing antibodies in the presence of strain-specific neutralizing antibodies following

    E-print Network

    De Leenheer, Patrick

    studies have shown the existence of monoclonal antibodies that exhibit broad crossclade neutralizing. Antibodies directed against HIV structural proteins are detected in the body within a few weeks following, however, neutralize the virus, which escapes recognition by ensuing reduced accessibility to antibody

  9. Antibody response to Epstein-Barr virus in infectious mononucleosis.

    PubMed Central

    Nikoskelainen, J; Hänninen, P

    1975-01-01

    Altogether 171 serum specimens from 58 patients with heterophil antibody-positive infectious monomucleosis were studied for antibody response to Epstein-Barr virus (EBV). The sera were tested for fluorescent immunoglobulin G (IgG) and IgM gel-precipitating (GP) and complement-fixing (CF) antibodies to EBV. All 58 patients had IgG and IgM antibodies to EBV. Both IgG and IgM antibodies developed rapidly; the IgM antibodies disappeared within 8 to 10 weeks, whereas the IgG antibodies remained at an almost constant level. The development of IgG antibodies was so rapid that a fourfold or greater rise in titers was noted only in 22% of the patients. Both GP and CF antibodies to EBV (crude P3HR-1 Burkitt cell antigen) developed slowly; the mean titers kept rising for more than 12 weeks. The micro GP technique seemed to be more sensitive than the CF method, because 86% of the patients with infectious mononucleosis had GP antibodies compared with 72% having CF antibodies. In patients with infectious mononucleosis, a seroconversion or significant rise in GP antibodies was noted in 57%, whereas only 19% had a similar change in CF antibodies. The most promising of these antibody assays in the diagnosis of recent infections was the EBV-specific IgM antibody technique, which enables one to make the diagnosis on the basis of only one serum specimen. In cases where the acute-phase serum specimen is missing, the diagnosis can be made later by using the GP and CF techniques. Images PMID:163790

  10. Pitfalls of formalin fixation for determination of antineutrophil cytoplasmic antibodies.

    PubMed Central

    Chowdhury, S M; Broomhead, V; Spickett, G P; Wilkinson, R

    1999-01-01

    Sera can produce nuclear or perinuclear immunofluorescence staining in neutrophils which may be caused by antibodies with differing antigenic specificities. These include perinuclear antineutrophil cytoplasmic antibodies (P-ANCA), granulocyte specific antinuclear antibody (GS-ANA), and antinuclear antibody (ANA). There is controversy over the value of formalin fixation of neutrophils in differentiating antibodies giving selective or preferential reaction with the nuclear or perinuclear area of neutrophils. In a comparative study of 77 sera, formalin fixation caused inconsistency, nonspecific effects, and false positivity owing to enhanced fluorescence. If formalin fixed neutrophils are used in the routine diagnostic laboratory, this will add confusion to the interpretation of the ANCA assay. PMID:10562820

  11. Strategies for enhancing antibody delivery to the brain.

    PubMed

    Frank, Richard T; Aboody, Karen S; Najbauer, Joseph

    2011-12-01

    Antibodies and antibody conjugates have emerged as important tools for cancer therapy. However, a major therapeutic challenge for the use of antibodies is their inability to cross the blood-brain barrier (BBB) to reach tumors localized in the central nervous system (CNS). Multiple methods have been developed to enhance antibody delivery to the CNS, including direct injection, mechanical or biochemical disruption of the BBB, conjugation to a 'molecular Trojan horse', cationization, encapsulation in nanoparticles and liposomes, and more recently, stem cell-mediated antibody delivery. In this review, we discuss each of these approaches, highlighting their successes and the obstacles that remain to be overcome. PMID:21767610

  12. Epitope mapping of antibodies by mass spectroscopy: a case study.

    PubMed

    Obungu, Victor H; Gelfanova, Valentina; Huang, Lihua

    2013-01-01

    Epitope mapping of antibodies is the identification and characterization of binding sites of monoclonal antibodies (mAbs) on target antigens. This knowledge can be useful in generating novel antibodies to a particular target as well as elucidating an antibody mechanism of action. Several techniques are available to identify antibody epitopes among which are preliminary and simple ones like sequence homology analysis ELISA and Western blotting. However, the more widely used robust methods typically involve the use of mass spectrometry to fully analyze and interpret the data and accurately identify the binding site. Such methods include epitope extortion/excision, hydrogen deuterium exchange. PMID:23475727

  13. [Separation strategy of affinity chromatography for mixed antibodies].

    PubMed

    Zhou, Yuefang; Zhang, Yan; Luo, Jian; Kang, Limei; Chen, Yi; Shi, Hong; Meng, Qingxiong; Su, Zhiguo

    2013-10-01

    A mammary gland bioreactor can efficiently express human recombinant monoclonal antibody. However, the target products are similar to the bovine antibody in the raw emulsion material in properties and structures. Thus it is difficult to achieve effective separation of the target products. In this work, the species differences between bovine antibody and recombinant human antibody were analyzed and a new separation strategy was raised based on it. We employed two kinds of affinity chromatography to separate these two antibodies from each other and studied the effect of elution mode upon separation. The results demonstrated that Protein A affinity chromatography could get hybrid antibodies using gradient elution mode, but hardly separate the recombinant human antibody and bovine antibody from each other. In contrast, the combination of Protein A affinity chromatography and displacement chromatography could separate the hybrid antibodies effectively and finally give recombinant human IgG (rHGG) product with the purity of 95% and the yield of more than 95%. Immuno-affinity chromatography could also effectively purify recombinant monoclonal antibodies and owned better generality, which could be used in purification of recombinant antibody expressed by any animal mammary gland. - PMID:24432640

  14. Antibody engineering for increased potency, breadth and half-life

    PubMed Central

    Sievers, Stuart A.; Scharf, Louise; West, Anthony P.; Bjorkman, Pamela J.

    2015-01-01

    Purpose of review This review highlights recent developments in HIV-1 antibody engineering and discusses the effects of increased polyreactivity on serum half-lives of engineered antibodies. Recent findings Recent studies have uncovered a wealth of information about the relationship between the sequences and efficacies of anti-HIV-1 antibodies through a combination of bioinformatics, structural characterization and in vivo studies. This knowledge has stimulated efforts to enhance antibody breadth and potency for therapeutic use. Although some engineered antibodies have shown increased polyreactivity and short half-lives, promising efforts are circumventing these problems. Summary Antibodies are desirable as therapeutics due to their ability to recognize targets with both specificity and high affinity. Furthermore, the ability of antibodies to stimulate Fc-mediated effector functions can increase their utility. Thus, mAbs have become central to strategies for the treatment of various diseases. Using both targeted and library-based approaches, antibodies can be engineered to improve their therapeutic properties. This article will discuss recent antibody engineering efforts to improve the breadth and potency of anti-HIV-1 antibodies. The polyreactivity of engineered HIV-1 bNAbs and the effect on serum half-life will be explored along with strategies to overcome problems introduced by engineering antibodies. Finally, advances in creating bispecific anti-HIV-1 reagents are discussed. PMID:25760931

  15. Studies on production of anticollagen antibodies in silicosis

    SciTech Connect

    Nagaoka, Tadasu; Tabata, Masaji; Kobayashi, Kenichi; Okada, Akira (Kanazawa Univ. (Japan))

    1993-01-01

    Silicosis is characterized by pulmonary fibrotic changes which consist primarily of an increase in collagen. In this study, anticollagen antibodies in the serum of 134 silicosis patients versus 40 normal subjects were examined and their relationship with immunoglobulin, autoantibodies, and procollagen III peptide (PIIIP) was investigated by enzyme-linked immunosorbent assay (ELISA). The mean levels of antihuman type I collagen (HI) and anti-human type Ill collagen (HIII) antibodies were significantly higher in the silicosis patients versus the normal subjects (P < 0.001). However, no differences were observed in the mean levels of anti-human type IV collagen (HIV) antibodies in the silicosis patients versus the normal subjects. Anticollagen antibodies in the sera of silicosis patients appear to be formed at an early stage of the disease. We observed a correlation between anticollagen antibodies and immunoglobulin. There was a tendency toward high values of anticollagen antibodies in the sera of patients positive for antinuclear antibodies (ANA) and rheumatoid factor (RF), both of which are autoantibodies. However, no correlation was observed between serum PIIIP and anticollagen antibodies. These observations suggest that, in silicosis, there is a relationship between anticollagen antibodies and immunoglobulins, as well as between anticollagen antibodies and autoantibodies. Measurement of anticollagen antibodies in the sera of silicosis patients offers a useful index for evaluating the prognosis of pulmonary fibrosis and autoimmune abnormality in silicosis. 49 refs. 6 figs., 6 tabs.

  16. Discovery of internalizing antibodies to tumor antigens from phage libraries

    PubMed Central

    Zhou, Yu; Marks, James D

    2014-01-01

    Phage antibody technology can be used to generate human antibodies to essentially any antigen. Many therapeutic target antigens are cell surface receptors, which can be challenging targets for antibody generation. In addition, for many therapeutic applications, one needs antibodies that not only bind the cell surface receptor but that also are internalized into the cell upon binding. This allows use of the antibody to deliver a range of payloads into the cell to achieve a therapeutic effect. In this chapter we describe how human phage antibody libraries can be selected directly on tumor cell lines to generate antibodies that bind cell surface receptors and which upon binding are rapidly internalized into the cell. Specific protocols show how to: 1) directly select cell binding and internalizing antibodies from human phage antibody libraries; 2) screen the phage antibodies in a high throughput flow cytometry assay for binding to the tumor cell line used for selection; 3) identify the antigen bound by the phage antibody using immunoprecipitation and mass spectrometry; and 4) direct cell binding and internalizing selections to a specific tumor antigen by sequential selection on a tumor cell line followed by selection on yeast displaying the target tumor antigen on the yeast surface. PMID:22208981

  17. Following single antibody binding to purple membranes in real time

    PubMed Central

    Kienberger, Ferry; Mueller, Harald; Pastushenko, Vassili; Hinterdorfer, Peter

    2004-01-01

    Antibody binding to surface antigens in membranes is the primary event in the specific immune defence of vertebrates. Here we used force microscopy to study the dynamics of antibody recognition of mutant purple membranes from Halobacterium salinarum containing a genetically appended anti-Sendai recognition epitope. Ligation of individual anti-Sendai antibodies to their antigenic epitopes was observed over time. Their increase in number within a small selected area revealed an apparent kinetic on-rate. The membrane-bound antibodies showed many different conformations that ranged from globular to V- and Y-like shapes. The maximum distance of two Fab fragments of the same antibody was observed to be ?18 nm, indicating an overall strong intrinsic flexibility of the antibody hinge region. Fab fragments of bound anti-Sendai antibodies were allocated to antigenic sites of the purple membrane, allowing the identification and localization of individual recognition epitopes on the surface of purple membranes. PMID:15143343

  18. Antibody vs. HIV in a clash of evolutionary titans.

    PubMed

    Burton, Dennis R; Stanfield, Robyn L; Wilson, Ian A

    2005-10-18

    HIV has evolved many strategies to avoid neutralizing antibody responses, particularly to conserved regions on the external glycoprotein spikes of the virus. Nevertheless, a small number of antibodies have been evolved by the human immune system to recognize conserved parts of the glycoproteins, and therefore, have broadly neutralizing cross-strain activities. These antibodies constitute important tools in the quest to design immunogens that can elicit broadly neutralizing antibodies in humans and hence contribute to an effective HIV vaccine. Crystallographic analyses of the antibodies, in many cases in an antigen-complexed form, have revealed novel and, in some instances, remarkable structural adaptations to attain virus recognition. Antibodies, like HIV, can evolve relatively rapidly through mutation and selection. It seems that the structures of these broadly neutralizing antibodies bear witness to a heroic struggle between two titans of rapid evolution. PMID:16219699

  19. Antibody-mediated cofactor-driven reactions

    DOEpatents

    Schultz, Peter G. (Oakland, CA)

    1993-01-01

    Chemical reactions capable of being rate-enhanced by auxiliary species which interact with the reactants but do not become chemically bound to them in the formation of the final product are performed in the presence of antibodies which promote the reactions. The antibodies contain regions within their antigen binding sites which recognize the auxiliary species in a conformation which promotes the reaction. The antigen binding site frequently recognizes a particular transition state complex or other high energy complex along the reaction coordinate, thereby promoting the progress of the reaction along the desired route as opposed to other less favorable routes. Various classes of reaction together with appropriate antigen binding site specificities tailored for each are disclosed.

  20. Antibody-mediated cofactor-driven reactions

    SciTech Connect

    Schultz, P.G.

    1993-06-15

    Chemical reactions capable of being rate-enhanced by auxiliary species which interact with the reactants but do not become chemically bound to them in the formation of the final product are performed in the presence of antibodies which promote the reactions. The antibodies contain regions within their antigen binding sites which recognize the auxiliary species in a conformation which promotes the reaction. The antigen binding site frequently recognizes a particular transition state complex or other high energy complex along the reaction coordinate, thereby promoting the progress of the reaction along the desired route as opposed to other less favorable routes. Various classes of reactions together with appropriate antigen binding site specificities tailored for each are disclosed.

  1. Antibody Production and Catabolism in Uraemia

    PubMed Central

    Souhami, R. L.

    1973-01-01

    A method for the production of chronic uraemia in mice is described. Production of humoral antibody has been studied in uraemic mice in response to a primary and secondary challenge with sheep red cells and in response to intraperitoneal and subcutaneous bovine serum albumin (BSA) in complete Freund's adjuvant. In uraemic animals the response to sheep red cells was slightly reduced compared with controls in both the primary and secondary responses. Similarly, the responses to intraperitoneal BSA and subcutaneous BSA in adjuvant was slightly reduced in the uraemic group. The catabolic rate of purified [125I] mouse IgG was normal in uraemic animals, indicating that the lower titre of humoral antibody was not due to increased catabolism of immunoglobulin in uraemia. PMID:4726091

  2. Monoclonal antibody to growth hormone receptors

    SciTech Connect

    Simpson, J.S.A.; Friesen, H.G.

    1985-01-01

    Using hybridoma technology, monoclonal antibodies to growth hormone receptors can be prepared in large quantities with only a few micrograms of purified antigen by in vitro immunization or by immunization of larger quantities of less pure material. The discussion centers on areas most pertinent to receptors such as receptor preparation, immunization procedure, fusion method, screening assay, and identification of the immunoglobulin class. The specificity of the antibody for growth hormone receptor was examined by testing the ability of the ascitic fluid to inhibit binding of (/sup 125/I) growth hormone to the prolactin receptors on rabbit mammary gland membranes and the inhibition of /sup 125/I-labelled rat growth hormone binding to rabbit liver membrane.

  3. Efficient antibody-catalyzed oxygenation reaction

    SciTech Connect

    Hsieh, L.C.; Stephans, J.C.; Schultz, P.G. (Univ. of California, Berkeley, CA (United States))

    1994-03-09

    Biological oxygen-transfer reactions are essential for the biosynthesis of steroids and neurotransmitters, the degradation of endogenous substances, and the detoxification of xenobiotics. The monooxygenase enzymes responsible for these transformations require biological cofactors such as flavin, heme and non-heme iron, copper, or pterin and typically utilize NADPH for cofactor regeneration. We now report an antibody-catalyzed sulfide oxygenation reaction mediated by the chemical cofactor sodium periodate, with turnover numbers similar to those of the corresponding enzymatic reactions. Sodium periodate NaIO[sub 4]O was chosen as the oxidant, since sulfoxide formation occurs under mild aqueous conditions with minimal overoxidation to the sulfone. Furthermore, compared to the flavin and heme cofactors required by the monooxygenase enzymes, NaIO[sub 4] is very inexpensive, obviating the need for cofactor recycling. Overall, these results raise the possibility of using antibodies as catalysts for regio- and stereoselective sulfide oxidations. 18 refs., 1 fig.

  4. Fluorometric assay for red blood cell antibodies

    SciTech Connect

    Schreiber, A.B.; Lambermont, M.; Strosberg, A.D.; Wybran, J.

    1981-03-01

    A fluorometric assay is described for the detection of red blood cell antibodies. The assay reveals as little as 600 molecules of bound, fluoroesceinated rabbit anti-human IgG antibodies per erythrocyte. Eleven patients with possible autoimmune erythrocyte disorder and negative direct antiglobulin test were studied by the fluorometric assay. The outcome of the fluorometric assay was compared with that of the human allogeneic rosette test. Results obtained by the two methods were in complete agreement. Five of the patients were shown to possess unexpectedly high levels of erythrocyte-bound IgG in spite of a negative, direct antiglobulin test. These findings and the validity of the fluorometric assay are discussed.

  5. Production of antibodies in transgenic plants

    Microsoft Academic Search

    Andrew Hiatt; Robert Caffferkey; Katherine Bowdish

    1989-01-01

    COMPLEMENTARY DNAs derived from a mouse hybridoma messenger RNA were used to transform tobacco leaf segments followed by regeneration of mature plants. Plants expressing single gamma or kappa immunoglobulin chains were crossed to yield progeny in which both chains were expressed simultaneously. A functional antibody accumulated to 1.3% of total leaf protein in plants expressing full-length cDNAs containing leader sequences.

