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Immunoassays with chemically modified bacteriophage

Bibliography: pages 163-176.

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Main Author: Du Plessis, Dion Henri
Format: Thesis
Language:English
Published: Department of Molecular and Cell Biology 2016
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access_status_str Open Access
author Du Plessis, Dion Henri
author_browse Du Plessis, Dion Henri
author_facet Du Plessis, Dion Henri
author_sort Du Plessis, Dion Henri
collection Thesis
description Bibliography: pages 163-176.
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institution University of Cape Town (South Africa)
language eng
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license_str Not specified — see source repository
provenance_str_mv Harvested via OAI-PMH from UCTD — University of Cape Town Open Access Repository
publishDate 2016
publishDateRange 2016
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publisher Department of Molecular and Cell Biology
publisherStr Department of Molecular and Cell Biology
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spelling oai:open.uct.ac.za:11427/17878 Immunoassays with chemically modified bacteriophage Du Plessis, Dion Henri Microbiology Bibliography: pages 163-176. The immunospecific inactivation of bacteriophage is one of the most sensitive methods available for the detection of very low concentrations of antibody. By chemically modifying the phage coat-protein, this sensitivity can be extended to antibodies against a wide variety of haptens and proteins. Phage particles that have been modified by attaching some molecule onto their surface are sensitive to antibodies directed against the coupled chemical moiety. Furthermore, the inactivation of the modified phage by antibody can be inhibited by free antigen, and this provides a sensitive assay for small quantities of antigen. Antibody and antigens have been detected at the nanogram level by this technique. The modified phage technique can also be used to distinguish antibodies of different specificities and to discriminate between closely related antigens. This technique has not yet been applied to the immunochemical study of viral components and the present work represents such an attempt. Tobacco mosaic virus (TMV) was chosen as a model system since it permits the study of numerous inununological phenomena (Rappaport, 1965; van Regenmortel, 1966). A series of preliminary experiments were performed to obtain experience in the methodology of the technique. These included the inactivation of native T4 phage by phage antiserum and anti phage IgG, the chemical modification of phage with DNP and the attachment of lysozyme to phage under a variety of conditions. The success of the covalent binding of protein to phage particles depends on finding conditions under which a proportion of the phage remains viable and, at the same time, can still be neutralized by anti-protein sera. To this end, different proportions of reactants and three different bifunctional reagents were tested. To prevent aggregation of tobacco mosaic virus protein (TMVP) at the high concentration used, the protein was treated with N-bromosuccinimide. TMVP-phage conjugates which were sensitive to antiserum were prepared using bis-diazobenzidine as the bifunctional reagent. 2016-03-17T07:12:09Z 2016-03-17T07:12:09Z 1977 Master Thesis Masters MSc http://hdl.handle.net/11427/17878 eng application/pdf Department of Molecular and Cell Biology Faculty of Science University of Cape Town
spellingShingle Microbiology
Du Plessis, Dion Henri
Immunoassays with chemically modified bacteriophage
thesis_degree_str Master's
title Immunoassays with chemically modified bacteriophage
title_full Immunoassays with chemically modified bacteriophage
title_fullStr Immunoassays with chemically modified bacteriophage
title_full_unstemmed Immunoassays with chemically modified bacteriophage
title_short Immunoassays with chemically modified bacteriophage
title_sort immunoassays with chemically modified bacteriophage
topic Microbiology
url http://hdl.handle.net/11427/17878
work_keys_str_mv AT duplessisdionhenri immunoassayswithchemicallymodifiedbacteriophage