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Selection of chicken single-chain antibody fragments directed against recombinant VP7 of bluetongue virus
Dissertation (MSc)--University of Pretoria, 2008.
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| Format: | Thesis |
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University of Pretoria
2013
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| _version_ | 1867613704312324096 |
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| access_status_str | Open Access |
| author2 | Fehrsen, Jeanni |
| author_browse | Fehrsen, Jeanni |
| author_facet | Fehrsen, Jeanni |
| collection | Thesis |
| dc_rights_str_mv | ©University of Pretoria 2008 E1227/ |
| description | Dissertation (MSc)--University of Pretoria, 2008. |
| format | Thesis |
| id | oai:repository.up.ac.za:2263/29964 |
| institution | University of Pretoria (South Africa) |
| last_indexed | 2026-06-10T12:40:22.637Z |
| 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:2263/29964 Selection of chicken single-chain antibody fragments directed against recombinant VP7 of bluetongue virus Fehrsen, Jeanni s25522117@tuks.co.za Rakabe, Molemaisago Magdeline Chickens Bluetongue virus Viral protein seven Vp7 UCTD Dissertation (MSc)--University of Pretoria, 2008. Viral protein seven (VP7) is a major core protein and a group-reactive antigen that can be used for the diagnosis of bluetongue virus. VP7 gene of bluetongue virus serotype 4 was expressed in E. coli. Using phage display technology, anti-VP7st4 scFvs were selected from a chicken scFv library (Nkuku®) following different panning strategies. Polyclonal phage ELISA showed that VP7st4-specific scFvs were enriched after three rounds of panning. Six different scFvs (A1, H2, TA8, TC9, TD12 and SA12) were identified by sequence analysis. Stability of these scFvs was determined by incubation at different temperatures and after several freeze/thaw cycles. The scFvs were also tested in an inhibition ELISA. Inhibition with an anti-bluetongue virus guinea pig serum resulted in a 30% decrease in ELISA signal of A1. No inhibition was obtained with the rest of the scFvs when guinea pig and sheep serum were used. An anti-bluetongue virus chicken IgY inhibited the scFvs by 50% to 86%. A fragmented-gene library displaying peptides of VP7st4 was constructed. The library was subjected to three rounds of affinity selection against the anti-VP7st4 scFvs. Enrichment of clones specific to each scFv was observed. The clones were identified by sequence analyses. The regions on VP7st4 to which the scFvs bind could not be identified since no duplicate clones were selected. Veterinary Tropical Diseases unrestricted 2013-09-07T17:24:56Z 2009-04-16 2013-09-07T17:24:56Z 2008-11-28 2009-04-16 2009-02-17 Dissertation 2008 E1227/gm http://hdl.handle.net/2263/29964 http://upetd.up.ac.za/thesis/available/etd-02172009-122641/ ©University of Pretoria 2008 E1227/ application/pdf University of Pretoria |
| spellingShingle | Chickens Bluetongue virus Viral protein seven Vp7 UCTD Selection of chicken single-chain antibody fragments directed against recombinant VP7 of bluetongue virus |
| title | Selection of chicken single-chain antibody fragments directed against recombinant VP7 of bluetongue virus |
| title_full | Selection of chicken single-chain antibody fragments directed against recombinant VP7 of bluetongue virus |
| title_fullStr | Selection of chicken single-chain antibody fragments directed against recombinant VP7 of bluetongue virus |
| title_full_unstemmed | Selection of chicken single-chain antibody fragments directed against recombinant VP7 of bluetongue virus |
| title_short | Selection of chicken single-chain antibody fragments directed against recombinant VP7 of bluetongue virus |
| title_sort | selection of chicken single chain antibody fragments directed against recombinant vp7 of bluetongue virus |
| topic | Chickens Bluetongue virus Viral protein seven Vp7 UCTD |
| url | http://hdl.handle.net/2263/29964 http://upetd.up.ac.za/thesis/available/etd-02172009-122641/ |