  6. The cancer recognition (CARE) antibody test

    Microsoft Academic Search

    Jerry T. Thornthwaite; Emily C. McDuffee; Robert B. Harris; Julie R. Secor McVoy

    2004-01-01

    The cancer recognition (CARE) antibody (Ab) test is a serologic assay for a specific IgM that is elevated in cancer patients. All tests are measured using an indirect enzyme-linked immunosorbent assay (ELISA) of human serum. The target polypeptide in the CARE Ab test is the IgM binding epitope (LT-11) of the CARE antigen (Ag) consisting of a 16 mer structure

  7. Human Monoclonal Antibodies from Transgenic Mice

    Microsoft Academic Search

    N. Lonberg

    Since the 1986 regulatory approval of muromonomab-CD3, a mouse monoclonal antibody (MAb) directed against the T cell CD3?\\u000a antigen, MAbs have become an increasingly important class of therapeutic compounds in a variety of disease areas ranging from\\u000a cancer and autoimmune indications to infectious and cardiac diseases. However, the pathway to the present acceptance of therapeutic\\u000a MAbs within the pharmaceutical industry

  8. West Nile Virus Antibodies in Wild

    E-print Network

    Figuerola, Jordi

    medium (5), after which 50 µL of a suspension (2 × 104 cells/mL) of Vero cells plus fetal calf serum was then centrifuged (10 min at 6,000 rpm), and the serum was stored in liquid nitrogen and transported to a deep previously (5). We used USUV as a control for WNV antibody specificity. Serum samples were inactivated at 56

  9. Antibody-mediated Hsp70 protein therapy

    Microsoft Academic Search

    James E. Hansen; William Sohn; Charles Kim; Sophia S. Chang; Natalie C. Huang; Donaldson G. Santos; Grace Chan; Richard H. Weisbart; Robert N. Nishimura

    2006-01-01

    Intracellular Hsp70 provides cytoprotection against a variety of stressful stimuli, and an effective means of increasing intracellular Hsp70 levels could prove beneficial in the prevention and treatment of a variety of human diseases. A novel protein transduction domain consisting of the single chain Fv fragment of an anti-DNA antibody known to penetrate into living cells and tissues, mAb 3E10, has

  10. Antibody-Based Vascular Tumor Targeting

    Microsoft Academic Search

    Christoph Schliemann; Dario Neri

    \\u000a The inhibition of angiogenesis represents a major step toward a more selective and better-tolerated therapy of cancer. An\\u000a alternative way to take advantage of a tumor’s absolute dependence on a functional neovasculature is illustrated by the strategy\\u000a of “antibody-based vascular tumor targeting.” This technology aims at the selective delivery of bioactive molecules to the\\u000a tumor site by their conjugation to

  11. Antineutrophil Cytoplasmic Antibodies, Autoimmune Neutropenia, and Vasculitis

    PubMed Central

    Grayson, Peter C.; Sloan, J. Mark; Niles, John L.; Monach, Paul A.; Merkel, Peter A.

    2011-01-01

    Objectives Reports of an association between antineutrophil cytoplasmic antibodies (ANCA) and autoimmune neutropenia have rarely included cases of proven vasculitis. A case of ANCA-associated vasculitis (AAV) with recurrent neutropenia is described and relevant literature on the association between ANCA, neutropenia, and vasculitis is reviewed. Methods Longitudinal clinical assessments and laboratory findings are described in a patient with AAV and recurrent episodes of profound neutropenia from December 2008 – October 2010. A PubMed database search of the medical literature was performed for papers published from 1960 through October 2010 to identify all reported cases of ANCA and neutropenia. Results A 49 year-old man developed recurrent neutropenia, periodic fevers, arthritis, biopsy-proven cutaneous vasculitis, sensorineural hearing loss, epididymitis, and positive tests for ANCA with specificity for antibodies to both proteinase 3 and myeloperoxidase. Antineutrophil membrane antibodies were detected during an acute neutropenic phase and were not detectable in a post-recovery sample, whereas ANCA titers did not seem to correlate with neutropenia. An association between ANCA and neutropenia has been reported in 74 cases from 24 studies in the context of drug/toxin exposure, underlying autoimmune disease, or chronic neutropenia without underlying autoimmune disease. In these cases, the presence of atypical ANCA patterns and other antibodies were common; however, vasculitis was uncommon and when it occurred was usually limited to the skin and in cases of underlying toxin exposure. Conclusions ANCA is associated with autoimmune neutropenia, but systemic vasculitis rarely occurs in association with ANCA and neutropenia. The interaction between neutrophils and ANCA may provide insight into understanding both autoimmune neutropenia and AAV. PMID:21507463

  12. Anti-glomerular basement membrane antibodies.

    PubMed

    Silvarińo, Ricardo; Noboa, Oscar; Cervera, Ricard

    2014-11-01

    Basement membranes form an anatomic barrier that contains connective tissue. They are composed of type IV collagen, laminin and proteoglycans. Anti-basement membrane antibodies bind to the non-collagen site of the ?3 chain of type IV collagen. A group of renal diseases, pulmonary diseases and perhaps others affecting different organs have long been associated with the presence of antibodies directed against glomerular basement membrane (GBM), alveolar basement membrane and tubular basement membrane. Goodpasture disease has a frequency of 0.5 to 1 case by million/year, and is responsible for up to 20% of crescentic glomerulonephritis in renal biopsy. It has been associated with genetic and immune abnormalities and there are usually environmental triggers preceding clinical onset. Renal disease can occur isolated or in association with pulmonary hemorrhage. In general, renal disease has a rapid progression that determines severe compromise, with rare spontaneous resolution. The diagnosis of Goodpasture disease requires the presence of the anti-GBM antibody, either in circulation or in renal tissue. The prognosis of non-treated patients is poor. The standard of care is plasma exchange combined with prednisone and cyclophosphamide. Anti-GBM antibody levels must be monitored frequently until their disappearance, and then every 6 months to confirm sustained remission in the absence of clinical signs of recurrence. Prognosis of the disease is strongly associated with its initial presentation. Survival rates are related to the degree of renal compromise at onset of the disease. Recurrence of the disease post-transplantation is low. PMID:25558706

  13. Polyclonal antibody to soman-tyrosine

    PubMed Central

    Li, Bin; Duysen, Ellen G.; Froment, Marie-Thérčse; Masson, Patrick; Nachon, Florian; Jiang, Wei; Schopfer, Lawrence M.; Thiele, Geoffrey M.; Klassen, Lynell W.; Cashman, John; Williams, Gareth R.; Lockridge, Oksana

    2013-01-01

    Soman forms a stable, covalent bond with tyrosine 411 of human albumin, with tyrosines 257 and 593 in human transferrin, and with tyrosine in many other proteins. The pinacolyl group of soman is retained, suggesting that pinacolyl methylphosphonate bound to tyrosine could generate specific antibodies. Tyrosine in the pentapeptide RYGRK was covalently modified with soman simply by adding soman to the peptide. The phosphonylated-peptide was linked to keyhole limpet hemocyanin, and the conjugate was injected into rabbits. The polyclonal antiserum recognized soman-labeled human albumin, soman-mouse albumin, and soman human transferrin, but not non-phosphonylated control proteins. The soman-labeled tyrosines in these proteins are surrounded by different amino acid sequences, suggesting that the polyclonal recognizes soman-tyrosine independent of the amino acid sequence. Antiserum obtained after 4 antigen injections over a period of 18 weeks was tested in a competition ELISA where it had an IC50 of 10?11 M. The limit of detection on Western blots was 0.01 ?g (15 picomoles) of soman-labeled albumin. In conclusion, a high-affinity, polyclonal antibody that specifically recognizes soman adducts on tyrosine in a variety of proteins has been produced. Such an antibody could be useful for identifying secondary targets of soman toxicity. PMID:23469927

  14. Antibody-Based Therapies in Multiple Myeloma

    PubMed Central

    Tai, Yu-Tzu; Anderson, Kenneth C.

    2011-01-01

    The unmet need for improved multiple myeloma (MM) therapy has stimulated clinical development of monoclonal antibodies (mAbs) targeting either MM cells or cells of the bone marrow (BM) microenvironment. In contrast to small-molecule inhibitors, therapeutic mAbs present the potential to specifically target tumor cells and directly induce an immune response to lyse tumor cells. Unique immune-effector mechanisms are only triggered by therapeutic mAbs but not by small molecule targeting agents. Although therapeutic murine mAbs or chimeric mAbs can cause immunogenicity, the advancement of genetic recombination for humanizing rodent mAbs has allowed large-scale production and designation of mAbs with better affinities, efficient selection, decreasing immunogenicity, and improved effector functions. These advancements of antibody engineering technologies have largely overcome the critical obstacle of antibody immunogenicity and enabled the development and subsequent Food and Drug Administration (FDA) approval of therapeutic Abs for cancer and other diseases. PMID:22046572

  15. Constant Domain-regulated Antibody Catalysis*

    PubMed Central

    Sapparapu, Gopal; Planque, Stephanie; Mitsuda, Yukie; McLean, Gary; Nishiyama, Yasuhiro; Paul, Sudhir

    2012-01-01

    Some antibodies contain variable (V) domain catalytic sites. We report the superior amide and peptide bond-hydrolyzing activity of the same heavy and light chain V domains expressed in the IgM constant domain scaffold compared with the IgG scaffold. The superior catalytic activity of recombinant IgM was evident using two substrates, a small model peptide that is hydrolyzed without involvement of high affinity epitope binding, and HIV gp120, which is recognized specifically by noncovalent means prior to the hydrolytic reaction. The catalytic activity was inhibited by an electrophilic phosphonate diester, consistent with a nucleophilic catalytic mechanism. All 13 monoclonal IgMs tested displayed robust hydrolytic activities varying over a 91-fold range, consistent with expression of the catalytic functions at distinct levels by different V domains. The catalytic activity of polyclonal IgM was superior to polyclonal IgG from the same sera, indicating that on average IgMs express the catalytic function at levels greater than IgGs. The findings indicate a favorable effect of the remote IgM constant domain scaffold on the integrity of the V-domain catalytic site and provide a structural basis for conceiving antibody catalysis as a first line immune function expressed at high levels prior to development of mature IgG class antibodies. PMID:22948159

  16. Evaluation of Surface Chemistries for Antibody Microarrays

    SciTech Connect

    Seurynck-Servoss, Shannon L.; White, Amanda M.; Baird, Cheryl L.; Rodland, Karin D.; Zangar, Richard C.

    2007-12-01

    Antibody microarrays are an emerging technology that promises to be a powerful tool for the detection of disease biomarkers. The current technology for protein microarrays has been primarily derived from DNA microarrays and is not fully characterized for use with proteins. For example, there are a myriad of surface chemistries that are commercially available for antibody microarrays, but no rigorous studies that compare these different surfaces. Therefore, we have used an enzyme-linked immunosorbent assay (ELISA) microarray platform to analyze 16 different commercially available slide types. Full standard curves were generated for 24 different assays. We found that this approach provides a rigorous and quantitative system for comparing the different slide types based on spot size and morphology, slide noise, spot background, lower limit of detection, and reproducibility. These studies demonstrate that the properties of the slide surface affect the activity of immobilized antibodies and the quality of data produced. Although many slide types can produce useful data, glass slides coated with poly-L-lysine or aminosilane, with or without activation with a crosslinker, consistently produce superior results in the ELISA microarray analyses we performed.

  17. Monoclonal antibodies based on hybridoma technology.

    PubMed

    Yagami, Hisanori; Kato, Hiroshi; Tsumoto, Kanta; Tomita, Masahiro

    2013-03-01

    Based on the size and scope of the present global market for medicine, monoclonal antibodies (mAbs) have a very promising future, with applications for cancers through autoimmune ailments to infectious disease. Since mAbs recognize only their target antigens and not other unrelated proteins, pinpoint medical treatment is possible. Global demand is dramatically expanding. Hybridoma technology, which allows production of mAbs directed against antigens of interest is therefore privileged. However, there are some pivotal points for further development to generate therapeutic antibodies. One is selective generation of human mAbs. Employment of transgenic mice producing human antibodies would overcome this problem. Another focus is recognition sites and conformational epitopes in antigens may be just as important as linear epitopes, especially when membrane proteins such as receptors are targeted. Recognition of intact structures is of critical importance for medical purposes. In this review, we describe patent related information for therapeutic mAbs based on hybridoma technology and also discuss new advances in hybridoma technology that facilitate selective production of stereospecific mAbs. PMID:24237029

  18. Complement in Monoclonal Antibody Therapy of Cancer

    PubMed Central

    Rogers, Laura M.; Veeramani, Suresh; Weiner, George J.

    2015-01-01

    Monoclonal antibodies (mAb) have been used as targeted treatments against cancer for more than a decade, with mixed results. Research is needed to understand mAb mechanisms of action with the goal of improving the efficacy of currently used mAbs, and guiding the design of novel mAbs. While some mAb-induced tumor cell killing is a result of direct effects on tumor cell signaling, mAb opsonization of tumor cells also triggers activation of immune responses due to complement activation and engagement of antibody receptors on immune effector cells. In fact, complement has been shown to play an important role in modulating the anti-tumor activity of many mAb through complement-dependent cytotoxicity (CDC), antibody-dependent cytotoxicity (ADCC), and through indirect effects by modulating the tumor microenvironment. Complement activity can have both agonistic and antagonistic effects on these processes, and which mechanisms are most responsible for effective elimination of malignant cells remain unclear. In this review, we discuss the mAbs currently approved for cancer treatment, and examine how complement can impact their efficacy with a focus on how this information might be used to improve the clinical efficacy of mAb treatment. PMID:24906530

  19. Convergent antibody signatures in human dengue

    PubMed Central

    Parameswaran, Poornima; Liu, Yi; Roskin, Krishna M; Jackson, Katherine KL; Dixit, Vaishali P; Lee, Ji-Yeun; Artiles, Karen; Zompi, Simona; Vargas, Maria José; Simen, Birgitte B; Hanczaruk, Bozena; McGowan, Kim R; Tariq, Muhammad A; Pourmand, Nader; Koller, Daphne; Balmaseda, Angel; Boyd, Scott D; Harris, Eva; Fire, Andrew Z

    2014-01-01

    SUMMARY Dengue is the most prevalent mosquito-transmitted viral disease in humans, and the lack of early prognostics, vaccines and therapeutics contributes to immense disease burden. To identify patterns that could be used for sequence-based monitoring of the antibody response to dengue, we examined antibody heavy-chain gene rearrangements in longitudinal peripheral blood samples from 60 dengue patients. Comparing signatures between acute dengue, post-recovery and healthy samples, we find increased expansion of B cell clones in acute dengue patients, with higher overall clonality in secondary infection. Additionally, we observe consistent antibody sequence features in acute dengue in the major antigen-binding determinant Complementarity Determining Region-3 (CDR3), with specific CDR3 sequences highly enriched in acute samples compared to post-recovery, healthy or non-dengue samples. Dengue thus provides a striking example of a human viral infection where convergent immune signatures can be identified in multiple individuals. Such signatures could facilitate surveillance of immunological memory in communities. PMID:23768493

  20. Antibody Discovery via Multiplexed Single Cell Characterization

    PubMed Central

    Harriman, William D.; Collarini, Ellen J.; Sperinde, Gizette V.; Strandh, Magnus; Fatholahi, Marjan M.; Dutta, April; Lee, Yunji; Mettler, Shelley E.; Keyt, Bruce A.; Ellsworth, Stote L.; Kauvar, Lawrence M.

    2009-01-01

    The secreted immunoglobulin footprint of single hybridoma cells, containing ~10 fg of antibody purified in situ, has been probed for 9 properties concurrently by use of detection labels comprising 280 nm combinatorially colored fluorescent latex beads functionalized with proteins. Specificity of each individual hybridoma cell’s product has thereby been assessed in a primary screen. Varying the density of antigen on beads to modulate the avidity of the interaction between bead and secreted antibody footprint allowed rank ordering by affinity in the same primary screen. As more criteria were added to the selection process, the frequency of positive cells went down; in some cases, the favorable cell was present at <1/50,000. Recovery of the cell of interest was accomplished by plating the cells in a viscous medium on top of a membrane. After collecting the antibody footprint on a capture surface beneath the membrane, the immobilized cells were transferred to an incubator while the footprints were analyzed to locate the hybridoma cells of interest. The desired cells were then cloned by picking them from the corresponding locations on the membrane. PMID:19087879

  1. Demonstration of an antibody-mediated tolerance state and its effect on antibody affinity

    PubMed Central

    1975-01-01

    We have described a model of immunological tolerance induced, in adult mice, by a single injection of a moderate dose of a hapten-protein conjugate. The data suggest that the mechanism of this tolerance state is the production of small amounts of high affinity antibody in response to the tolerance-inducing antigen injection. This antibody acts to inhibit the response to a subsequent challenge with antigen in complete Freund's adjuvant by a mechanism comparable to that of passive antibody-medicated immune suppression. It was shown that a small but high affinity. Tolerance was not terminated by transfer of normal syngeneic spleen or peritoneal cells into tolerant animals. Spleen cells from tolerant mice, when transferred into lethally irradiated, syngeneic animals, produced a PFC response which is greater in magnitude and tolerance state had a significant degree of carrier specificity which was shown to be comparable to the carrier specificity of antibody-mediated immune suppression. hus, evidence was presented to show that one mechanism of tolerance in adult animals in the suppressive effect of small amounts of high affinity antibody formed in response to the tolerizing injection of antigen. PMID:1089745

  2. 60 FR 28616 - Drug Export; Antibody to Hepatitis B Surface Antigen (Human) OrthoSUPTM Antibody to HBsAg Elisa...

    Federal Register 2010, 2011, 2012, 2013, 2014

    1995-06-01

    ...95N-0137] Drug Export; Antibody to Hepatitis B Surface Antigen (Human) Ortho TM...human biological product Antibody to Hepatitis B Surface Antigen (human) ORTHO TM...human biological product Antibody to Hepatitis B Surface Antigen (human) ORTHO...

  3. [Genesis, development and application prospect of antibody library: a review].

