The Experts below are selected from a list of 180 Experts worldwide ranked by ideXlab platform
Anthony B Chen - One of the best experts on this subject based on the ideXlab platform.
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a sensitive Radioimmunoprecipitation assay for the detection and quantitation of antibodies to the envelope glycoprotein gp120 of the human immunodeficiency virus hiv 1
AIDS Research and Human Retroviruses, 1990Co-Authors: Catherine Lucas, Michael L Peterson, Gregory L Bennett, Steven W Frie, Phillip W Berman, Anthony B ChenAbstract:A Radioimmunoprecipitation (RIP) assay was developed to detect antibodies to the envelope glycoprotein gp120 of the human immunodeficiency virus (HIV-1). The assay, which utilized recombinant gp120 (rgp120), was quantitative, reproducible, and specific for antibodies to rgp120 or antibodies to native gp120 resulting from natural infection with HIV. Polyethylene glycol-8000 (PEG), used in the assay at a final concentration of 10% to precipitate immune complexes, was demonstrated to be effective in titering sera from different animal species. Provided samples were diluted at least 1:100, antibody titers could be determined either by the classical dilution method or by interpolation from a calibration curve prepared with a positive serum. The humoral response of animals immunized with rgp120 was monitored and a positive correlation was found between titers determined in the RIP assay and the ability of the sera to neutralize. In addition, RIP titers of HIV-positive human sera correlated very well with reactivity obtained in a commercial HIV immunoblot assay. The assay has the advantage of quantitation, fast turnaround time, and versatility.
George J Dawson - One of the best experts on this subject based on the ideXlab platform.
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discrimination of hiv 2 infection from hiv 1 infection by western blot and Radioimmunoprecipitation analysis
AIDS Research and Human Retroviruses, 1990Co-Authors: Timothy J Holzer, Rhonda G Allen, Cynthia A Heynen, Mark M Kennedy, Mark F Knigge, Deborah A Paul, George J DawsonAbstract:Serum and plasma samples were collected from blood donors who were confirmed positive for antibodies to HIV-1 in the United States, and from blood donors and individuals in West Africa and Portugal who were positive for antibodies to HIV-1, HIV-2, or both. Western blots and sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) Radioimmunoprecipitation assays (RIPA) utilizing native HIV-1 and HIV-2 proteins were performed on these specimens to determine the ability of these procedures to discriminate between HIV-1 and HIV-2 infections. Extensive serologic cross reactivity between HIV-1 and HIV-2 p24 was found in both populations. Antibody reactivity to the envelope protein gp120 was able to discriminate 20 of 20 (100%) U.S. specimens as HIV-1 infections. In specimens from West Africa and Portugal, Western blot and RIPA were in complete agreement on 33 of 42 samples (78.6%). Among these 33 specimens, 10 were found to be reactive for antibodies to HIV-1 only, 10 were reactive to HIV-2 only, and...
Angela Vincent - One of the best experts on this subject based on the ideXlab platform.
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a sensitive Radioimmunoprecipitation assay for assessing the clinical relevance of antibodies to ifn beta
Journal of Neurology Neurosurgery and Psychiatry, 2003Co-Authors: N Lawrence, J Palace, Joel Oger, T Aziz, Angela VincentAbstract:Background: Some multiple sclerosis (MS) patients treated with interferon beta (IFN s) develop antibodies to the drug. Neutralising antibody (NAB) assays for IFN s are expensive and the clinical relevance of the results has been debated. Objective: To establish a cheap, sensitive, and reliable assay for antibodies to 125I-IFN s, and to correlate levels of antibodies with clinical response to IFN s treatment. Methods: We established a Radioimmunoprecipitation assay (RIPA) using 125I-IFN s. We tested NAB positive sera, healthy control sera, and serial samples of 33 IFN s-1b treated MS patients from the Vancouver cohort of the Berlex pivotal trial who had a high incidence of NABs. Results: We found that the RIPA was highly sensitive for the detection of antibodies to IFN s-1a and -1b, and that there was a strong correlation between reactivity of NAB positive sera for 125I-IFN s-1b and for 125I-IFN s-1a. The RIPA was more sensitive and consistent than the NAB. Moreover, there was a trend towards poorer MRI outcomes in RIPA positive patients, but not in NAB-positive patients. Conclusions: The RIPA assay is sensitive and easy to perform. It should be of value in assessing the clinical impact of IFN s antibodies, and its use could help target expensive INF s treatments to those who will respond best.
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a Radioimmunoprecipitation assay for antibodies to botulinum a
Neurology, 1998Co-Authors: J Palace, Angela Nairne, N Hyman, T V Doherty, Angela VincentAbstract:We quantified antibodies to botulinum A (anti-BTx) by immunoprecipitation of 125I-BTx. We tested seven bioassay-positive sera and 68 coded samples, including 18 from patients who had ceased to respond to BTx treatment. Compared with values from healthy control subjects and 42 neurologic control subjects, all bioassay-positive sera were positive (range, 258 to 2,809 pM) and 49 of 50 patients who continued to respond to BTx were negative (
D J Jackwood - One of the best experts on this subject based on the ideXlab platform.
