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Anchored periplasmic expression (APEx)-based bacterial display for rapid and high-throughput screening of B cell epitopes

文献类型： 外文期刊

作者： Guo, Mo 1 ; Xu, Li-Ming 2 ; Zhou, Bing 1 ; Yin, Jie-Chao 1 ; Ye, Xian-Long 1 ; Ren, Gui-Ping 1 ; Li, De-Shan 1 ;

作者机构： 1.Northeast Agr Univ, Coll Life Sci, Harbin 150030, Peoples R China

2.Chinese Acad Fishery Sci, Heilongjiang River Fishery Res Inst, Harbin 150070, Peoples R China

关键词： Anchored periplasmic expression;B cell epitotes;Epitope mapping;Fluorescence-activated cell sorting;Infectious bursal disease virus

期刊名称：BIOTECHNOLOGY LETTERS （ 影响因子：2.461； 五年影响因子：2.457 ）

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收录情况： SCI

摘要： We truncated the VP2 protein of infectious bursal disease virus into five fragments: V1-5. All fragments were displayed on the inner membrane of the Escherichia coli periplasm. After disruption of the outer membrane, spheroplasts that had anchored with the VP2 fragment were incubated with an anti-VP2 polyclonal antibody (pAb). Prey pairs were detected and quantitated by flow cytometry with V1, V3, V4 and V5 fragments reacting with the pAb. The antigenicity of all five fragments was analyzed, and our results indicated that epitopes were localized in V1, V3, V4 and V5, consistent with our flow cytometry analysis. Antigenicity analysis of purified VP2 fusion proteins using Western blots confirmed this. Our method provides a rapid, quantitative and simple strategy for identifying linear B cell epitopes.

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[1]scFv antibodies against infectious bursal disease virus isolated from a combinatorial antibody library by flow cytometry. Xu, Li-Ming,Li, Tian-He,Zhou, Bing,Guo, Mo,Liu, Miao,Zhao, Jing-Zhuang,Li, De-Shan,Cao, Hong-Wei. 2014

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