Development of a double-antibody sandwich ELISA for rapidly quantitative detection of residual non-structural proteins in inactivated foot-and-mouth disease virus vaccines

文献类型： 外文期刊

第一作者： Fu, Yuanfang

作者： Fu, Yuanfang;Li, Dong;Cao, Yimei;Zhou, Peng;Li, Kun;Zhao, Zhixun;Li, Pinghua;Bai, Xingwen;Bao, Huifang;Liu, Zaixin;Sun, Pu;Lu, Zengjun;Wang, Songtai;Zhao, Lixia;Wang, Xiaoxiao

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关键词： Foot-and-mouth disease; Vaccine purity; Non-structural proteins; Quantitative detection; DAS-ELISA

期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：3.1； 五年影响因子：2.4 ）

ISSN： 0166-0934

年卷期： 2023 年 314 卷

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收录情况： SCI

摘要： Foot-and-mouth disease (FMD) is a highly contagious and economically devastating disease of cloven-hoofed animals. Vaccination and surveillance against non-structure protein (NSP) are the most efficacious and cost-effective strategy to control this disease. Therefore, vaccine purity control is vital for successful prevention. Currently, vaccine purity is tested by an in-vivo test that recommended in the World Organization for Animal Health (WOAH), but it is time consuming and costly. Herein, we develop a double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for quantitative detection of residual NSPs in inactivated FMD virus (FMDV) vaccines. In this assay, the monoclonal antibody 3A24 was selected as capture antibody and biotinylated 3B4B1 (Biotin-3B4B1) as detection antibody. A standard curve was developed using the NSP 3AB concentration versus OD value with the linear range of concentration of 2.5-160 ng/mL. The lowest limit of detection was 2.5 ng/mL. In addition, we determined 2.5 ng/mL of NSP as an acceptable threshold value of FMD vaccine purity using a dose-response experiment in cattle. The DAS-ELISA combined with the threshold value of FMD vaccine purity could provide a quick and simple tool for evaluation the antigenic purity of FMD vaccine during the manufacturing process.

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