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Antibody phage-displayed libraries derived from chicken immunoglobulin genes : a source of highly specific diagnostic antibodies
Dissertation (MSc (Veterinary Science))--University of Pretoria, 2007.
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| Format: | Thesis |
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University of Pretoria
2013
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| _version_ | 1867613443007184896 |
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| access_status_str | Open Access |
| author2 | Du Plessis, Dion H. |
| author_browse | Du Plessis, Dion H. |
| author_facet | Du Plessis, Dion H. |
| collection | Thesis |
| dc_rights_str_mv | © 2007 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. |
| description | Dissertation (MSc (Veterinary Science))--University of Pretoria, 2007. |
| format | Thesis |
| id | oai:repository.up.ac.za:2263/25970 |
| institution | University of Pretoria (South Africa) |
| last_indexed | 2026-06-10T12:36:13.446Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from UPSpace — University of Pretoria Institutional Repository |
| publishDate | 2013 |
| publishDateRange | 2013 |
| publishDateSort | 2013 |
| publisher | University of Pretoria |
| publisherStr | University of Pretoria |
| record_format | dspace |
| source_str | UPSpace — University of Pretoria Institutional Repository |
| spelling | oai:repository.up.ac.za:2263/25970 Antibody phage-displayed libraries derived from chicken immunoglobulin genes : a source of highly specific diagnostic antibodies Du Plessis, Dion H. chilizat@hotmail.com Chiliza, Thamsanqa Emmanuel Plasmodium falciparum Monoclonal antibody (MAb) Immunoglobulin Egg yolk antibodies UCTD Dissertation (MSc (Veterinary Science))--University of Pretoria, 2007. In meeting the high demand for monoclonal antibodies, the chicken immunoglobulin system was exploited to generate recombinant antibodies against multiple target antigens. Following simultaneous immunisation of two chickens with a mixture of Plasmodium falciparum recombinant lactate dehydrogenase (LDH), histidine rich protein II (HRPII) and aldolase (ALDO), recombinant trypanosome variable surface glycoprotein (VSG) and malignant catarrhal fever virus (MCFV) each chicken produced egg yolk antibodies (IgY) against four of the five antigens. Using phage display technology, two single-chain variable fragment (scFv) antibody libraries, one with the immunoglobulin VH and VL chain regions joined by a single amino acid (G) and the other with a 15 amino acid flexible linker [(G4S) 3] were constructed using pooled splenic RNA. The single amino acid-linked scFv repertoire was evaluated as a source of highly specific diagnostic antibodies by panning against each of the five different antigens. After two rounds of panning, polyclonal phage ELISA showed the presence of antigen-specific phage antibodies against three (LDH, HRPII and VSG) of the five antigens. Five different anti-LDH and six different anti-HRPII scFvs were identified by sequence analysis. Evidence of high levels of antigen-driven gene conversion events was found in the framework and complementary determining regions and the VL chain pseudogene donors were identified. Stability of the selected scFvs was determined by incubation at different times and at different temperatures. The specificity and potential use of an LDH-specific scFv as a diagnostic reagent was shown in sandwich and competitive inhibition ELISAs. Veterinary Tropical Diseases unrestricted 2013-09-07T01:44:36Z 2008-07-16 2013-09-07T01:44:36Z 2007-11-23 2008-07-16 2008-07-01 Dissertation Chiliza, TE 2007, Antibody phage-displayed libraries derived from chicken immunoglobulin genes : a source of highly specific diagnostic antibodies, MSc dissertation, University of Pretoria, Pretoria, viewed yymmdd < http://hdl.handle.net/2263/25970 > E854/ag http://hdl.handle.net/2263/25970 http://upetd.up.ac.za/thesis/available/etd-07012008-080736/ © 2007 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. application/pdf University of Pretoria |
| spellingShingle | Plasmodium falciparum Monoclonal antibody (MAb) Immunoglobulin Egg yolk antibodies UCTD Antibody phage-displayed libraries derived from chicken immunoglobulin genes : a source of highly specific diagnostic antibodies |
| title | Antibody phage-displayed libraries derived from chicken immunoglobulin genes : a source of highly specific diagnostic antibodies |
| title_full | Antibody phage-displayed libraries derived from chicken immunoglobulin genes : a source of highly specific diagnostic antibodies |
| title_fullStr | Antibody phage-displayed libraries derived from chicken immunoglobulin genes : a source of highly specific diagnostic antibodies |
| title_full_unstemmed | Antibody phage-displayed libraries derived from chicken immunoglobulin genes : a source of highly specific diagnostic antibodies |
| title_short | Antibody phage-displayed libraries derived from chicken immunoglobulin genes : a source of highly specific diagnostic antibodies |
| title_sort | antibody phage displayed libraries derived from chicken immunoglobulin genes a source of highly specific diagnostic antibodies |
| topic | Plasmodium falciparum Monoclonal antibody (MAb) Immunoglobulin Egg yolk antibodies UCTD |
| url | http://hdl.handle.net/2263/25970 http://upetd.up.ac.za/thesis/available/etd-07012008-080736/ |