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Bovine single chain Fv antibody inhibits bovine herpesvirus-1 infectivity by targeting viral glycoprotein D

文献类型： 外文期刊

作者： Xu, Jian 1 ; Wu, Jing 1 ; Jiang, Bo 1 ; He, Houjun 2 ; Zhang, Xixi 1 ; Li, Xiaoyang 1 ; Yang, Dawei 1 ; Huang, Xiufen 1 ; Se 1 ;

作者机构： 1.Beijing Acad Agr & Forestry Sci, Inst Anim Husb & Vet Med, Beijing 100097, Peoples R China

2.Jiangxi Agr Univ, Coll Anim Sci & Technol, Nanchang 330045, Jiangxi, Peoples R China

3.Beijing Agr Univ, Anim Sci & T

关键词： Bovine herpesvirus-1;Single chain Fv antibody;Glycoprotein D;Neutralizing epitope;Binding affinity;Blocking viral attachment;Virus neutralization

期刊名称：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：4.813； 五年影响因子：4.697 ）

ISSN： 0175-7598

年卷期： 2017 年 101 卷 23-24 期

页码：

收录情况： SCI

摘要： Glycoprotein D (gD) of bovine herpesvirus-1 (BoHV-1) is essential for attachment and penetration of cells during infection and is a major target for neutralizing antibodies during an adaptive immune response. Currently there are no recombinant antibodies capable of binding gD epitopes for use in treating BoHV-1 infection. In this study, a bovine scFv gene derived from a hybridoma secreting monoclonal antibodies (McAbs) against the amino acid motif MEESKGYEPP of gD was expressed in E. coli. Molecular modeling, western blot and ELISA analysis showed that this scFv had a high affinity for BoHV-1 gD, with a Kd of 161.2 +/- 37.58 nM and for whole BoHV-1 virus, with a Kd of 67.44 +/- 16.99 nM. In addition, this scFv displayed a high affinity for BoHV-1 antigen in an ELISA and competed with BoHV-1 anti-serum in a competitive ELISA. Immunofluorescence assay (IFA) and laser confocal microscopy showed that this scFv could efficiently bind to and be internalized by BoHV-1 infected Madin-Darby bovine kidney (MDBK) cells. Importantly, this scFv was shown to inhibit BoHV-1 infectivity and to reduce the number of viral plaques by blocking viral attachment to MDBK cells. Our study suggests that this bovine single-chain antibody could be developed for use as a diagnostic and therapeutic agent against BoHV-1 infection in cattle.

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