Pilot-scale characterization and purification of recombinant tagged Chikungunya virus structural proteins.

Chikungunya virus (CHIKV) is swiftly emerging as a wide-reaching health menace. In this study, the sequences of recombinant tagged CHIKV structural proteins E3, E2, E1 and 6K were analysed to predict potential protein features and possible antigenic sites. All four recombinant proteins were inducibl...

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Main Author: Tan, Jiazi.
Other Authors: Richard J Sugrue
Format: Final Year Project
Language:English
Published: 2009
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Online Access:http://hdl.handle.net/10356/16355
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-163552023-02-28T18:02:51Z Pilot-scale characterization and purification of recombinant tagged Chikungunya virus structural proteins. Tan, Jiazi. Richard J Sugrue School of Biological Sciences DRNTU::Science::Biological sciences::Microbiology::Virology Chikungunya virus (CHIKV) is swiftly emerging as a wide-reaching health menace. In this study, the sequences of recombinant tagged CHIKV structural proteins E3, E2, E1 and 6K were analysed to predict potential protein features and possible antigenic sites. All four recombinant proteins were inducibly expressed in an E. coli expression system and characterised in terms of solubility in Tris-HCl/NaCl (TN) buffer, 8 M urea denaturing buffer and 6 M guanidine hydrochloride (Gn-HCl) denaturing buffer. All four recombinant proteins were found to be insoluble in TN buffer. Recombinant E3, E2 and 6K were partially soluble in 8 M urea denaturing buffer, but E1 was almost completely insoluble. 6 M Gn-HCl denaturing buffer successfully solubilised all four recombinant proteins. 6 M Gn-HCl extracts of the recombinant proteins were purified by binding to metal-affinity resins by means of the 6-histidine (6xHis) tag epitope, and elutions of the recombinant proteins obtained. Future progress will involve the scale-up of the process used here to produce purified recombinant proteins for raising monoclonal antibodies. Based on prior characterization, E1 and E2 are thus expected to be particularly suited for use as antigenic targets for raising monoclonal antibodies. Bachelor of Science in Biological Sciences 2009-05-25T07:20:38Z 2009-05-25T07:20:38Z 2009 2009 Final Year Project (FYP) http://hdl.handle.net/10356/16355 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
Tan, Jiazi.
Pilot-scale characterization and purification of recombinant tagged Chikungunya virus structural proteins.
description Chikungunya virus (CHIKV) is swiftly emerging as a wide-reaching health menace. In this study, the sequences of recombinant tagged CHIKV structural proteins E3, E2, E1 and 6K were analysed to predict potential protein features and possible antigenic sites. All four recombinant proteins were inducibly expressed in an E. coli expression system and characterised in terms of solubility in Tris-HCl/NaCl (TN) buffer, 8 M urea denaturing buffer and 6 M guanidine hydrochloride (Gn-HCl) denaturing buffer. All four recombinant proteins were found to be insoluble in TN buffer. Recombinant E3, E2 and 6K were partially soluble in 8 M urea denaturing buffer, but E1 was almost completely insoluble. 6 M Gn-HCl denaturing buffer successfully solubilised all four recombinant proteins. 6 M Gn-HCl extracts of the recombinant proteins were purified by binding to metal-affinity resins by means of the 6-histidine (6xHis) tag epitope, and elutions of the recombinant proteins obtained. Future progress will involve the scale-up of the process used here to produce purified recombinant proteins for raising monoclonal antibodies. Based on prior characterization, E1 and E2 are thus expected to be particularly suited for use as antigenic targets for raising monoclonal antibodies.
author2 Richard J Sugrue
author_facet Richard J Sugrue
Tan, Jiazi.
format Final Year Project
author Tan, Jiazi.
author_sort Tan, Jiazi.
title Pilot-scale characterization and purification of recombinant tagged Chikungunya virus structural proteins.
title_short Pilot-scale characterization and purification of recombinant tagged Chikungunya virus structural proteins.
title_full Pilot-scale characterization and purification of recombinant tagged Chikungunya virus structural proteins.
title_fullStr Pilot-scale characterization and purification of recombinant tagged Chikungunya virus structural proteins.
title_full_unstemmed Pilot-scale characterization and purification of recombinant tagged Chikungunya virus structural proteins.
title_sort pilot-scale characterization and purification of recombinant tagged chikungunya virus structural proteins.
publishDate 2009
url http://hdl.handle.net/10356/16355
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