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Generation of recombinant monoclonal antibodies to study structure-function of envelope protein VP28 of white spot syndrome virus from shrimp
Authors:Wang Yuzhen  Zhang Xiaohua  Yuan Li  Xu Tao  Rao Yu  Li Jia  Dai Heping
Institution:a Institute of Hydrobiology, Chinese Academy of Sciences, 7 Southern East Lake Road, Wuchang, Wuhan, Hubei 430072, PR China
b Graduate School of the Chinese Academy of Sciences, Yuquan Road 19A, Beijing 100039, PR China
Abstract:White spot syndrome virus (WSSV) is a major pathogen in shrimp aquaculture. VP28 is one of the most important envelope proteins of WSSV. In this study, a recombinant antibody library, as single-chain fragment variable (scFv) format, displayed on phage was constructed using mRNA from spleen cells of mice immunized with full-length VP28 expressed in Escherichia coli. After several rounds of panning, six scFv antibodies specifically binding to the epitopes in the N-terminal, middle, and C-terminal regions of VP28, respectively, were isolated from the library. Using these scFv antibodies as tools, the epitopes in VP28 were located on the envelope of the virion by immuno-electron microscopy. Neutralization assay with these antibodies in vitro suggested that these epitopes may not be the attachment site of WSSV to host cell receptor. This study provides a new way to investigate the structure and function of the envelope proteins of WSSV.
Keywords:WSSV  VP28  Phage display  ScFv antibody  Shrimp
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