    PubMed

    Dai, Heping

    2011-05-01

    Antibodies are immunoglobulins specifically introduced by immunity response of high animals, with the responsibility for recognising and cleaning out specific antigens. Antibody is not only a powerful weapon against pathogen invasion in the organism, but also a tool for specific molecular recognition used in basic scientific research. The diversity of antibody molecules resulted in the concept of antibody library; each individual animal is a natural antibody library. In the post-genome era, in order to fit various "omics", especially for proteomics requirement of high throughput technology, some gene engineering antibody libraries and antibody alternative libraries have been constructed based on phage display technology. Yet, more and more in vitro display systems such as ribosome display, mRNA display have been used for antibody library study, and that present more advantages than phage display. This mini review outlines the genesis, development and application prospect of antibody libraries according to the published reviews and research articles, and offers up to date development and application prospect of antibody library technology. PMID:21845834

  4. Antibody response to myoglobins: effect of host species.

    PubMed

    Cooper, H M; East, I J; Todd, P E; Leach, S J

    1984-06-01

    Using both direct and competitive binding studies it is demonstrated that antibodies to beef myoglobin raised in sheep are able to distinguish between beef and sheep myoglobins although these two proteins differ by only six of the 153 amino acid residues. By contrast, antibodies to beef myoglobin raised in rabbits, dogs and chickens bind almost equally well to beef and sheep myoglobins. It is also shown that antibodies to beef myoglobin raised in sheep have a lower avidity for beef myoglobin than do antibodies raised in more distantly related species. Furthermore, only 50% of the specific anti-beef myoglobin antibodies isolated from sheep antisera will bind to sheep myoglobins whereas 100% of the specific antibodies isolated from the antisera of the other immunised species will bind to sheep myoglobin. It is suggested that antibodies to beef myoglobin are raised to those surface regions which are topographically altered as a result of sequence differences from the host's own myoglobin. When the host animal is evolutionarily distant these sequence differences are considerable and antibodies are raised to the entire surface of the molecule. However, when the host's myoglobin is very similar in sequence to beef myoglobin (as is the case when using sheep as the host animal) antibodies are made only to surface regions affected by the sequence differences. Some of these antibodies--those to the regions of greatest difference--will bind weakly if at all to sheep myoglobin, while those directed to areas of lesser difference will bind well to sheep myoglobin. PMID:6749133

  5. Peptide antibodies and their use in detecting oncogene products

    SciTech Connect

    Wong, G.L.; Arnheim, N.; McCormick, F.P.; Wong, G.L.; Clark, R.; Arnheim, N.; Nitecki, D.E.

    1989-01-17

    A polyclonal antibody preparation is described. It binds selectively to a characteristic marker epitope encompassing amino acid position 12 of an activated form of p21 protein, wherein the polyclonal antibody preparation is specific for a particular polyclonal antibody amino acid at position 12 and does not bind the p21 protein encoded by the corresponding proto-oncogene. A method for detecting an activated form of p21 protein is also described. It is encoded by an oncogene in a cellular sample of a patient which protein has a characteristic marker epitope encompassing amino acid position 12 which is not present in the p21 protein encoded by the corresponding proto-oncogene and which is not exposed in the undenatured protein. The method comprises: (a) treating the sample with a protein denaturing agent that causes the epitope to be exposed and does not substantially inhibit binding of an antibody to the epitope of the protein, (b) incubating the sample with the antibody under conditions that permit the binding of the antibody preparation to the epitope, (c) incubating the sample with a labeled antibody which binds specifically to the antibody employed in step (b), (d) washing the incubated sample to remove unbound labeled antibody, and (e) detecting the presence of labelled immune complexes of the epitope with the antibodies employed in steps (b) and (c).

  6. Profiling antibodies to class II HLA in transplant patient sera.

    PubMed

    McMurtrey, Curtis; Lowe, Dave; Buchli, Rico; Daga, Sunil; Royer, Derek; Humphrey, Alisha; Cate, Steven; Osborn, Sean; Mojsilovic, Aleksandar; VanGundy, Rodney; Bardet, Wilfried; Duty, Andrew; Mojsilovic, Danijela; Jackson, Kenneth; Stastny, Peter; Briggs, David; Zehnder, Daniel; Higgins, Rob; Hildebrand, William

    2014-03-01

    Immunizing events including pregnancy, transfusions, and transplantation promote strong alloantibody responses to HLA. Such alloantibodies to HLA preclude organ transplantation, foster hyperacute rejection, and contribute to chronic transplant failure. Diagnostic antibody-screening assays detect alloreactive antibodies, yet key attributes including antibody concentration and isotype remain largely unexplored. The goal here was to provide a detailed profile of allogeneic antibodies to class II HLA. Methodologically, alloantibodies were purified from sensitized patient sera using an HLA-DR11 immunoaffinity column and subsequently categorized. Antibodies to DR11 were found to fix complement, exist at a median serum concentration of 2.3?g/mL, consist of all isotypes, and isotypes IgG2, IgM, and IgE were elevated. Because multimeric isotypes can confound diagnostic determinations of antibody concentration, IgM and IgA isotypes were removed and DR11-IgG tested alone. Despite removal of multimeric isotypes, patient-to-patient antibody concentrations did not correlate with MFI values. In conclusion, allogeneic antibody responses to DR11 are comprised of all antibody isotypes at differing proportions, these combined isotypes fix complement at nominal serum concentrations, and enhancements other than the removal of IgM and IgA multimeric isotypes may be required if MFI is to be used as a means of determining anti-HLA serum antibody concentrations in diagnostic clinical assays. PMID:24269696

  7. Anti-idiotypic antibodies induce neutralizing antibodies to bovine herpesvirus 1.

    PubMed Central

    Srikumaran, S; Onisk, D V; Borca, M V; Nataraj, C; Zamb, T J

    1990-01-01

    A neutralizing murine monoclonal antibody (mAb) of the IgG2a isotype (MM-113), specific for bovine herpesvirus 1 (BHV-1) glycoprotein gIV, was used to develop anti-idiotypic antibodies (anti-Id) in a calf. The bovine anti-Id were isolated from the serum of the immunized calf by affinity chromatography on an MM-113-Sepharose column, followed by repeated adsorption on a murine IgG2a column. The anti-Id thus obtained specifically reacted with MM-113, but not with isotype-matched controls. They also inhibited the binding of MM-113 to BHV-1 in a concentration-dependent manner. Mice immunized with the anti-Id produced neutralizing antibodies to BHV-1. The anti-Id bound to cells permissive to BHV-1 in a cell-binding radioimmunoassay (RIA). PMID:2165998

  8. Generation of neutralising antibodies against porcine endogenous retroviruses (PERVs)

    SciTech Connect

    Kaulitz, Danny; Fiebig, Uwe; Eschricht, Magdalena; Wurzbacher, Christian; Kurth, Reinhard; Denner, Joachim, E-mail: DennerJ@rki.d

    2011-03-01

    Antibodies neutralising porcine endogenous retroviruses (PERVs) were induced in different animal species by immunisation with the transmembrane envelope protein p15E. These antibodies recognised epitopes, designated E1, in the fusion peptide proximal region (FPPR) of p15E, and E2 in the membrane proximal external region (MPER). E2 is localised in a position similar to that of an epitope in the transmembrane envelope protein gp41 of the human immunodeficiency virus-1 (HIV-1), recognised by the monoclonal antibody 4E10 that is broadly neutralising. To detect neutralising antibodies specific for PERV, a novel assay was developed, which is based on quantification of provirus integration by real-time PCR. In addition, for the first time, highly effective neutralising antibodies were obtained by immunisation with the surface envelope protein of PERV. These data indicate that neutralising antibodies can be induced by immunisation with both envelope proteins.

  9. Antigen-Antibody Testing: A Visual Simulation or Virtual Reality

    NSDL National Science Digital Library

    Daniel Schadler

    2011-01-01

    In this biology activity, learners use plastic pipettes to cut wells into the solid gel layer of agar in petri dishes and place solutions of simulated antigen and antibody preparations into the wells. The antigens and antibodies diffuse into the gel layer and react to form a precipitate. This activity demonstrates the biological phenomenon of the formation of a precipitate when an antigen reacts with an antibody. The exercise can be used to illustrate the specificity of antigen-antibody reactions, showing that a precipitation reaction only occurs when an antibody reacts with the antigen that was used to induce the formation of the antibody. The exercise is also a general demonstration of diffusion. Adult supervision is recommended.

  10. Cerebrospinal Fluid Aquaporin-4 Antibody Levels in Neuromyelitis Optica Attacks

    PubMed Central

    Sato, Douglas Kazutoshi; Callegaro, Dagoberto; de Haidar Jorge, Frederico M; Nakashima, Ichiro; Nishiyama, Shuhei; Takahashi, Toshiyuki; Simm, Renata Faria; Apostolos-Pereira, Samira Luisa; Misu, Tatsuro; Steinman, Lawrence; Aoki, Masashi; Fujihara, Kazuo

    2014-01-01

    To elucidate immunopathogenetic roles of aquaporin-4 antibodies in the cerebrospinal fluid (CSF) of neuromyelitis optica spectrum disorders (NMOSD), we analyzed aquaporin-4 antibody titers, cellular and inflammatory markers in the CSF collected from 11 aquaporin-4 antibody seropositive patients. The CSF aquaporin-4 antibody levels during attacks (but not in sera) closely correlated with pleocytosis, inflammatory cytokines including interleukin-6 that can regulate antibody-producing plasmablasts, and glial fibrillary acidic protein levels in the CSF. The amount of aquaporin-4 antibodies present in the central nervous system may have therapeutic implications, as it is associated with astrocyte injury and inflammatory responses during NMOSD attacks. Ann Neurol 2014;76:305–309 PMID:24977390

  11. Atypical Miller Fisher syndrome with GQ1b antibodies

    Microsoft Academic Search

    Mark A Paine; Geoff Keir; Gordon T Plant

    1996-01-01

    An atypical case of Miller Fisher syndrome is described in a patient with ophthalmoplegia and mild ataxia but no areflexia. High titres of acute phase antibodies to gangliosides asialo-GM1 and GQ1b were detected. Asialo-GM1 antibodies have not been previously reported in association with Miller Fisher syndrome. Considerable clinical recovery occurred in association with reduction in the ganglioside antibody titres. Ganglioside

  12. Serial plasmapheresis in a haemophiliac with antibodies to FVIII

    Microsoft Academic Search

    R Cobcroft; G Tamagnini; K M Dormandy

    1977-01-01

    Serial plasmaphereses were performed on a 23-year-old haemophiliac, with antibodies to factor VIII (FVIII), in order to reduce the antibody level before multiple dental extractions. Eight phereses were carried out in which approximately 1.5 litres plasma was exchanged with isotonic saline. By the ninth exchange, which was carried out immediately before the extraction, the antibody level had fallen from 4

  13. An update on the use of antibodies against the filoviruses

    PubMed Central

    Saphire, Erica Ollmann

    2015-01-01

    Multiple recent, independent studies have confirmed that passively administered antibodies can provide effective postexposure therapy in nonhuman primates after exposure to an otherwise lethal dose of Ebola virus or Marburg virus. In this article, we review composition and performance of the antibody cocktails tested thus far, what is known about antibody epitopes on the viral glycoprotein target and ongoing research questions in further development of such cocktails for pre-exposure or emergency postexposure use. PMID:24188676

  14. Identification of Pigment Cell Antigens Defined by Vitiligo Antibodies

    Microsoft Academic Search

    Jian Cui; Ronald Harning; Milagros Henn; Jean-Claude Bystryn

    1992-01-01

    Patients with vitiligo have circulating antibodies to pigment cells. To characterize this response further and to identify the antigens defined by vitiligo antibodies, sera of 23 patients with vitiligo and 22 patients with unrelated conditions were analyzed by immunoprecipitation and SDS-PAGE analysis of 125I-labeled cell antigens on pigment and control cells. Antibodies to pigment cell antigens were present in 18

  15. Effect of anti-lymphocyte serum on natural antibody

    PubMed Central

    Muschel, L. H.; Gustafson, Linda; Atai, M.

    1968-01-01

    Horse anti-dog lymphocyte serum was found to suppress the immune response of dogs normally resulting from the injection of Salmonella typhi vaccine. Normal antibody levels of the dogs against Shigella dysenteriae were unaffected, however, by the anti-lymphocyte serum. The results suggest, therefore, that the natural antibodies may arise by a population of cells or by mechanisms different from those which produce antibodies as a result of deliberate antigenic stimulation. PMID:4867939

  16. Renaissance of the Blocking Antibody Concept in Type I Allergy

    Microsoft Academic Search

    Sabine Flicker; Rudolf Valenta

    2003-01-01

    Formation of IgE antibodies against per se harmless antigens (i.e. allergens) is the hallmark and key pathomechanism of type I allergy, a hypersensitivity disease affecting more than 25% of the population. Classical experiments performed more than 65 years ago demonstrated that allergen-specific IgG antibodies, termed blocking antibodies, can antagonize the cascade of allergic inflammation resulting from allergen recognition by IgE

  17. Detection of serum antibodies against Chlamydia pneumoniae by ELISA

    Microsoft Academic Search

    Kei Numazaki; Tadashi Ikebe; Shunzo Chiba

    1996-01-01

    Chlamydia pneumoniae causes pneumonia and other respiratory infections in children, adolescents and adults. We tried to evaluate the diagnostic value of detection of serum antibodies by ELISA for C. pneumoniae infections in Japanese children. Serum IgG, IgA and IgM antibodies to C. pneumoniae were determined by the microimmunofluorescence (MIF) test. Serum IgG and IgA antibodies were also determined by ELISA

  18. High-throughput generation and engineering of recombinant human antibodies

    Microsoft Academic Search

    Barbara Krebs; Robert Rauchenberger; Silke Reiffert; Christine Rothe; Michael Tesar; Elisabeth Thomassen; Manqiu Cao; Torsten Dreier; David Fischer; Adolf Höß; Landon Inge; Achim Knappik; Matthias Marget; Peter Pack; Xian-Qin Meng; Robert Schier; Peter Söhlemann; Jill Winter; Joachim Wölle; Titus Kretzschmar

    2001-01-01

    The first version of the Human Combinatorial Antibody Library (HuCAL®) is a single-chain Fv-based phage display library (HuCAL®-scFv) with 2×109 members optimised for high-throughput generation and targeted engineering of human antibodies. 61% of the library genes code for functional scFv as judged by sequencing. We show here that since HuCAL®-scFv antibodies are expressed in high levels in Escherichia coli, automated

  19. Second Generation Anti-MUCl Peptide Monoclonal Antibodies

    Microsoft Academic Search

    Pei-xiang Xing; Julie Prenzoska; Kaylene Quelch; Ian F. C. McKenzie

    Second generation antibodies to mammary mucins were produced by immunizing mice with a peptide with a sequence deduced from that of the MUC1 complementary DNA sequence (PAHGVTSAPDTRPAPGS- TAP). Four monoclonal antibodies (BCP7-10) were produced which gave different reactions. BCP8 was similar in tissue reactivity (by immunoper- oxidase staining) to anti-breast cancer or anti-human milk fat globule membranes (HMFG) antibodies and

  20. Monoclonal antibody therapy for prostate cancer.

    PubMed

    Jakobovits, A

    2008-01-01

    Early detection of prostate cancer (PCa) and advances in hormonal and chemotherapy treatments have provided great clinical benefits to patients with early stages of the disease. However, a significant proportion of patients still progress to advanced, metastatic disease, for which no effective therapies are available. Therefore, there is a critical need for new treatment modalities, ideally targeted specifically to prostate cancer cells. The recent clinical and commercial successes of monoclonal antibodies (MAbs) have made them the most rapidly expanding class of therapeutics being developed for many disease indications, including cancer. PCa is well suited for antibody-based therapy due to the size and location of recurrent and metastatic tumors, and the lack of necessity to avoid targeting the normal prostate, a nonessential organ. These properties have fostered interest in the development and clinical evaluation of therapeutic MAbs directed to both well established and newly discovered targets in PCa. MAbs directed to established targets include those approved for other solid tumors, including anti-human epidermal growth factor receptor-2 (HER2) MAb trastuzumab, anti-epidermal growth factor receptor (EGFR) MAbs cetuximab and panitumumab, and the antivascular endothelial growth factor (VEGF) MAb bevacizumab. Genomics efforts have yielded a large number of novel, clinically relevant targets in PCa with the desirable expression profiling in tumor and normal tissues, and with an implicated role in tumor growth and spread. Growing efforts are directed to the development of naked or payload-conjugated therapeutic antibodies to these targets, and a variety of MAb products are currently progressing through preclinical and various stages of clinical development. The clinical experience with some of the commercialized MAb products points out specific challenges in conducting clinical trials with targeted therapy in PCa. PMID:18071949

  1. Derivatized gold clusters and antibody-gold cluster conjugates

    DOEpatents

    Hainfeld, James F. (Shoreham, NY); Furuya, Frederic R. (Williston Park, NY)

    1994-11-01

    Antibody- or antibody fragment-gold cluster conjugates are shown wherein the conjugate size can be as small as 5.0 nm. Methods and reagents are disclosed in which antibodies, Fab' or F(ab').sub.2 fragments thereof are covalently bound to a stable cluster of gold atoms. The gold clusters may contain 6, 8, 9, 11, 13, 55 or 67 gold atoms in their inner core. The clusters may also contain radioactive gold. The antibody-cluster conjugates are useful in electron microscopy applications as well as in clinical applications that include imaging, diagnosis and therapy.

