Development of a Double-Antibody Sandwich ELISA for Rapid Detection of the MCP Antigen Concentration in Inactivated ISKNV Vaccines

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,...

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Autores principales:	Hongru Liang, Lixi Zhang, Xiaozhe Fu, Qiang Lin, Lihui Liu, Yinjie Niu, Xia Luo, Zhibin Huang, Ningqiu Li
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	ISKNV double-antibody sandwich ELISA monoclonal antibody inactivated vaccine antigen concentration Medicine R
Acceso en línea:	https://doaj.org/article/275471401ab64c2e9b1015330ace3a9d
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spelling	oai:doaj.org-article:275471401ab64c2e9b1015330ace3a9d2021-11-25T19:10:43ZDevelopment of a Double-Antibody Sandwich ELISA for Rapid Detection of the MCP Antigen Concentration in Inactivated ISKNV Vaccines10.3390/vaccines91112642076-393Xhttps://doaj.org/article/275471401ab64c2e9b1015330ace3a9d2021-11-01T00:00:00Zhttps://www.mdpi.com/2076-393X/9/11/1264https://doaj.org/toc/2076-393XInfectious 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.Hongru LiangLixi ZhangXiaozhe FuQiang LinLihui LiuYinjie NiuXia LuoZhibin HuangNingqiu LiMDPI AGarticleISKNVdouble-antibody sandwich ELISAmonoclonal antibodyinactivated vaccineantigen concentrationMedicineRENVaccines, Vol 9, Iss 1264, p 1264 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	ISKNV double-antibody sandwich ELISA monoclonal antibody inactivated vaccine antigen concentration Medicine R
spellingShingle	ISKNV double-antibody sandwich ELISA monoclonal antibody inactivated vaccine antigen concentration Medicine R Hongru Liang Lixi Zhang Xiaozhe Fu Qiang Lin Lihui Liu Yinjie Niu Xia Luo Zhibin Huang Ningqiu Li Development of a Double-Antibody Sandwich ELISA for Rapid Detection of the MCP Antigen Concentration in Inactivated ISKNV Vaccines
description	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.
format	article
author	Hongru Liang Lixi Zhang Xiaozhe Fu Qiang Lin Lihui Liu Yinjie Niu Xia Luo Zhibin Huang Ningqiu Li
author_facet	Hongru Liang Lixi Zhang Xiaozhe Fu Qiang Lin Lihui Liu Yinjie Niu Xia Luo Zhibin Huang Ningqiu Li
author_sort	Hongru Liang
title	Development of a Double-Antibody Sandwich ELISA for Rapid Detection of the MCP Antigen Concentration in Inactivated ISKNV Vaccines
title_short	Development of a Double-Antibody Sandwich ELISA for Rapid Detection of the MCP Antigen Concentration in Inactivated ISKNV Vaccines
title_full	Development of a Double-Antibody Sandwich ELISA for Rapid Detection of the MCP Antigen Concentration in Inactivated ISKNV Vaccines
title_fullStr	Development of a Double-Antibody Sandwich ELISA for Rapid Detection of the MCP Antigen Concentration in Inactivated ISKNV Vaccines
title_full_unstemmed	Development of a Double-Antibody Sandwich ELISA for Rapid Detection of the MCP Antigen Concentration in Inactivated ISKNV Vaccines
title_sort	development of a double-antibody sandwich elisa for rapid detection of the mcp antigen concentration in inactivated isknv vaccines
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/275471401ab64c2e9b1015330ace3a9d
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Development of a Double-Antibody Sandwich ELISA for Rapid Detection of the MCP Antigen Concentration in Inactivated ISKNV Vaccines

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