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comparison of neutralizing epitopes among infectious bursal disease viruses using Radioimmunoprecipitation
Avian Diseases, 1995Co-Authors: P L Whetzel, D J JackwoodAbstract:SUMMARY. Monoclonal antibodies (MAbs) were used to analyze the relatedness of neutralizing epitopes of infectious bursal disease virus (IBDV) strains. The MAb B69 neutralized the homologous D78 virus but not the MD and Del-A variant strains of IBDV. MAbs 33E8 and R63 neutralized D78 and variant strains MD and Del-A. A cDNA clone consisting of a 1000base-pair fragment of the VP2 gene from the Del-A IBDV strain was translated using an in vitro system. Six peptides were observed following translation, which represented a fulllength product (35.5 kilodaltons) and five truncated products. The translated peptides were radioimmunoprecipitated using MAbs B69, 33E8, and R63. Only MAb R63 immunoprecipitated the in vitro translation products from Del-A. MAbs B69 and 33E8 did not immunoprecipitate the in vitro-translated VP2 peptides. The D78 and Del-A viruses were radiolabeled in vivo using 35S-methionine. Proteins from these viruses were examined by Radioimmunoprecipitation with MAbs B69, 33E8, and R63. Although the background made interpretation of the results difficult for MAb B69, this MAb clearly immunoprecipitated VP2 of D78. MAb 33E8 immunoprecipitated the D78 VP3 protein, and R63 immunoprecipitated the VP2 protein. MAb R63 also immunoprecipitated the 35S-methionine-labeled VP2 protein from Del-A. It was concluded that the neutralizing epitope represented by 33E8 was located on VP3 and that the epitope represented by R63 is located on both classic and variant viruses in the region of VP2 that is generally thought to contain sequence variability among IBDV strains.
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evaluation of vp2 epitopes of infectious bursal disease virus using in vitro expression and Radioimmunoprecipitation
Archives of Virology, 1993Co-Authors: J M Crisman, R J Jackwood, D P Lana, D J JackwoodAbstract:A clone (pV 17-7) spanning a portion of the VP2 gene of infectious bursal disease virus (IBDV) was selected from a cDNA library produced using the variant A virus strain. This clone was expressed in vitro and the protein products were immunoprecipitated with various virus-neutralizing antisera made against 6 different strains of IBDV. The antisera made against 4 variant strains immunoprecipitated the translation products from the pV 17-7 clone, but the antisera to the classic STC virus and the serotype 2 OH virus did not immunoprecipitate the pV 17-7 translation products.
Raphael P Viscidi - One of the best experts on this subject based on the ideXlab platform.
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detection of antibodies to the major outer membrane protein of chlamydia trachomatis using an in vitro transcription translation Radioimmunoprecipitation assay
Serodiagnosis and Immunotherapy in Infectious Disease, 1996Co-Authors: Janice R Verley, Judith A Whittumhudson, Thomas C Quinn, Raphael P ViscidiAbstract:Abstract A Radioimmunoprecipitation assay (RIPA) using in vitro translated protein was developed to measure serum antibodies to the major outer membrane protein (MOMP) of C. trachomatis . Polyclonal animal antisera to C. trachomatis serovars C, E and F reacted to serogroup-specific determinants by RIPA, as compared to species-specific epitopes by Western blot. Antibody responses in monkeys immunized systemically or mucosally with a candidate MOMP vaccine were assessed by RIPA and ELISA with elementary bodies (EB-ELISA). Unlike EB-ELISA, RIPA showed a significant difference in pre-challenge antibody levels between systemically and mucosally immunized animals. Additionally, only RIPA showed a significant inverse correlation between antibody level at time of challenge and microbiologic response measured as median inclusion forming units (IFU) in culture ( r = −0.89; P C. trachomatis antigens and may be more informative than EB-ELISA and Western blot for assessing the immunogenicity of MOMP vaccines.
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comparison of peptide enzyme linked immunosorbent assay and Radioimmunoprecipitation assay with in vitro translated proteins for detection of serum antibodies to human papillomavirus type 16 e6 and e7 proteins
Journal of Clinical Microbiology, 1994Co-Authors: Keerti V Shah, M Muller, Nubia Munoz, X F Bosch, Raphael P ViscidiAbstract:Antibodies to human papilloma virus (HPV) type 16 (HPV-16) E6 and E7 proteins in serum are markers for HPV-associated invasive cervical carcinoma. We compared two assays, a Radioimmunoprecipitation assay with in vitro-translated HPV-16 E6 and E7 proteins and an enzyme-linked immunosorbent assay (ELISA) with E6 and E7 synthetic peptides, for their abilities to discriminate serologically between patients with invasive cervical cancer and controls. Among the patients, antibody prevalences were higher by the E6 Radioimmunoprecipitation assay (55.7%) than by the E6 peptide ELISA (15.5%), but among the controls, they were lower by the Radioimmunoprecipitation assay (1.7%) than by the E6 peptide ELISA (5%). For E7, antibody prevalences among the patients were comparable by the Radioimmunoprecipitation assay (43%) and the peptide ELISA (41%), but among the controls they were higher by the E7 peptide ELISA (17.4%) than by the Radioimmunoprecipitation assay (4.1%). There was good agreement between the E7 Radioimmunoprecipitation assay and the E7 peptide ELISA among patients but not among controls. In tests with representative sera, heat denaturation of the translated proteins resulted in a complete loss of reactivity to the E6 protein and a marked decrease in reactivity to the E7 protein. Our study showed that the Radioimmunoprecipitation assay discriminates better than the peptide ELISA between patients with invasive cervical cancer and controls and that this is related to the ability of the Radioimmunoprecipitation assay to detect conformational epitopes. Images