  2. 2nd PEGS Annual Symposium on Antibodies for Cancer Therapy

    PubMed Central

    Ho, Mitchell; Royston, Ivor; Beck, Alain

    2012-01-01

    The 2nd Annual Antibodies for Cancer Therapy symposium, organized again by Cambridge Healthtech Institute as part of the Protein Engineering Summit, was held in Boston, USA from April 30th to May 1st, 2012. Since the approval of the first cancer antibody therapeutic, rituximab, fifteen years ago, eleven have been approved for cancer therapy, although one, gemtuzumab ozogamicin, was withdrawn from the market.  The first day of the symposium started with a historical review of early work for lymphomas and leukemias and the evolution from murine to human antibodies. The symposium discussed the current status and future perspectives of therapeutic antibodies in the biology of immunoglobulin, emerging research on biosimilars and biobetters, and engineering bispecific antibodies and antibody-drug conjugates. The tumor penetration session was focused on the understanding of antibody therapy using ex vivo tumor spheroids and the development of novel agents targeting epithelial junctions in solid tumors. The second day of the symposium discussed the development of new generation recombinant immunotoxins with low immunogenicity, construction of chimeric antigen receptors, and the proof-of-concept of ‘photoimmunotherapy’. The preclinical and clinical session presented antibodies targeting Notch signaling and chemokine receptors.  Finally, the symposium discussed emerging technologies and platforms for therapeutic antibody discovery. PMID:22864478

  3. Mechanisms of monoclonal antibody stabilization and release from silk biomaterials

    PubMed Central

    Guziewicz, Nicholas A.; Massetti, Andrew J.; Perez-Ramirez, Bernardo J.; Kaplan, David L.

    2013-01-01

    The availability of stabilization and sustained delivery systems for antibody therapeutics remains a major clinical challenge, despite the growing development of antibodies for a wide range of therapeutic applications due to their specificity and efficacy. A mechanistic understanding of protein-matrix interactions is critical for the development of such systems and is currently lacking as a mode to guide the field. We report mechanistic insight to address this need by using well-defined matrices based on silk gels, in combination with a monoclonal antibody. Variables including antibody loading, matrix density, charge interactions, hydrophobicity and water access were assessed to clarify mechanisms involved in the release of antibody from the biomaterial matrix. The results indicate that antibody release is primarily governed by hydrophobic interactions and hydration resistance, which are controlled by silk matrix chemistry, peptide domain distribution and protein density. Secondary ionic repulsions are also critical in antibody stabilization and release. Matrix modification by free methionine incorporation was found to be an effective strategy for mitigating encapsulation induced antibody oxidation. Additionally, these studies highlight a characterization approach to improve the understanding and development of other protein sustained delivery systems, with broad applicability to the rapidly developing monoclonal antibody field. PMID:23859659

  4. Method for detecting pathogens attached to specific antibodies

    DOEpatents

    Miles, Robin R.; Venkateswaran, Kodumudi S.; Fuller, Christopher K.

    2005-01-25

    The use of impedance measurements to detect the presence of pathogens attached to antibody-coated beads. In a fluidic device antibodies are immobilized on a surface of a patterned interdigitated electrode. Pathogens in a sample fluid streaming past the electrode attach to the immobilized antibodies, which produces a change in impedance between two adjacent electrodes, which impedance change is measured and used to detect the presence of a pathogen. To amplify the signal, beads coated with antibodies are introduced and the beads would stick to the pathogen causing a greater change in impedance between the two adjacent electrodes.

  5. Modern Technologies for Creating Synthetic Antibodies for Clinical application

    PubMed Central

    Lebedenko, E. N.

    2009-01-01

    The modular structure and versatility of antibodies enables one to modify natural immunoglobulins in different ways for various clinical applications. Rational design and molecular engineering make it possible to directionally modify the molecular size, affinity, specificity, and immunogenicity and effector functions of an antibody, as well as to combine them with other functional agents. This review focuses on up-to-date methods of antibody engineering for diagnosing and treating various diseases, particularly on new technologies meant to refine the effector functions of therapeutic antibodies. PMID:22649585

  6. Nature-inspired design of motif-specific antibody scaffolds

    PubMed Central

    Koerber, James T.; Thomsen, Nathan D.; Hannigan, Brett T.; Degrado, William F.; Wells, James A.

    2013-01-01

    Aberrant changes in post-translational modifications (PTMs) such as phosphorylation underlie a majority of human diseases. However, detection and quantification of PTMs for diagnostic or biomarker applications often requires monoclonal PTM-specific antibodies, which are challenging to generate using traditional antibody-generation platforms. Here we outline a general strategy for producing synthetic PTM-specific antibodies by engineering a motif-specific ‘hot spot’ into an antibody scaffold. Inspired by a natural phosphate-binding motif, we designed antibody scaffolds with hot spots specific for phosphoserine, phosphothreonine, or phosphotyrosine. Crystal structures of the phospho-specific antibodies revealed two distinct modes of phosphoresidue recognition. Our data suggest that each hot spot functions independently of the surrounding scaffold, as phage display antibody libraries using these scaffolds yielded >50 phospho- and target-specific antibodies against 70% of target peptides. Ultimately, our motif-specific scaffold strategy may provide a general solution for the rapid, robust development of monoclonal anti-PTM antibodies for signaling, diagnostic and therapeutic applications. PMID:23955275

  7. Impedance measurements for detecting pathogens attached to antibodies

    DOEpatents

    Miles, Robin R.; Venkateswaran, Kodumudi S.; Fuller, Christopher K.

    2004-12-28

    The use of impedance measurements to detect the presence of pathogens attached to antibody-coated beads. In a fluidic device antibodies are immobilized on a surface of a patterned interdigitated electrode. Pathogens in a sample fluid streaming past the electrode attach to the immobilized antibodies, which produces a change in impedance between two adjacent electrodes, which impedance change is measured and used to detect the presence of a pathogen. To amplify the signal, beads coated with antibodies are introduced and the beads would stick to the pathogen causing a greater change in impedance between the two adjacent electrodes.

  8. Sequencing the functional antibody repertoire—diagnostic and therapeutic discovery

    PubMed Central

    Robinson, William H.

    2015-01-01

    The development of high-throughput DNA sequencing technologies has enabled large-scale characterization of functional antibody repertoires, a new method of understanding protective and pathogenic immune responses. Important parameters to consider when sequencing antibody repertoires include the methodology, the B-cell population and clinical characteristics of the individuals analysed, and the bioinformatic analysis. Although focused sequencing of immunoglobulin heavy chains or complement determining regions can be utilized to monitor particular immune responses and B-cell malignancies, high-fidelity analysis of the full-length paired heavy and light chains expressed by individual B cells is critical for characterizing functional antibody repertoires. Bioinformatic identification of clonal antibody families and recombinant expression of representative members produces recombinant antibodies that can be used to identify the antigen targets of functional immune responses and to investigate the mechanisms of their protective or pathogenic functions. Integrated analysis of coexpressed functional genes provides the potential to further pinpoint the most important antibodies and clonal families generated during an immune response. Sequencing antibody repertoires is transforming our understanding of immune responses to autoimmunity, vaccination, infection and cancer. We anticipate that antibody repertoire sequencing will provide next-generation biomarkers, diagnostic tools and therapeutic antibodies for a spectrum of diseases, including rheumatic diseases. PMID:25536486

  9. Salivary IgA antibody to glucosyltransferase in man.

    PubMed

    Smith, D J; Taubman, M A; Ebersole, J L

    1985-08-01

    Parotid salivas of 97 young adults were screened for IgA antibody to glucosyltransferase (GTF) from laboratory strains of Streptococcus mutans (serotypes c and g). Antibody levels to GTF from serotype c positively correlated with levels to serotype g GTF among these salivas. GTF's were prepared from S. mutans obtained from a subset of individuals in this population. All but one saliva showed IgA antibody activity to all of the GTF tested. In addition, the relative magnitude of each subject's antibody level was generally the highest to the GTF from their own S. mutans. Fractions, enriched for IgA by ammonium sulphate precipitation and gel filtration, showed patterns of functional inhibition of GTF activity which were consistent with patterns of IgA antibody activity in ELISA of unfractionated salivas. These data indicate that detectable levels of IgA antibody to S. mutans GTF exist in many young adult salivas, while this IgA antibody activity reacts with GTF from different biotypes, subjects generally show the highest secretory IgA antibody levels to their own GTF, and the relative amount of IgA antibody to GTF and the ability to inhibit GTF activity are roughly correlated. PMID:2931224

  10. Salivary IgA antibody to glucosyltransferase in man.

    PubMed Central

    Smith, D J; Taubman, M A; Ebersole, J L

    1985-01-01

    Parotid salivas of 97 young adults were screened for IgA antibody to glucosyltransferase (GTF) from laboratory strains of Streptococcus mutans (serotypes c and g). Antibody levels to GTF from serotype c positively correlated with levels to serotype g GTF among these salivas. GTF's were prepared from S. mutans obtained from a subset of individuals in this population. All but one saliva showed IgA antibody activity to all of the GTF tested. In addition, the relative magnitude of each subject's antibody level was generally the highest to the GTF from their own S. mutans. Fractions, enriched for IgA by ammonium sulphate precipitation and gel filtration, showed patterns of functional inhibition of GTF activity which were consistent with patterns of IgA antibody activity in ELISA of unfractionated salivas. These data indicate that detectable levels of IgA antibody to S. mutans GTF exist in many young adult salivas, while this IgA antibody activity reacts with GTF from different biotypes, subjects generally show the highest secretory IgA antibody levels to their own GTF, and the relative amount of IgA antibody to GTF and the ability to inhibit GTF activity are roughly correlated. PMID:2931224

  11. Commercially available angiotensin II At? receptor antibodies are nonspecific.

    PubMed

    Hafko, Roman; Villapol, Sonia; Nostramo, Regina; Symes, Aviva; Sabban, Esther L; Inagami, Tadashi; Saavedra, Juan M

    2013-01-01

    Commercially available angiotensin II At? receptor antibodies are widely employed for receptor localization and quantification, but they have not been adequately validated. In this study, we characterized three commercially available At? receptor antibodies: 2818-1 from Epitomics, sc-9040 from Santa Cruz Biotechnology, Inc., and AAR-012 from Alomone Labs. Using western blot analysis the immunostaining patterns observed were different for every antibody tested, and in most cases consisted of multiple immunoreactive bands. Identical immunoreactive patterns were present in wild-type and At? receptor knockout mice not expressing the target protein. In the mouse brain, immunocytochemical studies revealed very different cellular immunoreactivity for each antibody tested. While the 2818-1 antibody reacted only with endothelial cells in small parenchymal arteries, the sc-9040 antibody reacted only with ependymal cells lining the cerebral ventricles, and the AAR-012 antibody reacted only with multiple neuronal cell bodies in the cerebral cortex. Moreover, the immunoreactivities were identical in brain tissue from wild-type or At? receptor knockout mice. Furthermore, in both mice and rat tissue extracts, there was no correlation between the observed immunoreactivity and the presence or absence of At? receptor binding or gene expression. We conclude that none of these commercially available At? receptor antibodies tested met the criteria for specificity. In the absence of full antibody characterization, competitive radioligand binding and determination of mRNA expression remain the only reliable approaches to study At? receptor expression. PMID:23840911

  12. [The Ro/SS-A antigen-antibody system].

    PubMed

    Mayet, W J; Bachmann, M; Hermann, E; Poralla, T; Müller, W E; Meyer zum Büschenfelde, K H

    1988-01-01

    First described in 1969 by Clark et al., the Ro/SS-A antibody system has proved most important in the evaluation of lupus patients possessing prominent photosensitive cutaneous lesions. Ro antibodies are also seen in patients with primary Sjögren's syndrome, neonatal lupus erythematosus, congenital heart block and ANA-negative LE. The most recent data of biochemical characterization have led to the supposition that these antibodies may exert a direct pathogenic effect. In this review the current knowledge concerning the Ro-antigen-antibody system is summarized. PMID:3291481

  13. The Role of Monoclonal Antibodies in the Management of Leukemia

    PubMed Central

    Al-Ameri, Ali; Cherry, Mohamad; Al-Kali, Aref; Ferrajoli, Alessandra

    2010-01-01

    This article will review the monoclonal antibodies more commonly used in leukemias. In the last three decades, scientists have made considerable progress understanding the structure and the functions of various surface antigens, such as CD20, CD33. The introduction of rituximab, an anti CD20 monoclonal antibody, had a great impact in the treatment of lymphoproliferative disorders. Gemtuzumab, an anti CD 33 conjugated monoclonal antibody has activity in acute mylegenous leukemia (AML). As this field is undergoing a rapid growth, the years will see an increasing use of monoclonal antibodies in hematological malignancies.

  14. Monoclonal antibodies in the treatment of immune thrombocytopenic purpura (ITP).

    PubMed

    Gómez-Almaguer, David

    2012-04-01

    Immune thrombocytopenic purpura is characterized by antibody-mediated destruction of platelets and suboptimal platelet production. Initially the treatment of ITP includes corticosteroids, IgG-anti-D, and intravenous immunoglobulins. Splenectomy and monoclonal antibodies are usually considered for refractory and chronic ITP patients. There are new data suggesting that early administration of rituximab is important, and this antibody has been used as first-line therapy in adults. In this concise review the role of rituximab and other monoclonal antibodies is analyzed. These agents have the capability of sparing splenectomy and possibly curing the disease in some patients. PMID:22507772

  15. Chimeric antibodies with extended half-life in ferrets

    PubMed Central

    Nesspor, Thomas C; Scallon, Bernard

    2014-01-01

    Background Ferrets have long been used as a disease model for the study of influenza vaccines, but a more recent use has been for the study of human monoclonal antibodies directed against influenza viruses. Published data suggest that human antibodies are cleared unusually quickly from the ferret and that immune responses may be partially responsible. This immunogenicity increases variability within groups and may present an obstacle to long-term studies. Objective Our aim was to identify an antibody design with reduced immunogenicity and longer circulating half-life in ferrets. Methods The constant region coding sequences for ferret immunoglobulin G were cloned, and chimeric human/ferret antibodies were expressed and purified. Some of the chimeric antibodies included substitutions that have been shown to extend the half-life of human IgG antibodies. These chimeric antibodies were tested for binding to recombinant ferret FcRn receptor and then evaluated in pharmacokinetic studies in ferrets. Results A one-residue substitution in the ferret Fc domain, S252Y, was identified that increased binding affinity to the ferret neonatal receptor by 24-fold and extended half-life from 65 ± 27 to 206 ± 28 hours or ?9 days. Ferrets dosed twice with this surrogate antibody showed no indications of an immune response. Conclusion Expressing the variable region of a candidate human therapeutic antibody with ferret constant regions containing the S252Y substitution can offer long half-life and limit immunogenicity. PMID:25074755

  16. Adeno-Associated Virus Delivery of Broadly Neutralizing Antibodies

    PubMed Central

    Schnepp, Bruce C.; Johnson, Philip R.

    2014-01-01

    Purpose of review In this review, we will discuss the emerging field of vector mediated antibody gene transfer as an alternative HIV vaccine. This approach is an improvement over classical passive immunization strategies that administer antibodies to the host to provide protection from infection. With vector mediated gene transfer, the antibody gene is delivered to the host resulting in long-term endogenous antibody expression from the injected muscle that confers protective immunity. Recent Findings A large number of very potent and broadly neutralizing HIV antibodies have recently been isolated and characterized. Vector mediated antibody gene transfer allows one to immediately use these antibodies as a vaccine. Gene transfer studies in both mice and monkeys demonstrate long-term antibody expression in serum from a single injection at concentrations that provide sterilizing immunity. Summary Vector mediated antibody gene transfer can rapidly move existing, potent anti-HIV molecules into the clinic. The gene transfer products demonstrate a potency and breadth identical to the original product. This strategy eliminates the need for immunogen design and interaction with the adaptive immune system to generate protection, a strategy that so far has shown little promise. PMID:24638019

  17. Quantitation of dengue precipitating antibody by inhibition countercurrent immunoelectrophoresis.

    PubMed Central

    Churdboonchart, V; Bhamarapravati, N; Harisdangkul, V; Futrakul, P; Chiengsong, R

    1981-01-01

    The inhibition countercurrent immunoelectrophoresis test was employed to detect dengue virus antibody in patients' sera. Anti-dengue type 2 titers determined by inhibition countercurrent immunoelectrophoresis correlated well with hemagglutination inhibition titers. In secondary cases, more than fourfold increases in precipitating antibodies were observed. The control sera were negative except for sera from a few patients with systemic lupus erythematosus, which showed low titers. Simultaneous detection of dengue virus antigen and antibody in sera collected during the acute phase could confirm at least 90% of cases. This method is recommended as a routine technique to quantitate antibody in sera from suspected cases of dengue hemorrhagic fever. PMID:7240383

  18. Derivatized gold clusters and antibody-gold cluster conjugates

    DOEpatents

    Hainfeld, J.F.; Furuya, F.R.

    1994-11-01

    Antibody- or antibody fragment-gold cluster conjugates are shown wherein the conjugate size can be as small as 5.0 nm. Methods and reagents are disclosed in which antibodies, Fab' or F(ab')[sub 2] fragments are covalently bound to a stable cluster of gold atoms. The gold clusters may contain 6, 8, 9, 11, 13, 55 or 67 gold atoms in their inner core. The clusters may also contain radioactive gold. The antibody-cluster conjugates are useful in electron microscopy applications as well as in clinical applications that include imaging, diagnosis and therapy. 7 figs.

