Generation of recombinant monoclonal antibodies to study structure-function of envelope protein VP28 of white spot syndrome virus from shrimp.
Generation of recombinant monoclonal antibodies to study structure-function of envelope protein VP28 of white spot syndrome virus from shrimp.
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DOI：
10.1016/j.bbrc.2008.05.152
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发表时间：
2008-08
影响因子：
3.1
通讯作者：
Yuzhen Wang;Xiaohua Zhang;Li Yuan;T. Xu;Yu Rao;Jia Li;Heping Dai
中科院分区：
文献类型：
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作者：
Yuzhen Wang;Xiaohua Zhang;Li Yuan;T. Xu;Yu Rao;Jia Li;Heping Dai
白色斑点综合征病毒（White Spot Syndrome Virus，WSSV）是对虾养殖的主要病原。VP28是WSSV最重要的囊膜蛋白之一。本研究以大肠杆菌表达的全长VP28为抗原，免疫小鼠脾细胞，构建了噬菌体展示的单链抗体库。经过几轮淘选，从文库中分离出6个分别与VP28的N端、中部和C端区域的表位特异性结合的scFv抗体。以这些单链抗体为工具，通过免疫电镜将VP28中的表位定位在病毒体的包膜上。体外中和试验表明，这些抗原表位可能不是WSSV与宿主细胞受体的结合位点。本研究为研究WSSV囊膜蛋白的结构和功能提供了新的途径。
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.