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Development of GEM-PA-nanotrap for purification of foot-and-mouth disease virus

文献类型： 外文期刊

作者： Cheng, Haiwei 1 ; Chen, Jin 1 ; Cai, Zizheng 4 ; Du, Luping 1 ; Hou, Jibo 1 ; Qiao, Xuwen 1 ; Zheng, Qisheng 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Inst Vet Immunol & Engn, Nanjing 210014, Jiangsu, Peoples R China

2.Jiangsu Acad Agr Sci, Key Lab Vet Biol Engn & Technol, Minist Agr, Natl Res Ctr Engn & Technol Vet Biol, Nanjing 210014, Jiangsu, Peoples R China

3.Jiangsu Coinnovat Ctr Prevent & Control Important, Yangzhou 225009, Jiangsu, Peoples R China

4.Nanjing Agr Univ, Nanjing 210095, Jiangsu, Peoples R China

关键词： FMDV; Affinity purification; Gram-positive enhancer matrix particles; Nanobody; Nanotrap

期刊名称：VACCINE （ 影响因子：3.641； 五年影响因子：3.816 ）

ISSN： 0264-410X

年卷期： 2019 年 37 卷 24 期

页码：

收录情况： SCI

摘要： Vaccination is the primary preventative measure against outbreaks of foot-and-mouth disease (FMD). The efficacy of inactivated FMD vaccines is mainly determined by the integrity of foot-and-mouth disease virus (FMDV) particles (referred to as 146S particles), and impurities in the inactivated vaccines could result in side effects. In this study, we developed an effective affinity purification method for the purification of FMDV from cellular lysates, referred to as GEM-PA-nanotrap. To develop the GEM-PA-nanotrap, a nanobody (Nb205) against FMDV vaccine strain O/MYA98/BY/2010 146S particles was selected from a non-immunized library and fused to a peptidoglycan-binding protein anchor (PA). The PA-Nb205 fusion protein was non-covalently coupled to the surface of Gram-positive enhancer matrix (GEM) particles, which were prepared from the non-living, non-genetically modified, Gram-positive, food-grade Lactococcus lactis bacteria. The GEM-PA-nanotrap was used to purify FMDV from cellular lysates through a simple incubation and centrifugation step. The FMDV recovery rate was more than 99%, the efficiency of nonviral protein removal was about 98.3%, and the purification process had almost no effect on the integrity and immunogenicity of 146S particles. Therefore, the GEM-PA-nanotrap has potential as an effective method for the recovery and purification of FMDV during the vaccine manufacturing process. (C) 2019 Elsevier Ltd. All rights reserved.

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