Development of a Double-Antibody Sandwich ELISA for Rapid Detection of the MCP Antigen Concentration in Inactivated ISKNV Vaccines
文献类型： 外文期刊
作者： Liang, Hongru 1 ; Zhang, Lixi 1 ; Fu, Xiaozhe 1 ; Lin, Qiang 1 ; Liu, Lihui 1 ; Niu, Yinjie 1 ; Luo, Xia 1 ; Huang, Zhibi 1 ;
作者机构： 1.Chinese Acad Fishery Sci, Pearl River Fisheries Res Inst, Minist Agr & Rural Affairs,Key Lab Fishery Drug D, Guangdong Prov Key Lab Aquat Anim Immune Technol, Guangzhou 510380, Peoples R China
关键词： ISKNV; double-antibody sandwich ELISA; monoclonal antibody; inactivated vaccine; antigen concentration
期刊名称：VACCINES （ 影响因子：4.422； 五年影响因子：5.513 ）
ISSN：
年卷期： 2021 年 9 卷 11 期
页码：
收录情况： SCI
摘要： Infectious spleen and kidney necrosis virus (ISKNV) resulted in severe systemic diseases with high morbidity and mortality in Siniperca chuatsi. Vaccination is the primary method for effective prevention and control of these diseases. The development of inactivated ISKNV vaccines made some progress, but the technique of quality evaluation is scarce. Herein, a measurement of the MCP (major capsid protein) antigen concentration for the inactivated ISKNV vaccine was developed by double-antibody sandwich ELISA. Firstly, mouse monoclonal antibodies against ISKNV particles and MCP were generated. Then, a double-antibody sandwich ELISA was developed using the monoclonal antibody 1C8 1B9 as the capture antibody and Biotin-3B12 6B3 as the detection antibody. A standard curve was generated using the MCP concentration versus OD value with the linear range of concentration of 4.69~300 ng/mL. The assay sensitivity was 0.9 ng/mL. The antigen content of three batches of inactivated ISKNV vaccines was quantitatively detected using the double-antibody sandwich ELISA. The results showed that MCP antigen contents of inactivated ISKNV vaccines were positively correlated with the viral titers. The newly established double-antibody sandwich ELISA provided a useful tool for the detection of antigen quality for ISKNV inactivated vaccines.
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