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

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

doi:10.1016/j.jviromet.2023.114676

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

J Virol Methods. 2023 Apr:314:114676. doi: 10.1016/j.jviromet.2023.114676. Epub 2023 Jan 18.

Authors

Yuanfang Fu¹, Dong Li¹, Yimei Cao¹, Peng Zhou¹, Kun Li¹, Zhixun Zhao¹, Pinghua Li¹, Xingwen Bai¹, Huifang Bao¹, Songtai Wang², Lixia Zhao³, Xiaoxiao Wang⁴, Zaixin Liu¹, Pu Sun⁵, Zengjun Lu⁶

Affiliations

¹ State Key Laboratory of Veterinary Etiological Biology, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, National Foot-and-Mouth Disease Reference Laboratory, Lanzhou 730000, PR China.
² Lanzhou Animal Health Supervision, Lanzhou 730000, PR China.
³ The Spirit Jinyu Biological Pharmaceutical Co.,Ltd, Hohhot 010000 PR China.
⁴ Jingning Country Animal Disease Prevention and Control Center, Jingning 743400 PR China.
⁵ State Key Laboratory of Veterinary Etiological Biology, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, National Foot-and-Mouth Disease Reference Laboratory, Lanzhou 730000, PR China. Electronic address: sunpu@caas.cn.
⁶ State Key Laboratory of Veterinary Etiological Biology, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, National Foot-and-Mouth Disease Reference Laboratory, Lanzhou 730000, PR China. Electronic address: luzengjun@caas.cn.

PMID: 36669654
DOI: 10.1016/j.jviromet.2023.114676

Abstract

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.

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

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antibodies, Viral
Cattle
Enzyme-Linked Immunosorbent Assay / veterinary
Foot-and-Mouth Disease Virus*
Foot-and-Mouth Disease* / diagnosis
Foot-and-Mouth Disease* / prevention & control
Vaccines, Inactivated
Viral Vaccines*

Substances

Vaccines, Inactivated
Antibodies, Viral
Viral Vaccines