High throughput screening of recombinant antibodies against infectious hematopoietic necrosis virus from a combinatorial antibody library

文献类型： 外文期刊

第一作者： Xu, Li-Ming

作者： Xu, Li-Ming;Zhao, Jing-Zhuang;Liu, Miao;Cao, Yong-Sheng;Yin, Jia-Sheng;Liu, Hong-Bai;Tongyan-Lu

作者机构：

关键词： Antibody library;Antibody-antigen co-expression;Bacterial display;Glycoprotein;Infectious hematopoietic necrosis virus

期刊名称：AQUACULTURE （ 影响因子：4.242； 五年影响因子：4.723 ）

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收录情况： SCI

摘要： Infectious hematopoietic necrosis virus (IHNV) is a significant rhabdoviral pathogen of salmonid fish. In this study, a single chain variable fragment (scFv) antibody library derived from rainbow trout (Oncorhynchus mykiss) and a glycoprotein fragment (named G4, 540 nt, 180 aa) of IHNV-Sn1203 isolate were co-expressed by a bacterial display technology. The library was subjected to three rounds of screening by flow cytometry (FCM) to select IHNV specific antibodies. Seven antibody clones with different mean fluorescence intensities (MFI) were obtained by picking colonies at random. The antibody clone with the highest MFI was expressed and purified. The purified IHNV-specific scFv antibody was used successfully in Western blotting, enzyme linked immunosorbent assay and an immunofluorescence antibody test. This method provides a high throughput means to screen an antibody library by flow cytometry and isolate an antibody that can be used as a potential universal reagent for the detection and confirmation IHNV strains that are prevalent throughout China.

分类号： S9

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