  19. The challenges of modelling antibody repertoire dynamics in HIV infection.

    PubMed

    Luo, Shishi; Perelson, Alan S

    2015-09-01

    Antibody affinity maturation by somatic hypermutation of B-cell immunoglobulin variable region genes has been studied for decades in various model systems using well-defined antigens. While much is known about the molecular details of the process, our understanding of the selective forces that generate affinity maturation are less well developed, particularly in the case of a co-evolving pathogen such as HIV. Despite this gap in understanding, high-throughput antibody sequence data are increasingly being collected to investigate the evolutionary trajectories of antibody lineages in HIV-infected individuals. Here, we review what is known in controlled experimental systems about the mechanisms underlying antibody selection and compare this to the observed temporal patterns of antibody evolution in HIV infection. We describe how our current understanding of antibody selection mechanisms leaves questions about antibody dynamics in HIV infection unanswered. Without a mechanistic understanding of antibody selection in the context of a co-evolving viral population, modelling and analysis of antibody sequences in HIV-infected individuals will be limited in their interpretation and predictive ability. PMID:26194760

  20. Monoclonal antibodies against chicken interleukin-6

    Microsoft Academic Search

    T. R. Scott; H. S. Lillehoj

    2006-01-01

    Monoclonal antibodies (mAb) were produced against a recombinant (r) chicken interleukin-6 (IL-6). Eight mAbs produced were tested for isotype; ability to inhibit recombinant forms of chicken (ch), human (h) and murine (m) IL-6; and recognition of rchIL-6 by Western immunoblotting. The mAb isotypes were represented by IgG1 (one), IgG2a (six) and IgG2b (one). In a mouse B9 hybridoma cell bioassay

  1. [Natalizumab: an antibody targeting ?4-integrin].

    PubMed

    Nakahara, Jin

    2014-10-01

    Multiple sclerosis (MS) is a demyelinating disease of the central nervous system (CNS), in which inflammation develops upon leukocyte invasion into the CNS via the blood-brain barrier. ?4-Integrin is an important cell adhesion molecule involved in the penetration process. Natalizumab is an ?4-integrin-targeting monoclonal antibody that was recently approved for use as a disease-modifying therapy for MS in Japan. In this article, the mechanism of action, efficacy, and side effects of natalizumab will be reviewed. PMID:25296869

  2. Phylogenetic study of transcortin using monoclonal antibodies.

    PubMed

    Faict, D; De Moor, P

    1986-08-14

    We produced monoclonal antibodies that recognise three distinct epitopes of human transcortin. These epitopes are present on transcortin of humans with normal and altered transcortin levels, as well as on a variant with lower affinity for cortisol. One epitope is present on transcortin of Old World Monkeys and apes, the others are only present on transcortin of apes. The epitopes are not present on transcortin of other species. These results indicate that human transcortin contains a highly evolved and a more conserved part. PMID:2428359

  3. Antibody zymography: a novel adaptation of zymography to determine the protease-neutralising potential of specific antibodies and snake antivenoms.

    PubMed

    Hasson, S S; Theakston, R D G; Harrison, R A

    2004-09-01

    A common problem in the development of antibody-based therapeutics is the selection, usually from a large population, of specific antibodies with the desired function. One of our research objectives is to identify antibodies capable of neutralising the most important haemorrhagic and haemostasis-disruptive proteases from viper venom. Here, we describe a modification of conventional gelatin-zymography that permits the identification of antibodies capable of neutralising gelatinolytic proteases. We demonstrate that the gelatinolytic activity of viper venom proteases is neutralised by addition of viper antivenom to the matrix of conventional gelatin-zymograms. Venom protein gelatinolytic activity was unaffected by inclusion of antibody from control, non-immunised animals or immunoglobulin-depleted serum. The application of this antibody zymogram technique for future research on snake venoms is evaluated in the context of identified limitations. PMID:15350518

  4. An Insertion Mutation That Distorts Antibody Binding Site Architecture Enhances Function of a Human Antibody

    SciTech Connect

    Krause, Jens C.; Ekiert, Damian C.; Tumpey, Terrence M.; Smith, Patricia B.; Wilson, Ian A.; Crowe, Jr., James E. (Vanderbilt); (Scripps); (CDC)

    2011-09-02

    The structural and functional significance of somatic insertions and deletions in antibody chains is unclear. Here, we demonstrate that a naturally occurring three-amino-acid insertion within the influenza virus-specific human monoclonal antibody 2D1 heavy-chain variable region reconfigures the antibody-combining site and contributes to its high potency against the 1918 and 2009 pandemic H1N1 influenza viruses. The insertion arose through a series of events, including a somatic point mutation in a predicted hot-spot motif, introduction of a new hot-spot motif, a molecular duplication due to polymerase slippage, a deletion due to misalignment, and additional somatic point mutations. Atomic resolution structures of the wild-type antibody and a variant in which the insertion was removed revealed that the three-amino-acid insertion near the base of heavy-chain complementarity-determining region (CDR) H2 resulted in a bulge in that loop. This enlarged CDR H2 loop impinges on adjacent regions, causing distortion of the CDR H1 architecture and its displacement away from the antigen-combining site. Removal of the insertion restores the canonical structure of CDR H1 and CDR H2, but binding, neutralization activity, and in vivo activity were reduced markedly because of steric conflict of CDR H1 with the hemagglutinin antigen.

  5. Mathematical and Experimental Analyses of Antibody Transport in Hollow-Fiber-Based Specific Antibody Filters

    E-print Network

    Federspiel, William J.

    specificity directly from whole blood, without separation of the plasma and cellular blood components) that selectively remove antibodies of a given specificity directly from whole blood, without separation of the plasma and cellular blood components and with minimal removal of plasma proteins other than the targeted

  6. Selection and identification of single domain antibody fragments from camel heavy-chain antibodies

    Microsoft Academic Search

    M Arbabi Ghahroudi; A Desmyter; L Wyns; R Hamers; S Muyldermans

    1997-01-01

    Functional heavy-chain ?-immunoglobulins lacking light chains occur naturally in Camelidae. We now show the feasibility of immunising a dromedary, cloning the repertoire of the variable domains of its heavy-chain antibodies and panning, leading to the successful identification of minimum sized antigen binders. The recombinant binders are expressed well in E. coli, extremely stable, highly soluble, and react specifically and with

  7. An Insertion Mutation That Distorts Antibody Binding Site Architecture Enhances Function of a Human Antibody

    PubMed Central

    Krause, Jens C.; Ekiert, Damian C.; Tumpey, Terrence M.; Smith, Patricia B.; Wilson, Ian A.; Crowe, James E.

    2011-01-01

    The structural and functional significance of somatic insertions and deletions in antibody chains is unclear. Here, we demonstrate that a naturally occurring three-amino-acid insertion within the influenza virus-specific human monoclonal antibody 2D1 heavy-chain variable region reconfigures the antibody-combining site and contributes to its high potency against the 1918 and 2009 pandemic H1N1 influenza viruses. The insertion arose through a series of events, including a somatic point mutation in a predicted hot-spot motif, introduction of a new hot-spot motif, a molecular duplication due to polymerase slippage, a deletion due to misalignment, and additional somatic point mutations. Atomic resolution structures of the wild-type antibody and a variant in which the insertion was removed revealed that the three-amino-acid insertion near the base of heavy-chain complementarity-determining region (CDR) H2 resulted in a bulge in that loop. This enlarged CDR H2 loop impinges on adjacent regions, causing distortion of the CDR H1 architecture and its displacement away from the antigen-combining site. Removal of the insertion restores the canonical structure of CDR H1 and CDR H2, but binding, neutralization activity, and in vivo activity were reduced markedly because of steric conflict of CDR H1 with the hemagglutinin antigen. PMID:21304166

  8. PRODUCTION AND CHARACTERIZATION OF MONOCLONAL ANTIBODIES, APTAMERS AND SINGLE CHAIN ANTIBODIES TO MYCOBACTERIUM AVIUM SUBSP. PARATUBERCULOSIS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Paratuberculosis (MAP) was identified as an unmet need at the 7th International Colloquium on Paratuberculosis in Bilbao, Spain. To fill this gap in Johne’s disease research, monoclonal antibodies (mAbs) against MAP were produced from BALB/c mice immunized with sonicated MAP extracts or recombinant...

  9. Novel antitenascin antibody with increased tumour localisation for Pretargeted Antibody-Guided RadioImmunoTherapy (PAGRIT)

    Microsoft Academic Search

    R De Santis; A M Anastasi; V D'Alessio; A Pelliccia; C Albertoni; A Rosi; B Leoni; R Lindstedt; F Petronzelli; M Dani; A Verdoliva; A Ippolito; N Campanile; V Manfredi; A Esposito; G Cassani; M Chinol; G Paganelli; P Carminati

    2003-01-01

    The Pretargeted Antibody-Guided RadioImmunoTherapy (PAGRIT) method is based on intravenous, sequential administration of a biotinylated antibody, avidin\\/streptavidin and 90Y-labelled biotin. The hybridoma clone producing the monoclonal antitenascin antibody BC4, previously used for clinical applications, was found not suitable for further development because of the production of an additional, nonfunctional light chain. In order to solve this problem, the new cST2146

  10. Modification of monoclonal antibody carbohydrates by oxidation, conjugation, or deoxymannojirimycin does not interfere with antibody effector functions

    Microsoft Academic Search

    Michel Awwad; Phoebe G. Strome; Steven C. Gilman; Helena R. Axelrod

    1994-01-01

    Site-specific attachment of metal chelators or cytotoxic agents to the carbohydrate region of monoclonal antibodies results in clinically useful immunoconjugates [Doerr et al. (1991) Ann Surg 214: 118, Wynant et al. (1991) Prostate 18: 229]. Since the capacity of monoclonal antibodies (mAb) to mediate tumor cell lysis via antibody-dependent cellular cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC) may accentuate the therapeutic

  11. Double-antibody sandwich ELISA using biotinylated antibodies for the detection of Echinococcus granulosus coproantigens in dogs

    Microsoft Academic Search

    Aitziber Benito; David Carmena

    2005-01-01

    Here we present the diagnostic evaluation of an improved double-antibody sandwich ELISA for detecting Echinococcus granulosus antigens in dog faecal samples (coproantigens). A purified rabbit IgG fraction against protoscolex excretory–secretory products was used as primary antibody, and the same fraction conjugated with biotin as secondary antibody. In order to validate the sandwich ELISA, intra- and inter-assay precision, linearity, and recovery

  12. Conversion of murine antibodies to human antibodies and their optimization for ovarian cancer therapy targeted to the folate receptor

    Microsoft Academic Search

    Mariangela Figini; Franck Martin; Renata Ferri; Elena Luison; Elena Ripamonti; Alberto Zacchetti; Mimosa Mortarino; Vito Di Cioccio; Giovanni Maurizi; Marcello Allegretti; Silvana Canevari

    2009-01-01

    We previously developed murine and chimeric antibodies against a specific epithelial ovarian carcinoma (EOC) marker, named\\u000a folate receptor (FR), and promising results were obtained in phase II trials. More recently, we successfully generated a completely\\u000a human Fab fragment, C4, by conversion of one of the murine anti-FR antibodies to human antibody using phage display and guided selection.\\u000a However, subsequent efforts

  13. Enzyme-Linked Immunosorbent Assay Using Glycoprotein and Monoclonal Antibody for Detecting Antibodies to Vesicular Stomatitis Virus Serotype New Jersey

    Microsoft Academic Search

    Hyang-Sim Lee; Eun-Jeong Heo; Hye-Young Jeoung; Hyo-Rim Ko; Chang-Hee Kweon; Hee-Jeong Youn; Young-Joon Ko

    2009-01-01

    In this study, an enzyme-linked immunosorbent assay (ELISA) using glycoprotein and a monoclonal antibody (MAb) was developed for the detection of antibodies to vesicular stomatitis virus (VSV) serotype New Jersey (NJ). The glycoprotein to be used as a diagnostic antigen was extracted from partially purified VSV-NJ, and a neutralizing MAb specific to VSV-NJ was incorporated to compete with antibodies in

  14. Fine specificity of anti-citrullinated peptide antibodies discloses a heterogeneous antibody population in rheumatoid arthritis

    PubMed Central

    Goules, J D; Goules, A V; Tzioufas, A G

    2013-01-01

    Anti-citrullinated peptide antibodies (ACPA) are highly specific for rheumatoid arthritis (RA). However, the predominant B cell epitopes have not yet been defined. The aim of this study was to examine the reactivity of ACPA against different peptides derived from citrullinated proteins and to investigate whether or not these antibodies constitute a homogeneous population. For this purpose, sera from patients with RA (n = 141), systemic lupus erythematosus (SLE) (n = 60), Sjögren's syndrome (SS) (n = 54) and healthy controls (n = 100) were tested for their reactivity against six citrullinated peptides derived from peptidyl arginine deiminase (PAD), vimentin (vim), alpha-enolase (enol), fibrin, type II collagen (col-II) and filaggrin, respectively. A non-citrullinated control peptide derived from PAD was used as control (ctrlPAD621–40). Antibody reactivity against each individual peptide was evaluated by enzyme-linked immunosorbent assay (ELISA). Specificity and cross-reactivity of ACPA were tested by using two prototype sera with homologous and cross-inhibition assays. Specificity of ACPA from two prototype sera was confirmed by purification of anti-peptide antibodies and homologous-inhibition experiments. We found that sera from patients with RA reacted diversely with the six citrullinated peptides. More specifically, PAD211–30 displayed 29·08% sensitivity, vim60–75 29·08%, enol5–21 37·59%, fibrin617–31 31·21%, col-II358–75 29·97% and filaggrin306–24 28·37%, while control ctrlPAD621–40 showed no reactivity. All reactive peptides were found to be highly specific for RA. A notable cross-reaction (>70%) was found mainly between filaggrin and the majority of anti-citrullinated peptide antibodies. We concluded that ACPA in RA constitute a heterogeneous population with limited cross-reactivity and without a predominant epitope. PMID:23711220

  15. Fine specificity of anti-citrullinated peptide antibodies discloses a heterogeneous antibody population in rheumatoid arthritis.

    PubMed

    Goules, J D; Goules, A V; Tzioufas, A G

    2013-10-01

    Anti-citrullinated peptide antibodies (ACPA) are highly specific for rheumatoid arthritis (RA). However, the predominant B cell epitopes have not yet been defined. The aim of this study was to examine the reactivity of ACPA against different peptides derived from citrullinated proteins and to investigate whether or not these antibodies constitute a homogeneous population. For this purpose, sera from patients with RA (n = 141), systemic lupus erythematosus (SLE) (n = 60), Sjögren's syndrome (SS) (n = 54) and healthy controls (n = 100) were tested for their reactivity against six citrullinated peptides derived from peptidyl arginine deiminase (PAD), vimentin (vim), alpha-enolase (enol), fibrin, type II collagen (col-II) and filaggrin, respectively. A non-citrullinated control peptide derived from PAD was used as control (ctrlPAD(621-40)). Antibody reactivity against each individual peptide was evaluated by enzyme-linked immunosorbent assay (ELISA). Specificity and cross-reactivity of ACPA were tested by using two prototype sera with homologous and cross-inhibition assays. Specificity of ACPA from two prototype sera was confirmed by purification of anti-peptide antibodies and homologous-inhibition experiments. We found that sera from patients with RA reacted diversely with the six citrullinated peptides. More specifically, PAD(211-30) displayed 29·08% sensitivity, vim(60-75) 29·08%, enol(5-21) 37·59%, fibrin(617-31) 31·21%, col-II(358-75) 29·97% and filaggrin(306-24) 28·37%, while control ctrlPAD(621-40) showed no reactivity. All reactive peptides were found to be highly specific for RA. A notable cross-reaction (>70%) was found mainly between filaggrin and the majority of anti-citrullinated peptide antibodies. We concluded that ACPA in RA constitute a heterogeneous population with limited cross-reactivity and without a predominant epitope. PMID:23711220

  16. Fully human monoclonal antibodies from antibody secreting cells after vaccination with Pneumovax®23 are serotype specific and facilitate opsonophagocytosis.

    PubMed

    Smith, Kenneth; Muther, Jennifer J; Duke, Angie L; McKee, Emily; Zheng, Nai-Ying; Wilson, Patrick C; James, Judith A

    2013-05-01

    B lymphocyte memory generates antibody-secreting cells (ASCs) that represent a source of protective antibodies that may be exploited for therapeutics. Here we vaccinated four donors with Pneumovax®23 and produced human monoclonal antibodies (hmAbs) from ASCs. We have cloned 137 hmAbs and the specificities of these antibodies encompass 19 of the 23 serotypes in the vaccine, as well as cell wall polysaccharide (CWPS). Although the majority of the antibodies are serotype specific, 12% cross-react with two serotypes. The Pneumovax®23 ASC antibody sequences are highly mutated and clonal, indicating an anamnestic response, even though this was a primary vaccination. Hmabs from 64% of the clonal families facilitate opsonophagocytosis. Although 9% of the total antibodies bind to CWPS impurity in the vaccine, none of these clonal families showed opsonophagocytic activity. Overall, these studies have allowed us to address unanswered questions in the field of human immune responses to polysaccharide vaccines, including the cross-reactivity of individual antibodies between serotypes and the percentage of antibodies that are protective after vaccination with Pneumovax®23. PMID:23084371

  17. Kinetics of intralymphatically delivered monoclonal antibodies

    SciTech Connect

    Wahl, R.L.; Geatti, O.; Liebert, M.; Beers, B.; Jackson, G.; Laino, L.; Kronberg, S.; Wilson, B.S.; Beierwaltes, W.H.

