Characterization of monoclonal antibody specificities against conformational epitopes of dengue NS1 glycoprotein.

Dengue is one of the most threatening mosquito-transmitted viral diseases in tropical areas. During infection, one of the disease markers, non-structural protein 1 (NS1), can be detected in the sera of infected subjects, and its serum concentration correlates with the disease severity. However, its...

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Main Author: Cheng, Wei.
Other Authors: Ruedl Christiane
Format: Final Year Project
Language:English
Published: 2009
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Online Access:http://hdl.handle.net/10356/16289
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-162892023-02-28T18:01:23Z Characterization of monoclonal antibody specificities against conformational epitopes of dengue NS1 glycoprotein. Cheng, Wei. Ruedl Christiane School of Biological Sciences DRNTU::Science::Biological sciences::Microbiology::Virology Dengue is one of the most threatening mosquito-transmitted viral diseases in tropical areas. During infection, one of the disease markers, non-structural protein 1 (NS1), can be detected in the sera of infected subjects, and its serum concentration correlates with the disease severity. However, its role in dengue pathogenesis is still unclear. The aim of this study is to characterize the specificities of 23 previously generated anti-NS1 monoclonal antibodies (mAbs) against the conformational epitopes of NS1. In this study, I used error-prone PCR to introduce mutations to NS1 gene. Then the mAbs were closely explored for their reactivity against mutant NS1 expressed on phoenix cell surface using flow cytometry. As a result, 19 mutated NS1 which caused some mAbs lose reactivity compared to wild type (wt) NS1 were selected for DNA sequencing, and the mutations were mapped around 12 critical Cysteines and on three putative domains of NS1. We believe that these identified amino acids can either be the conformational epitopes of the mAbs, or their mutations lead to conformational change in NS1, which mask the conformational epitopes of the mAbs. After characterization of their specificities, these mAbs can be explored to facilitate the crystallization of the still unknown NS1 structure. Bachelor of Science in Biological Sciences 2009-05-25T02:49:33Z 2009-05-25T02:49:33Z 2009 2009 Final Year Project (FYP) http://hdl.handle.net/10356/16289 en Nanyang Technological University 46 p. application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic DRNTU::Science::Biological sciences::Microbiology::Virology
spellingShingle DRNTU::Science::Biological sciences::Microbiology::Virology
Cheng, Wei.
Characterization of monoclonal antibody specificities against conformational epitopes of dengue NS1 glycoprotein.
description Dengue is one of the most threatening mosquito-transmitted viral diseases in tropical areas. During infection, one of the disease markers, non-structural protein 1 (NS1), can be detected in the sera of infected subjects, and its serum concentration correlates with the disease severity. However, its role in dengue pathogenesis is still unclear. The aim of this study is to characterize the specificities of 23 previously generated anti-NS1 monoclonal antibodies (mAbs) against the conformational epitopes of NS1. In this study, I used error-prone PCR to introduce mutations to NS1 gene. Then the mAbs were closely explored for their reactivity against mutant NS1 expressed on phoenix cell surface using flow cytometry. As a result, 19 mutated NS1 which caused some mAbs lose reactivity compared to wild type (wt) NS1 were selected for DNA sequencing, and the mutations were mapped around 12 critical Cysteines and on three putative domains of NS1. We believe that these identified amino acids can either be the conformational epitopes of the mAbs, or their mutations lead to conformational change in NS1, which mask the conformational epitopes of the mAbs. After characterization of their specificities, these mAbs can be explored to facilitate the crystallization of the still unknown NS1 structure.
author2 Ruedl Christiane
author_facet Ruedl Christiane
Cheng, Wei.
format Final Year Project
author Cheng, Wei.
author_sort Cheng, Wei.
title Characterization of monoclonal antibody specificities against conformational epitopes of dengue NS1 glycoprotein.
title_short Characterization of monoclonal antibody specificities against conformational epitopes of dengue NS1 glycoprotein.
title_full Characterization of monoclonal antibody specificities against conformational epitopes of dengue NS1 glycoprotein.
title_fullStr Characterization of monoclonal antibody specificities against conformational epitopes of dengue NS1 glycoprotein.
title_full_unstemmed Characterization of monoclonal antibody specificities against conformational epitopes of dengue NS1 glycoprotein.
title_sort characterization of monoclonal antibody specificities against conformational epitopes of dengue ns1 glycoprotein.
publishDate 2009
url http://hdl.handle.net/10356/16289
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