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Title: Identification and molecular characterization of human antibody fragments specific for dengue NS5 protein
Authors: Zhao, Y.
Moreland, N.J.
Tay, M.Y.F.
Lee, C.C.
Swaminathan, K. 
Vasudevan, S.G. 
Keywords: Antibody
Phage display
Issue Date: 22-Jan-2014
Citation: Zhao, Y., Moreland, N.J., Tay, M.Y.F., Lee, C.C., Swaminathan, K., Vasudevan, S.G. (2014-01-22). Identification and molecular characterization of human antibody fragments specific for dengue NS5 protein. Virus Research 179 (1) : 225-230. ScholarBank@NUS Repository.
Abstract: The multifunctional dengue nonstructural (NS) protein 5 from the four serotypes of dengue virus (DENV1-4) is essential for viral replication and harbors a methyl transferase (MTase) and a RNA-dependent RNA-polymerase domain (RdRp). There are limited comparative studies of NS5 from the four DENV serotypes and this is further hampered by a lack of cross-reactive NS5 antibodies. In this study, recombinant NS5 proteins were expressed, purified, enzymatically characterized, and used strategically as bait in biopanning experiments with a naïve human Fab phage-display library to identify serotype specific or cross-reactive Fab fragments. Using a combination of peptide competition ELISA and peptide phage display the epitopes of the cross-reactive Fabs were mapped to the first alpha helix of the MTase domain (5M1) and the priming loop of the RdRp domain (5R3). The epitope of a third, serotype-specific Fab (5M3) was mapped to aa19-30 of the DENV3 MTase domain. Together the recombinant proteins and specific antibodies will facilitate further mechanistic studies of the DENV replication complex. © 2013 Elsevier B.V.
Source Title: Virus Research
ISSN: 01681702
DOI: 10.1016/j.virusres.2013.11.010
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