    1985-05-01

    Radiolabeled monoclonal antibody (MoAb) administration subcutaneously (sq), so that preferential uptake is to the lymphatics, holds significant promise for the detection of lymph node metastases. Only limited information is available about clearance rates of intralymphatically administered MoAbs. I-131 labeled intact IgG (225.28S), F(ab's)2 (225.28S) or IgM (FT162) were administered sq to anesthetized Balb/C mice. Eight mice were studied with each MoAb, 4 with a foot-pad injection, 4 with an anterior abdominal injection. Gamma camera images were collected into a computer, over the first 6 hrs after injection with the animals anesthetized and immobile. Animals were then allowed to move about freely. Additional images were then acquired out to 48 hrs. Regions of interest wre selected over the injection site and the kinetics of antibody egress determined. Clearance rates from local sq injection sites are influenced by motion and somewhat by location. The class and fragment status of the MoAb appear relatively less important in determining clearance rates from sq injections than they are in determining whole-body clearance after iv injections. Additional studies using Fab fragments and additional monoclonals will be useful in extending these observations.

  18. Mechanisms of action of CD20 antibodies

    PubMed Central

    Boross, Peter; Leusen, Jeanette H W

    2012-01-01

    Therapeutic monoclonal antibodies (mAbs) that target the CD20 antigen on B cells are successfully used in the clinic for the depletion of B cells to treat various forms of cancer and autoimmune diseases. The first CD20 mAb, approved by the FDA in 1998, was rituximab (RTX) and since then it has been widely used to treat more than one million patients thus far. The success of RTX has led to a general interest in the mechanism of action of CD20 mAbs. CD20 mAbs can induce tumor killing via various mechanisms, such as direct induction of apoptosis, antibody-dependent cell-mediated cytotoxicity (ADCC) or complement-dependent lysis (CDC). Although we now understand these mechanisms better, it is still unclear which of these mechanisms is the most important for in vivo RTX action. Not every patient respond to RTX treatment and eventually the overwhelming majority will experience a relapse. Therefore, there is an urgent need to improve the efficacy of CD20 mAbs. This review aims to summarize our current understanding on the mechanism of action of CD20 mAbs. PMID:23226614

  19. Hemostatic properties of a TFPI antibody.

    PubMed

    Petersen, Lars C

    2012-05-01

    Bleeds in hemophilia are treated either on demand or prophylactically by intravenous replacement therapy with FVIII or FIX. However, there is a call for subcutaneous and less frequent drug administration, and this need may be met by administration of a suitable antibody. Pioneering studies in vitro] and in a rabbit hemophilia model suggest that blockage of tissue factor pathway inhibitor (TFPI) provides a potential alternative approach to current therapy of hemophilia patients. TFPI down-regulates the initiation of coagulation by inhibition of FVIIa/TF/FXa and blockage of TFPI enhances FXa and thrombin generation. In line with these discoveries, TFPI targeting reagents with different potential benefits are currently evaluated as possible novel therapeutic agents. The development and testing of these agents in in vitro and in vivo hemophilia models provide new information about the mode of action of TFPI and its role in hemostasis. Blockage of TFPI with various antagonists has been shown to effectively enhance FX activation by TF/FVIIa and improve clot formation in hemophilia blood and plasma. The monoclonal antibody, mAb 2021, is one such antagonist directed towards the Kunitz-type protease inhibitor (KPI) 2 domain of TFPI which is now being tested in preclinical and clinical trials. Using mAb 2021, we have confirmed the original findings, and further characterized the pro-haemostatic effect of this specific anti-KPI-2 mAb in preclinical studies. PMID:22405586

  20. Seropositivity of Dengue Antibodies during Pregnancy

    PubMed Central

    Mohamed Ismail, Nor Azlin; Wan Abd Rahim, Wan Elly Rushima; Salleh, Sharifah Azura; Neoh, Hui-Min; Jamal, Rahman; Jamil, Muhammad Abdul

    2014-01-01

    Purpose. Malaysia a dengue endemic country with dengue infections in pregnancy on the rise. The present study was aimed at determining dengue seroprevalence (IgG or IgM) during pregnancy and its neonatal transmission in dengue seropositive women. Methods. Maternal with paired cord blood samples were tested for dengue antibodies (IgG and IgM) using an enzyme-linked immunosorbent assay (ELISA). Maternal age, parity, occupation, ethnic group, and gestational age were recorded. Data on neonatal Apgar score and admissions to the Neonatal Intensive Care Unit (NICU) were analyzed. Results. Out of 358 women recruited, about 128 (35.8%) patients were seropositive. Twelve patients (3.4%) had recent infections (IgM positive) and another 116 women (32.4%) were with past infections (IgG positive). All babies born to seropositive mothers had positive IgG paired cord blood; however, no IgM seropositivity was observed. All neonates had good Apgar scores and did not require NICU admission. Conclusion. In this study, 35.8% pregnant women were found to be dengue seropositive. However, transplacental transfer of IgG antibodies had no detrimental effect on the neonatal outcomes. PMID:25587564

  1. Immunofluorescent antibody test for diagnosis of gonorrhoea.

    PubMed Central

    Caloenescu, M; Clecner, B; Petrow, S; Kasatiya, S S

    1975-01-01

    An indirect fluorescent antibody test was evaluated in 198 cases of a high-risk group with a culture prevalence of 37.3% and in 426 cases of a low-risk group with a culture prevalence of 1.16%. A sensitivity of 77.1% in the culture-positive patients with uncomplicated gonorrhoea, and a specificity of 88.7% in the culture- and history-negative cases, was obtained in the high-risk group. In this group, the sera from 88.8% of the patients with culture-proven gonorrhoea became positive in an indirect fluorescent antibody test within 3 weeks of last sexual contact. In the low-risk group, for which the sensitivity could not be determined due to various reasons, a specificity of 95.8% was obtained. Complement fixation test was positive in sera of only 17.6% of the culture-positive cases of the high-risk group. PMID:809468

  2. Idiotypic specificity of rabbit antibodies to streptococcal group polysaccharides.

    PubMed

    Braun, D G; Kelus, A S

    1973-11-01

    Anti-idiotypic antisera against six restricted rabbit streptococcal group specific antibodies have been raised in rabbits matched for allotypes. All these antisera reacted specifically with their homologous idiotypes on double-diffusion tests in agarose gel. In addition, they showed a high incidence of cross-specificities with group-specific hyperimmune sera induced in both closely related and unrelated individuals. These precipitating cross-specificities could be explained for two systems by the interference of rheumatoid factor. Two idiotypic antibody systems have been analyzed in detail; these were restricted antibodies produced in a father and in one of his offspring. The methods employed included binding inhibition of radio-labeled homologous Fab fragments and hemagglutination inhibition with homologous idiotypic coat. The data demonstrated that only related rabbits produced, besides non-cross-reacting antibodies, idiotypically similar antibodies raised to the same antigen. About one-third of the cross-reactive idiotypes showed binding inhibition between 31 and 92%. Inhibition of binding above 50% in the paternal idiotypic system was only achieved by one offspring antibody whereas the F(1) progeny idiotypic system was inhibited to this extent by seven antibodies of related rabbits. In contrast, 87.5% and 91.7% of antibodies of unrelated rabbits were less than 20% inhibitory. Within this study two idiotypically identical antibodies have not been found. This implies that A-variant-specific antibodies of related rabbits which produced antipolysaccharide antibodies were structurally different. Cross-reaction, even if greater than 90% by binding inhibition, appears to involve only part and not all of the variable regions. PMID:4200777

  3. Antibodies to GABAA receptor ?1 and ?2 subunits

    PubMed Central

    Pettingill, Philippa; Kramer, Holger B.; Coebergh, Jan Adriaan; Pettingill, Rosie; Maxwell, Susan; Nibber, Anjan; Malaspina, Andrea; Jacob, Anu; Irani, Sarosh R.; Buckley, Camilla; Beeson, David; Lang, Bethan; Waters, Patrick

    2015-01-01

    Objective: To search for antibodies against neuronal cell surface proteins. Methods: Using immunoprecipitation from neuronal cultures and tandem mass spectrometry, we identified antibodies against the ?1 subunit of the ?-aminobutyric acid A receptor (GABAAR) in a patient whose immunoglobulin G (IgG) antibodies bound to hippocampal neurons. We searched 2,548 sera for antibodies binding to GABAAR ?, ?, and ? subunits on live HEK293 cells and identified the class, subclass, and GABAAR subunit specificities of the positive samples. Results: GABAAR-Abs were identified in 40 of 2,046 (2%) referred sera previously found negative for neuronal antibodies, in 5/502 (1%) previously positive for other neuronal surface antibodies, but not in 92 healthy individuals. The antibodies in 40% bound to either the ?1 (9/45, 20%) or the ?2 subunits (9/45, 20%) and were of IgG1 (94%) or IgG3 (6%) subclass. The remaining 60% had lower antibody titers (p = 0.0005), which were mainly immunoglobulin M (IgM) (p = 0.0025), and showed no defined subunit specificity. Incubation of primary hippocampal neurons with GABAAR IgG1 sera reduced surface GABAAR membrane expression. The clinical features of 15 patients (GABAAR ?1 n = 6, ?2 n = 5, undefined n = 4) included seizures (47%), memory impairment (47%), hallucinations (33%), or anxiety (20%). Most patients had not been given immunotherapies, but one with new-onset treatment-resistant catatonia made substantial improvement after plasma exchange. Conclusions: The GABAAR ?1 and ?2 are new targets for antibodies in autoimmune neurologic disease. The full spectrum of clinical features, treatment responses, correlation with antibody specificity, and in particular the role of the IgM antibodies will need to be assessed in future studies. PMID:25636713

  4. Role for antibodies in altering behavior and movement.

    PubMed

    Libbey, Jane E; Fujinami, Robert S

    2010-08-01

    At the past meeting of INSAR, the role of autoimmunity was discussed in an educational session. This article summarizes this discussion. In immune-mediated diseases, antibodies can contribute to the pathogenesis of the disease and are sometimes the force that drives the disease process. This concept has not been established for autism. In autoimmune diseases, such as systemic lupus erythematosus (SLE), antibodies are found to react with double-stranded DNA. These antibodies also cross-react with N-methyl-D aspartate receptors. Many SLE patients suffer neurologic syndromes of the central nervous system (CNS). Similarly individuals infected with Group A streptococcus (GAS) have antibodies against the GAS carbohydrate, which cross-react with tubulin and lysoganglioside GM1 on neurons. During the acute stage of infection, GAS-infected patients develop Syndenham chorea where the disease process is driven in part by these cross-reactive antibodies. As the antibody levels decrease, the clinical features of Syndenham chorea resolve. In these two immune-mediated diseases, antibodies clearly play a role in the pathogenesis of the diseases. There are reports that mothers of individuals with autism have antibodies that react with brain proteins and when these antibodies are passively transferred to pregnant non-human primates or rodents the offspring has behavioral and nervous system changes. It is still not clear whether the antibodies found in mothers of individuals with autism actually play a role in the disease. More studies need to be performed to identify the proteins recognized by the antibodies and to determine how these could affect development, behavior and changes within the CNS. PMID:20589715

  5. 21 CFR 660.1 - Antibody to Hepatitis B Surface Antigen.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 2013-04-01 false Antibody to Hepatitis B Surface Antigen. 660.1 Section...SUBSTANCES FOR LABORATORY TESTS Antibody to Hepatitis B Surface Antigen § 660.1 Antibody to Hepatitis B Surface Antigen. (a) Proper...

  6. Monoclonal Antibody Epitope Mapping Describes Tailspike -Helix Folding and Aggregation Intermediates*S

    E-print Network

    Clark, Patricia L.

    Monoclonal Antibody Epitope Mapping Describes Tailspike -Helix Folding and Aggregation, nine -tailspike antibody binding epitopes were characterized by measuring the binding of these mono that the antibody epitopes are distributed throughout the tailspike struc- ture, with several clustered

  7. 21 CFR 660.1 - Antibody to Hepatitis B Surface Antigen.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 2010-04-01 false Antibody to Hepatitis B Surface Antigen. 660.1 Section...SUBSTANCES FOR LABORATORY TESTS Antibody to Hepatitis B Surface Antigen § 660.1 Antibody to Hepatitis B Surface Antigen. (a) Proper...

  8. 21 CFR 864.9175 - Automated blood grouping and antibody test system.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...antibody test system. (a) Identification. An automated blood grouping and antibody test system is a device used to group erythrocytes (red blood cells) and to detect antibodies to blood group antigens. (b) Classification. Class II...

  9. 21 CFR 660.1 - Antibody to Hepatitis B Surface Antigen.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 2012-04-01 false Antibody to Hepatitis B Surface Antigen. 660.1 Section...SUBSTANCES FOR LABORATORY TESTS Antibody to Hepatitis B Surface Antigen § 660.1 Antibody to Hepatitis B Surface Antigen. (a) Proper...

  10. 21 CFR 660.1 - Antibody to Hepatitis B Surface Antigen.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 2014-04-01 false Antibody to Hepatitis B Surface Antigen. 660.1 Section...SUBSTANCES FOR LABORATORY TESTS Antibody to Hepatitis B Surface Antigen § 660.1 Antibody to Hepatitis B Surface Antigen. (a) Proper...

  11. Antibodies production and the maintenance of the immunological memory

    NASA Astrophysics Data System (ADS)

    de Castro, A.

    2006-02-01

    In this work we have considered the R. Nayak et al. [Immunology 102, 387 (2001)] biological model - Relay Hypothesis - to study the time evolution of the clonal repertoire, including the populations of antibodies. Our results suggest that the decrease of the production of antibodies favors the global maintenance of immune memory.

  12. Evaluation of Recombinant Antibodies on Protein Microarrays Applying the Multiple

    E-print Network

    Konthur, Zoltán

    that all 96 parallel selections resulted in polyclonal enrichment of phage particles and that for each-automated concepts have been introduced for the selection of recombinant binders from combinatorial phage display antibody libraries. These include the parallel selection of antibody-displaying phage mole- cules

  13. The significance of antibody coated bacteria in neuropathic bladder urines

    Microsoft Academic Search

    Rosemary Lindan

    1981-01-01

    A total of 234 patients with neuropathic bladder dysfunction and bacteria in the urine have been studied for the presence of antibody coating on the bacteria. Approximately one third of the patients so studied were found to have antibody coated bacteria in the urine (ACB + ) by fluorescent microscopy. No correlation could be found between evidence of active tissue

  14. Human-like antibodies neutralizing Western equine encephalitis virus

    PubMed Central

    Hülseweh, Birgit; Rülker, Torsten; Pelat, Thibaut; Langermann, Claudia; Frenzel, Andrč; Schirrmann, Thomas; Dübel, Stefan; Thullier, Philippe; Hust, Michael

    2014-01-01

    This study describes the development of the first neutralizing antibodies against Western equine encephalitis virus (WEEV), a member of the genus Alphavirus. WEEV is transmitted by mosquitoes and can spread to the human central nervous system, causing symptoms ranging from mild febrile reactions to life-threatening encephalitis. WEEV has been classified as a biological warfare agent by the US Centers for Disease Control and Prevention. No anti-WEEV drugs are currently commercially available. Neutralizing antibodies are useful for the pre- and post-exposure treatment of WEEV infections. In this study, two immune antibody gene libraries were constructed from two macaques immunized with inactivated WEEV. Four antibodies were selected from these libraries and recloned as scFv-Fc, with a human Fc part. These antibodies bound WEEV specifically in ELISA with little or no cross-reaction with other alphaviruses. They were further analyzed by immunohistochemistry. All binders were suitable for the intracellular detection of WEEV particles. Neutralizing activity was determined in vitro. Three of the four antibodies were found to be neutralizing; about 1 ng/mL of the best antibody (ToR69–3A2) neutralized 50% of 5x104 TCID50/mL. Due to its human-like nature with a germinality index of 89% (VH) and 91% (VL), the ToR69–3A2 antibody is a promising candidate for future passive vaccine development. PMID:24518197

  15. 2005 Nature Publishing Group Antibody-based therapies for malaria

    E-print Network

    Cai, Long

    © 2005 Nature Publishing Group OPI N ION Antibody-based therapies for malaria Richard J. Pleass and tissue fluids, and can protect against malaria by binding and neutralizing malaria parasites of antibodies in the treatment and prevention of malaria. Despite control efforts, malaria continues to kill ~2

  16. Palladium-109 labeled anti-melanoma monoclonal antibodies

    DOEpatents

    Srivastava, S.C.; Fawwaz, R.A.; Ferrone, S.

    1984-04-30

    The invention consists of new monoclonal antibodies labelled with Palladium 109, a beta-emitting radionuclide, the method of preparing this material, and its use in the radiotherapy of melanoma. The antibodies are chelate-conjugated and demonstrate a high uptake in melanomas. (ACR)

  17. RESEARCH ARTICLE Detection of pancreatic cancer using antibody

    E-print Network

    Peterson, Carsten

    RESEARCH ARTICLE Detection of pancreatic cancer using antibody microarray-based serum proteinFv antibody microarray in an attempt to classify sera derived from pancreatic adenocarcinoma patients versus cancer patients and healthy subjects. Furthermore, a potential protein signature, consisting of 21

  18. The Characterization of Monoclonal Antibodies to Newcastle Disease Virus

    Microsoft Academic Search

    P. H. Russell; P. C. Griffiths; K. K. A. Goswami; D. J. Alexander; M. J. Cannon; W. C. Russell

    1983-01-01

    SUMMARY Monoclonal antibodies to the haemagglutinin-neuraminidase (HN), fusion (F), polymerase and nucleocapsid polypeptides of Newcastle disease virus were prepared. Two epitopes were recognized on the HN polypeptide: one was associated with inhibition of haemagglutination and poor neutralization and the other with good neutralization and no inhibition of haemagglutination. The most effective neutralizing antibody was that produced against the F polypeptide.

