Development Of An Immune Tuberculosis Antibody Library: Monoclonal Antibody Generation For a-Crystalline Antigen
| dc.contributor.author | Hamidon, Nurul Hamizah | |
| dc.date.accessioned | 2019-08-29T07:08:11Z | |
| dc.date.available | 2019-08-29T07:08:11Z | |
| dc.date.issued | 2017-05 | |
| dc.description.abstract | Antibodies are very important for therapeutic and diagnostic applications for infectious diseases. Immune libraries are useful for infectious disease applications because it would allow for better and selective enrichment of antibodies against disease antigens. The generations of antibody libraries are necessary to produce the monoclonal antibodies. Immune scFv antibody library was generated from six donors (tuberculosis patients). Conventional PCR method was used to amplify all possible antibody genes, which were then cloned into the pLABEL, phagemid vector. A two-step cloning procedure was done to generate the antibody library. The library size of the mini HC library and the scFv library constructed was approximately around 107 and 109 individual clones respectively. The insert rate for two-step cloning was only 63% and -crystalline antigen was used for verification of the antibody library and monoclonal isolation. Conventional plate panning technique was used for selection of monoclonal antibodies. Three monoclonal scFv antibodies were successfully obtained for the Mycobacterium tuberculosis, -crystalline (Rv2331c) antigen and the antibody sequence was analyzed using the software, VBASE2. The clones were IGKV3 and IGHV2 for clone 1H, IGKV4 and IGHV2 for clone 1B and IGLV1 and LGHV1 for clone 10G. The antibodies enriched was also evaluated in soluble form for binding activities. In conclusion, a diverse immune scFv library was constructed from which soluble monoclonal scFv antibodies against a-crystalline antigen was obtained. | en_US |
| dc.identifier.uri | http://hdl.handle.net/123456789/8734 | |
| dc.language.iso | en | en_US |
| dc.publisher | Universiti Sains Malaysia | en_US |
| dc.subject | Development of an immune tuberculosis antibody | en_US |
| dc.subject | monoclonal antibody generation | en_US |
| dc.title | Development Of An Immune Tuberculosis Antibody Library: Monoclonal Antibody Generation For a-Crystalline Antigen | en_US |
| dc.type | Thesis | en_US |
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