  19. SolidPhase Radioimmunoassay in Antibody-Coated Tubes

    Microsoft Academic Search

    Kevin Catt; Geoffrey W. Tregear

    1967-01-01

    The adsorption of antibody to polymeric surfaces has been used to develop a new method of solid-phase radioimmunoassay. Incubation is performed in antibody-coated, disposable tubes that are finally washed-out with water and counted for quantitation of the bound tracer. The method is simple, rapid, inexpensive, and suitable for automation.

  20. Monoclonal antibodies reactive with mucin expressed in breast cancer

    Microsoft Academic Search

    P-X Xing; JJ Tjandra; SA Stacker; JG Teh; CH Thompson; PJ McLaughlin; IFC McKenzie

    1989-01-01

    Three murine monoclonal antibodies (BC 1, BC2 and BC3) were developed against human milk fat globule membrane (HMFGM). By immunoperoxidase staining, it was found that the antigenic determinants had a predominant distribution in breast cancer tissue. In addition, the antibodies reacted preferentially with mucin derived from human milk rather than that derived from the breast cancer cell line ZR75; they

  1. Evanescent wave immunoprobe with high bivalent antibody activity

    Microsoft Academic Search

    Sandra F. Feldman; Egidijus E. Uzgiris; C. Murray Penney; John Y. Gui; Emily Y. Shu; Edward B. Stokes

    1995-01-01

    We have determined the kinetic response of an evanescent wave optical fibre immunosensor and its absolute sensitivity. Using both kinetic methods and optical determinations of bound antigen, we infer that there are 2·4 × 1011 active antibodies per cm2 probe area. We estimate that 75% of the active antibodies are in the bivalent form, with both binding site capable of

  2. The role of phage display in therapeutic antibody discovery.

    PubMed

    Chan, Conrad E Z; Lim, Angeline P C; MacAry, Paul A; Hanson, Brendon J

    2014-12-01

    Phage display involves the expression of selected proteins on the surface of filamentous phage through fusion with phage coat protein, with the genetic sequence packaged within, linking phenotype to genotype selection. When combined with antibody libraries, phage display allows for rapid in vitro selection of antigen-specific antibodies and recovery of their corresponding coding sequence. Large non-immune and synthetic human libraries have been constructed as well as smaller immune libraries based on capturing a single individual's immune repertoire. This completely in vitro process allows for isolation of antibodies against poorly immunogenic targets as well as those that cannot be obtained by animal immunization, thus further expanding the utility of the approach. Phage antibody display represents the first developed methodology for high throughput screening for human therapeutic antibody candidates. Recently, other methods have been developed for generation of fully human therapeutic antibodies, such as single B-cell screening, next-generation genome sequencing and transgenic mice with human germline B-cell genes. While each of these have their particular advantages, phage display has remained a key methodology for human antibody discovery due its in vitro process. Here, we review the continuing role of this technique alongside other developing technologies for therapeutic antibody discovery. PMID:25135889

  3. MONOCLONAL ANTIBODY TO FENBENDAZOLE: UTILITY IN RESIDUE STUDIES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A monoclonal antibody-based ELISA was developed for fenbendazole, a widely used benzimidazole anthelmintic, with approved uses in cattle and other food animals. The antibody was elicited using as hapten 2-succinamido-5(6)-phenylthiobenzimidazole, which was conjugated with bovine serum albumin to pro...

  4. The development of potential antibody-based therapies for myeloma

    PubMed Central

    Sherbenou, Daniel W.; Behrens, Christopher R.; Su, Yang; Wolf, Jeffrey L.; Martin, Thomas G.; Liu, Bin

    2015-01-01

    With optimal target antigen selection antibody-based therapeutics can be very effective agents for hematologic malignancies, but none have yet been approved for myeloma. Rituximab and brentuximab vedotin are examples of success for the naked antibody and antibody–drug conjugate classes, respectively. Plasma cell myeloma is an attractive disease for antibody-based targeting due to target cell accessibility and the complementary mechanism of action with approved therapies. Initial antibodies tested in myeloma were disappointing. However, recent results from targeting well-characterized antigens have been more encouraging. In particular, the CD38 and CD138 targeted therapies are showing single-agent activity in early phase clinical trials. Here we will review the development pipeline for naked antibodies and antibody–drug conjugates for myeloma. There is clear clinical need for new treatments, as myeloma inevitably becomes refractory to standard agents. The full impact is yet to be established, but we are optimistic that the first FDA-approved antibody therapeutic(s) for this disease will emerge in the near future. PMID:25294123

  5. Radioimmunodetection of human melanoma tumor xenografts with human monoclonal antibodies.

    PubMed

    Gomibuchi, M; Saxton, R E; Lake, R R; Katano, M; Irie, R F

    1986-01-01

    We established a human IgM monoclonal antibody that defines a tumor-associated membrane antigen expressed on human melanoma cells. The antigen has been identified as the ganglioside GD2. In this paper, we describe the potential usefulness of the human monoclonal antibody for radioimaging. Nude mice bearing tumors derived from a human melanoma cell line were used as a model. Antibody activity was degradated significantly after labeling with 131I by the use of a modified chloramine-T method. After testing various concentrations, labeled antibody of a specific activity of 2.8 microCi/micrograms produced the best results. Balb/c nude mice bearing a GD2-positive M14 melanoma cell line were injected with 10-30 micrograms of labeled antibody, and its radiolocalization in different organs and in the whole body were evaluated. The best tumor image was obtained on Day 6. The labeled antibody uptake ratio between tumor and muscle was 9.2:1; the ratio between tumor and liver was 1.4:1. These studies represent the first report of experimental tumor imaging with human monoclonal antibody. Human monoclonals with probably prove to be superior reagents for tumor imaging in melanoma patients if the problem of antibody radiolysis is resolved. PMID:3771234

  6. PRODUCTION OF RECOMBINANT ANTIBODIES FOR LOW-ABUNDANCE PROTEINS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Production of high quality antibodies is extremely important for the detection of low abundance proteins. Tristetraprolin (TTP), an anti-inflammatory protein, is such a very low-abundance protein in normal cells and tissues. Numerous laboratories and a few companies have produced TTP antibodies, but...

  7. Cellulose antibody films for highly specific evanescent wave immunosensors

    Microsoft Academic Search

    A. Hartmann; D. Bock; T. Jaworek; S. Kaul; M. Schulze; H. Tebbe; Gerhard Wegner; Stefan Seeger

    1996-01-01

    For the production of recognition elements for evanescent wave immunosensors optical waveguides have to be coated with ultrathin stable antibody films. In the present work non amphiphilic alkylated cellulose and copolyglutamate films are tested as monolayer matrices for the antibody immobilization using the Langmuir-Blodgett technique. These films are transferred onto optical waveguides and serve as excellent matrices for the immobilization

  8. Antibody secreting cell assay for influenza A virus in swine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An ELISPOT assay to enumerate B-cells producing antibodies specific to a given antigen, also known as an antibody secreting cell (ASC) assay, was adapted to detect B-cells specific for influenza A virus (IAV). The assay is performed ex vivo and enumerates ASC at a single cell level. A simple ASC det...

  9. Diagnosis of infectious laryngotracheitis using a monoclonal antibody ELISA

    Microsoft Academic Search

    Jennifer J. York; K. J. Fahey

    1988-01-01

    An ELISA has been developed which uses a selected monoclonal antibody specific for ILT virus. The ELISA proved to be as accurate as, yet faster than, virus isolation, more accurate than the fluorescent antibody test and more accurate and rapid than the relatively simple agar gel precipitin test.The ELISA clearly differentiated between chickens from commercial flocks infected with ILT virus

  10. In situ characterization of antibody grafting on porous monolithic supports.

    PubMed

    Faye, C; Chamieh, J; Moreau, T; Granier, F; Faure, K; Dugas, V; Demesmay, C; Vandenabeele-Trambouze, O

    2012-01-15

    The efficient immobilization of antibodies on monolithic support is one of the most critical steps when preparing immunoaffinity supports. In this work, the ADECA (amino density estimation by colorimetric assay) method was adapted to tridimensional supports (in a dynamic mode) and proved to be efficient to characterize the antibodies grafting efficiency on 15.3±0.9mg porous glycidyl methacrylate (GMA)-co-ethylene dimethacrylate (EDMA) monolithic columns. The amount of grafted antibodies measured in situ on the monolith by ADECA (8.2±0.2?g of antibodies per milligram of monolith) was consistent with values obtained by bicinchoninic acid assay (BCA) after crushing the monolith. ADECA was shown to be less time-consuming and more versatile than BCA. The ADECA method was further implemented to thoroughly study and optimize the antibody grafting conditions (influence of pH and kinetics of the grafting step) on GMA-based monoliths and to check the covalent nature of the antibody/surface linking and its stability. Using the total amount of grafted antibodies and the amount of recognized antigen, we found that 65±6% of antibodies were able to capture their antigen. Finally, the grafting of Fab and F(ab')(2) fragments demonstrated that no significant improvement of the global binding capacity of the monolith was obtained. PMID:21982863

  11. Structural Comparison of Different Antibodies Interacting with Parvovirus Capsids

    SciTech Connect

    Hafenstein, Susan; Bowman, Valorie D.; Sun, Tao; Nelson, Christian D.S.; Palermo, Laura M.; Chipman, Paul R.; Battisti, Anthony J.; Parrish, Colin R.; Rossmann, Michael G.; Cornell; Purdue

    2009-05-13

    The structures of canine parvovirus (CPV) and feline parvovirus (FPV) complexed with antibody fragments from eight different neutralizing monoclonal antibodies were determined by cryo-electron microscopy (cryoEM) reconstruction to resolutions varying from 8.5 to 18 {angstrom}. The crystal structure of one of the Fab molecules and the sequence of the variable domain for each of the Fab molecules have been determined. The structures of Fab fragments not determined crystallographically were predicted by homology modeling according to the amino acid sequence. Fitting of the Fab and virus structures into the cryoEM densities identified the footprints of each antibody on the viral surface. As anticipated from earlier analyses, the Fab binding sites are directed to two epitopes, A and B. The A site is on an exposed part of the surface near an icosahedral threefold axis, whereas the B site is about equidistant from the surrounding five-, three-, and twofold axes. One antibody directed to the A site binds CPV but not FPV. Two of the antibodies directed to the B site neutralize the virus as Fab fragments. The differences in antibody properties have been linked to the amino acids within the antibody footprints, the position of the binding site relative to the icosahedral symmetry elements, and the orientation of the Fab structure relative to the surface of the virus. Most of the exposed surface area was antigenic, although each of the antibodies had a common area of overlap that coincided with the positions of the previously mapped escape mutations.

  12. Highly efficient transient expression of functional recombinant antibodies in lettuce

    Microsoft Academic Search

    Valentine Negrouk; Galina Eisner; Hyung-il Lee; Kaiping Han; Dean Taylor; Hing C. Wong

    2005-01-01

    An efficient, scalable transient expression system has been developed and characterized for production of recombinant antibodies in lettuce. The transient expression system utilizes lettuce (Lactuca sativa L.) vacuum-infiltrated with Agrobacterium tumefaciens bearing antibody genes cloned into a single or two separate expression vectors. The lettuce can be obtained commercially (grocery stores or directly from growers), so plant growth facilities are

  13. West Nile Virus Antibody Prevalence in Wild Mammals, Southern Wisconsin

    PubMed Central

    Docherty, Douglas E.; Nolden, Cherrie A.; Egstad, Kristina F.; Griffin, Kathryn M.

    2006-01-01

    Twenty percent prevalence of West Nile virus antibody was found in free-ranging medium-sized Wisconsin mammals. No significant differences were noted in antibody prevalence with regard to sex, age, month of collection, or species. Our results suggest a similar route of infection in these mammals. PMID:17326959

  14. Anti-LSP antibodies in acute liver disease

    Microsoft Academic Search

    R Meliconi; A Perperas; D Jensen; A Alberti; I G McFarlane; A L Eddleston; R Williams

    1982-01-01

    Sera from 71 patients with acute liver injury have been tested for antibodies to hepatocyte membrane lipoprotein complex (LSP) using a sensitive radioimmunoassay. Two main patterns of anti-LSP response were seen. In the first, seen in patients with type A and B viral hepatitis, anti-LSP antibodies were detectable at presentation, with the highest titres two to 10 days before the

  15. Antibody Profiles Characteristic of Mycobacterium tuberculosis Infection State

    Microsoft Academic Search

    Amy Davidow; Ganga V. Kanaujia; Lanbo Shi; Justin Kaviar; XuDong Guo; Nackmoon Sung; Gilla Kaplan; Dick Menzies; Maria L. Gennaro

    2005-01-01

    The relationship between specific antibody profiles and tuberculosis (TB) state was investigated by mea- suring serum antibody levels to six Mycobacterium tuberculosis antigens in human subjects grouped into four diagnostic categories: active disease, inactive (past) tuberculosis, latent infection without radiographic chest abnormalities, and infection free. Statistical data analyses showed that the latter two groups were serologically indistinguishable and that active

  16. SYNTHETIC PEPTIDES AND THEIR ANTIBODIES FOR MAPPING THE

    E-print Network

    Kasher, Roni

    SYNTHETIC PEPTIDES AND THEIR ANTIBODIES FOR MAPPING THE -BTX BINDING SITE Antibodies to short synthetic peptides from the -subunit of AChR were first employed in our lab for mapping the -BTX binding 192 and 193, contains the essential ele- ments for -BTX binding.6,8 This peptide binds - BTX directly

  17. Monoclonal Antibody Therapy for Relapsed or Treatment-Resistant Neuroblastoma

    Cancer.gov

    NCI is sponsoring two clinical trials of a monoclonal antibody called ch14.18, in combination with other drugs, to see if the antibody may be helpful for children or young adults (up to age 21) with relapsed or refractory high-risk neuroblastoma.

  18. Evaluation of inhibitor antibody in hemophiliaA population

    PubMed Central

    Mahmoodi Nesheli, Hassan; Hadizadeh, Amereh; Bijani, Ali

    2013-01-01

    Background: Inhibitor antibody to exogenous Factor VIII (FVIII) is a major complication of hemophilia treatment. This study was conducted to determine the prevalence of inhibitor antibody directed against FVIII. Methods: From May 2010 to May 2011, 52 patients with severe hemophilia A admitted in Amirkola Children’s Hospital were evaluated. Those who had abnormal mixing study, antibody against FVIII were measured. Data were collected and analyzed. Results: The age range of the patients was 4-60 years. The inhibitor antibody was seen in 9 (17.3%) patients. The mean age of patients with inhibitor at the time of diagnosis was 10.22 years (ranged 4-31 years). Old patients had more hemarthrosis than young patients. The mean level of inhibitor antibody was 8.47 Bethesda unit (ranged 2.3-29). Six patients had inhibitor antibody level ?5 Bethesda unit and three patients had inhibitor antibody level <5 Bethesda unit. Conclusion: This study showed that the prevalence of inhibitor antibodies in young patients is more than the old patients. PMID:24009969

  19. The development of potential antibody-based therapies for myeloma.

    PubMed

    Sherbenou, Daniel W; Behrens, Christopher R; Su, Yang; Wolf, Jeffrey L; Martin, Thomas G; Liu, Bin

    2015-03-01

    With optimal target antigen selection antibody-based therapeutics can be very effective agents for hematologic malignancies, but none have yet been approved for myeloma. Rituximab and brentuximab vedotin are examples of success for the naked antibody and antibody-drug conjugate classes, respectively. Plasma cell myeloma is an attractive disease for antibody-based targeting due to target cell accessibility and the complementary mechanism of action with approved therapies. Initial antibodies tested in myeloma were disappointing. However, recent results from targeting well-characterized antigens have been more encouraging. In particular, the CD38 and CD138 targeted therapies are showing single-agent activity in early phase clinical trials. Here we will review the development pipeline for naked antibodies and antibody-drug conjugates for myeloma. There is clear clinical need for new treatments, as myeloma inevitably becomes refractory to standard agents. The full impact is yet to be established, but we are optimistic that the first FDA-approved antibody therapeutic(s) for this disease will emerge in the near future. PMID:25294123

  20. Engineering anti-GD2 monoclonal antibodies for cancer immunotherapy.

    PubMed

    Ahmed, Mahiuddin; Cheung, Nai-Kong V

    2014-01-21

    Ganglioside GD2 is highly expressed on neuroectoderm-derived tumors and sarcomas, including neuroblastoma, retinoblastoma, melanoma, small cell lung cancer, brain tumors, osteosarcoma, rhabdomyosarcoma, Ewing's sarcoma in children and adolescents, as well as liposarcoma, fibrosarcoma, leiomyosarcoma and other soft tissue sarcomas in adults. Since GD2 expression in normal tissues is restricted to the brain, which is inaccessible to circulating antibodies, and in selected peripheral nerves and melanocytes, it was deemed a suitable target for systemic tumor immunotherapy. Anti-GD2 antibodies have been actively tested in clinical trials for neuroblastoma for over the past two decades, with proven safety and efficacy. The main limitations have been acute pain toxicity associated with GD2 expression on peripheral nerve fibers and the inability of antibodies to treat bulky tumor. Several strategies have been developed to reduce pain toxicity, including bypassing complement activation, using blocking antibodies, or targeting of O-acetyl-GD2 derivative that is not expressed on peripheral nerves. To enhance anti-tumor efficacy, anti-GD2 monoclonal antibodies and fragments have been engineered into immunocytokines, immunotoxins, antibody drug conjugates, radiolabeled antibodies, targeted nanoparticles, T-cell engaging bispecific antibodies, and chimeric antigen receptors. The challenges of these approaches will be reviewed to build a perspective for next generation anti-GD2 therapeutics in cancer therapy. PMID:24295643

  1. PREVALENCE OF ANTIBODIES TO ENCEPHALITOZOON CUNICULI IN HORSES IN BRAZIL.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Encephalitozoon cuniculi has been associated with natural cases of abortion and still-birth in horses. However, little is known abut the prevalence of this parasite in horses. We examined serva from 559 horses from Brazil for antibodies to E. cuniculi using the indirect immunofluorescent antibody ...

  2. SEROPREVALENCE OF NEOSPORA CANINUM ANTIBODIES IN DOGS IN INDIA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Neospora caninum is one of most important causes of abortion in cattle worldwide and dogs are an important risk factor for N. caninum infection in cattle. Antibodies to N. caninum were determined in 184 (126 rural, 58 urban) dogs from the Punjab State, India, by a commercial monoclonal antibody bas...

  3. Perfluorooctanoic Acid Exposure Suppresses T-independent Antibody Responses

    EPA Science Inventory

    Exposure to ż 3.75mg/kg of perfluoroocatnoic acid (PFOA) for 15d suppresses T-dependent antibody responses (TDAR), suggesting that T helper cells and/or B cells/plasma cells may be impacted. This study evaluated effects of PFOA exposure on the T cell-independent antibody response...

  4. Antibody Structure and the Generation of B-cell Diversity

    E-print Network

    Utrecht, Universiteit

    does an IgG molecule look like, what kind and how many chains? · What domains/region can and function of IgG and fragments · IgG1 · Fab & Fc · Antigen binding sites · Generation of antibodies therapeutic antibodies · EGFR as therapeutic target Agenda · Structure and function of IgG and fragments · IgG

  5. Immunomodulatory antibody therapy of cancer: the closer, the better.

    PubMed

    Dronca, Roxana S; Dong, Haidong

    2015-03-01

    Immune checkpoint blockade therapies have demonstrated promising therapeutic effects; however, clinical outcomes are variable, with only a subgroup of cancer patients achieving durable complete responses. New therapeutic strategies, including local administration of immunomodulatory antibodies, have been considered as better routes for improving the overall efficacy of antibody-based therapy. PMID:25351746

  6. Monitoring Monoclonal Antibody Delivery in Oncology: The Example of Bevacizumab

    E-print Network

    Paris-Sud XI, Université de

    Monitoring Monoclonal Antibody Delivery in Oncology: The Example of Bevacizumab Guillaume Nugue1 antibodies paves the way for new strategies in oncology using targeted therapy which should improve in Oncology: The Example of Bevacizumab. PLoS ONE 8(8): e72021. doi:10.1371/journal.pone.0072021 Editor

  7. Monoclonal antibodies: new agents for cancer detection and targeted therapy

    Microsoft Academic Search

    R. W. Baldwin; V. S. Byers

    1991-01-01

    Antibodies directed against markers on cancer cells are gaining in importance for the purpose of targeting diagnostic and therapeutic agents. In the past, this approach has had very limited success principally because the classical methods for producing antibodies from blood serum of animals immunized with cancer cells or extracts were unsatisfactory. The situation has changed dramatically since 1975 following the

  8. Antibody and B cell responses to Plasmodium sporozoites

    PubMed Central

    Dups, Johanna N.; Pepper, Marion; Cockburn, Ian A.

    2014-01-01

    Antibodies are capable of blocking infection of the liver by Plasmodium sporozoites. Accordingly the induction of anti-sporozoite antibodies is a major aim of various vaccine approaches to malaria. In recent years our knowledge of the specificity and quantities of antibodies required for protection has been greatly expanded by clinical trials of various whole sporozoite and subunit vaccines. Moreover, the development of humanized mouse models and transgenic parasites have also aided our ability to assess the specificity of antibodies and their ability to block infection. Nonetheless, considerable gaps remain in our knowledge – in particular in understanding what antigens are recognized by infection blocking antibodies and in knowing how we can induce robust, long-lived antibody responses. Maintaining high levels of circulating antibodies is likely to be of primary importance, as antibodies must block infection in the short time it takes for sporozoites to reach the liver from the skin. It is clear that a better understanding of the development of protective B cell-mediated immunity will aid the development and refinement of malaria vaccines. PMID:25477870

  9. Proteochemometric Modeling of the Antigen-Antibody Interaction: New Fingerprints for Antigen, Antibody and Epitope-Paratope Interaction

    PubMed Central

    Qiu, Tianyi; Xiao, Han; Zhang, Qingchen; Qiu, Jingxuan; Yang, Yiyan; Wu, Dingfeng; Cao, Zhiwei; Zhu, Ruixin

    2015-01-01

    Despite the high specificity between antigen and antibody binding, similar epitopes can be recognized or cross-neutralized by paratopes of antibody with different binding affinities. How to accurately characterize this slight variation which may or may not change the antigen-antibody binding affinity is a key issue in this area. In this report, by combining cylinder model with shell structure model, a new fingerprint was introduced to describe both the structural and physical-chemical features of the antigen and antibody protein. Furthermore, beside the description of individual protein, the specific epitope-paratope interaction fingerprint (EPIF) was developed to reflect the bond and the environment of the antigen-antibody interface. Finally, Proteochemometric Modeling of the antigen-antibody interaction was established and evaluated on 429 antigen-antibody complexes. By using only protein descriptors, our model achieved the best performance (R2=0.91,Qtest2=0.68) among peers. Further, together with EPIF as a new cross-term, our model (R2=0.92,Qtest2=0.74) can significantly outperform peers with multiplication of ligand and protein descriptors as a cross-term (R2?0.81,Qtest2?0.44). Results illustrated that: 1) our newly designed protein fingerprints and EPIF can better describe the antigen-antibody interaction; 2) EPIF is a better and specific cross-term in Proteochemometric Modeling for antigen-antibody interaction. The fingerprints designed in this study will provide assistance to the description of antigen-antibody binding, and in future, it may be valuable help for the high-throughput antibody screening. The algorithm is freely available on request. PMID:25901362

  10. Self-Assembled Antibody Multimers through Peptide Nucleic Acid Conjugation

    PubMed Central

    Kazane, Stephanie A.; Axup, Jun Y; Kim, Chan Hyuk; Ciobanu, Mihai; Wold, Erik D.; Barluenga, Sofia; Hutchins, Benjamin A.; Schultz, Peter G.; Winssinger, Nicolas; Smider, Vaughn V.

    2013-01-01

    With the recent clinical success of bispecific antibodies, a strategy to rapidly synthesize and evaluate bispecific or higher order multispecific molecules could facilitate the discovery of new therapeutic agents. Here we show that unnatural amino acids (UAAs) with orthogonal chemical reactivity can be used to generate site-specific antibody-oligonucleotide conjugates. These constructs can then be self-assembled into multimeric complexes with defined composition, valency and geometry. Using this approach, we generated potent bispecific antibodies that recruit cytotoxic T lymphocytes to Her2 and CD20 positive cancer cells, as well as multimeric antibody fragments with enhanced activity. This strategy should accelerate the synthesis and in vitro characterization of antibody constructs with unique specificities and molecular architectures. PMID:23210862

  11. Engineered antibody Fc variants with enhanced effector function

    NASA Astrophysics Data System (ADS)

    Lazar, Greg A.; Dang, Wei; Karki, Sher; Vafa, Omid; Peng, Judy S.; Hyun, Linus; Chan, Cheryl; Chung, Helen S.; Eivazi, Araz; Yoder, Sean C.; Vielmetter, Jost; Carmichael, David F.; Hayes, Robert J.; Dahiyat, Bassil I.

    2006-03-01

    Antibody-dependent cell-mediated cytotoxicity, a key effector function for the clinical efficacy of monoclonal antibodies, is mediated primarily through a set of closely related Fc receptors with both activating and inhibitory activities. By using computational design algorithms and high-throughput screening, we have engineered a series of Fc variants with optimized Fc receptor affinity and specificity. The designed variants display >2 orders of magnitude enhancement of in vitro effector function, enable efficacy against cells expressing low levels of target antigen, and result in increased cytotoxicity in an in vivo preclinical model. Our engineered Fc regions offer a means for improving the next generation of therapeutic antibodies and have the potential to broaden the diversity of antigens that can be targeted for antibody-based tumor therapy. antibody-dependent cell-mediated cytotoxicity | FcR | protein engineering | cancer

  12. Targeting of lymphocytes with 111In-labelled monoclonal antibodies.

    PubMed

    Loutfi, I; Batchelor, J R; Chisholm, P M; Epenetos, A A; Lavender, J P

    1988-10-01

    In this paper, we emphasize the rationale and work-up studies for using two radiolabelled anti-lymphocyte monoclonal antibodies for in vivo application as radiolabelling agents for T and B cells. In vitro experimental work involved radioimmunoassays on human lymphoid cell lines and anticoagulated whole blood with identification of relevant binding kinetics in terms of antibody internalization and elution. We tested also for the effect of the radiolabelled monoclonal antibodies on in vitro cell function defined as mitogen-induced proliferation in whole blood. As a final work-up in an animal model, the distribution of both unlabelled and 111In-labelled anti-Pan T cell monoclonal antibody was studied in the rat. Results from the in vitro experiments pointed to the possibility of using the described technique for specific lymphocyte radiolabelling. The in vivo application enabled us to identify optimal doses of antibody and radioactivity which showed agreement with the in vitro data. PMID:3211438

  13. Immunopotentiating properties of a multispecific ?-anti-idiotype antibody.

    PubMed

    Hernández, Tays; Mateo de Acosta, Cristina; Pérez, Rolando

    2012-01-01

    Multispecificity is not a well-understood property of some antibodies. Different functions have been attributed to multispecific natural antibodies, commonly associated with the neutralization and clearance of antigens. Much less is known about the role of antibodies like these, based on their idiotypic connectivity. B7Y33 is a chimeric IgG1 version of a polyreactive ? anti-idiotype antibody that is able to interact with different immunoglobulin and non-immunoglobulin antigens. Here we report the capacity of this antibody to enhance the immunogenicity of several autologous IgMs in adjuvant-free conditions. Our results suggest that the formation of immune complexes seems to be necessary, but not sufficient, to this activity. The potential involvement of the interaction of B7Y33 with the Fc?RIIb is discussed. PMID:22531446

  14. Molecular engineering of antibodies for therapeutic and diagnostic purposes.

    PubMed

    Ducancel, Frédéric; Muller, Bruno H

    2012-01-01

    During the past ten years, monoclonal antibodies (mAbs) have taken center stage in the field of targeted therapy and diagnosis. This increased interest in mAbs is due to their binding accuracy (affinity and specificity) together with the original molecular and structural rules that govern interactions with their cognate antigen. In addition, the effector properties of antibodies constitute a second major advantage associated with their clinical use. The development of molecular and structural engineering and more recently of in vitro evolution of antibodies has opened up new perspectives in the de novo design of antibodies more adapted to clinical and diagnostic use. Thus, efforts are regularly made by researchers to improve or modulate antibody recognition properties, to adapt their pharmacokinetics, engineer their stability, and control their immunogenicity. This review presents the latest molecular engineering results on mAbs with therapeutic and diagnostic applications. PMID:22684311

  15. Monoclonal antibody-based therapies in cancer: advances and challenges.

    PubMed

    Sapra, Puja; Shor, Boris

    2013-06-01

    Conventional anticancer therapeutics often suffer from lack of specificity, resulting in toxicities to normal healthy tissues and poor therapeutic index. Antibody-mediated delivery of anticancer drugs or toxins to tumor cells through tumor selective or overexpressed antigens is progressively being recognized as an effective strategy for increasing the therapeutic index of anticancer drugs. In this review we focus on three therapeutic modalities in the field of antibody-mediated targeting, including antibody-drug conjugates (ADCs), immunotoxins (ITs) and immunoliposomes (ILs). Design considerations for development of each of the above therapeutic modalities are discussed. Furthermore, an overview of ADCs, ITs or ILs approved for use in clinical oncology and those currently in clinical development is provided. Challenges encountered by the field of antibody-based targeting are discussed and concepts around development of the next generation of antibody therapeutics are presented. PMID:23507041

  16. Therapeutic antibodies directed at G protein-coupled receptors

    PubMed Central

    Hutchings, Catherine J; Koglin, Markus

    2010-01-01

    G protein-coupled receptors (GPCRs) are one of the most important classes of targets for small molecule drug discovery, but many current GPCRs of interest are proving intractable to small molecule discovery and may be better approached with bio-therapeutics. GPCRs are implicated in a wide variety of diseases where antibody therapeutics are currently used. These include inflammatory diseases such as rheumatoid arthritis and Crohn disease, as well as metabolic disease and cancer. Raising antibodies to GPCRs has been difficult due to problems in obtaining suitable antigen because GPCRs are often expressed at low levels in cells and are very unstable when purified. A number of new developments in overexpressing receptors, as well as formulating stable pure protein, are contributing to the growing interest in targeting GPCRs with antibodies. This review discusses the opportunities for targeting GPCRs with antibodies using these approaches and describes the therapeutic antibodies that are currently in clinical development. PMID:20864805

  17. Use of monoclonal antibodies in a radioimmunoassay for human transcortin.

    PubMed

    Faict, D; De Moor, P

    1984-03-01

    We describe the production of monoclonal antibodies to human transcortin and their use in a radioimmunoassay (RIA). A high-affinity antibody (Ka = 4 X 10(10) L/mol) made possible a sensitive RIA for transcortin (detection limit = 0.23 ng per tube), whereas use of an antibody of moderate affinity (Ka = 5 X 10(8) L/mol) was more suitable for the routine measurement of transcortin in serum, only a 25-fold dilution of the sample being required instead of 1500-fold. The correlation was good between both RIAs (r = 0.959) and between each of the RIAs and radial immunodiffusion (r = 0.955 and 0.976 for the methods with high- and low-affinity antibody, respectively). Although monoclonal antibodies were used in the RIAs and polyclonal ones in the radial immunodiffusion procedure, similar values were obtained by all techniques. PMID:6421509

  18. Characterization of a monoclonal antibody to bovine xanthine oxidase.

    PubMed Central

    Kaetzel, C S; Mather, I H; Bruder, G; Madara, P J

    1984-01-01

    The isolation of a hybridoma cell line, C-41, secreting monoclonal antibody to bovine xanthine oxidase (EC 1.2.3.2), is described. The specificity of this antibody was determined by solid-phase immunoassay, immunoblotting procedures, affinity chromatography, immunoelectrophoresis and precipitation techniques. The results are compared with those obtained in similar specificity studies on a previously described monoclonal antibody secreted by hybridoma cell line A-94 [Mather, Nace, Johnson & Goldsby (1980) Biochem. J. 188, 925-928]. This latter antibody appears to bind to xanthine oxidase only when the enzyme is immobilized on a solid support such as a plastic plate or nitrocellulose paper. Potential problems in the determination of the specificity of monoclonal antibodies, especially towards membrane proteins of unknown biological activity, are discussed. Images Fig. 2. Fig. 3. Fig. 6. Fig. 7. PMID:6378181

  19. Single-chain antibodies in pancreatic cancer.

    PubMed

    Colcher, D; Pavlinkova, G; Beresford, G; Booth, B J; Batra, S K

    1999-06-30

    Pancreatic cancer is a therapeutic challenge for surgical and medical oncology. Development of specific molecular tracers for the diagnosis and treatment of this lethal cancer has been one of our major goals. Monoclonal antibodies (MAbs) have been successfully used as selective carriers for delivering radionuclides, toxins or cytotoxic drugs to malignant cell populations; therefore, monoclonal antibody technology has led to a significant amount of research into optimizing targeted therapy. This targeted therapy results in the selective concentration of cytotoxic agents or radionuclides in tumors and should lessen the toxicity to normal tissues, which would normally limit the dosage and effectiveness of systemically administered drugs. The MAb CC49 reacts with a unique disaccharide, Sialyl-Tn, present on tumor-associated mucin (TAG-72) expressed by a majority of human adenocarcinomas. The unique Sialyl-Tn epitope has provided a potential target for immunotherapy of cancer. A single chain Fv (scFv) recombinant protein from CC49 MAb was prepared by engineering the DNA fragments for coding heavy-chain and light-chain variable regions with an appropriate oligonucleotide linker. scFv molecules, when compared to intact MAbs and the more conventional enzymatically derived F(ab')2 and Fab' fragments, offer several advantages as carriers for the selective delivery of radionuclides to tumors. The divalent antibody fragments (sc(Fv)2 or (scFv)2) display an affinity constant similar to that of the intact CC49 IgG and are stable with storage, and after radiolabeling. In preclinical studies, both the covalent and the non-covalent dimeric scFvs exhibit excellent tumor targeting properties with characteristics similar to those of the monomer, e.g., the rapid blood clearance, low kidney uptake and small size suitable for rapid penetration through tumor tissue. Increased tumor targeting of the dimers are probably due to their increased functional affinity attributable to valency, coupled with their higher molecular weight and fewer interactions with normal organs. These properties make these constructs superior to monovalent CC49 scFv. The relatively high tumor uptake, the in vitro and in vivo targeting specificity, and the stability in storage demonstrated by the dimeric CC49 sc(Fv)2 makes it a promising delivery vehicle for therapeutic applications in pancreatic cancer. PMID:10415872

  20. Antibody escape kinetics of equine infectious anemia virus infection of horses.

    PubMed

    Schwartz, Elissa J; Nanda, Seema; Mealey, Robert H

    2015-07-01

    Lentivirus escape from neutralizing antibodies (NAbs) is not well understood. In this work, we quantified antibody escape of a lentivirus, using antibody escape data from horses infected with equine infectious anemia virus. We calculated antibody blocking rates of wild-type virus, fitness costs of mutant virus, and growth rates of both viruses. These quantitative kinetic estimates of antibody escape are important for understanding lentiviral control by antibody neutralization and in developing NAb-eliciting vaccine strategies. PMID:25878104