Preparation and application of porcine broadly neutralizing monoclonal antibodies in an immunoassay for efficiently detecting neutralizing antibodies against foot-and-mouth disease virus serotype O

Yimei Cao; Fengjuan Li; Xiangchuan Xing; Huiyan Zhang; QiongQiong Zhao; Pu Sun; Yuanfang Fu; Pinghua Li; Xueqing Ma; Jing Zhang; Zhixun Zhao; Hong yuan; Jian Wang; Tao Wang; Huifang Bao; Xingwen Bai; Dong Li; Qiang Zhang; Kun Li; Zengjun Lu

doi:10.1128/spectrum.02234-24

Microbiology Spectrum (Feb 2025)

Preparation and application of porcine broadly neutralizing monoclonal antibodies in an immunoassay for efficiently detecting neutralizing antibodies against foot-and-mouth disease virus serotype O

Yimei Cao,
Fengjuan Li,
Xiangchuan Xing,
Huiyan Zhang,
QiongQiong Zhao,
Pu Sun,
Yuanfang Fu,
Pinghua Li,
Xueqing Ma,
Jing Zhang,
Zhixun Zhao,
Hong yuan,
Jian Wang,
Tao Wang,
Huifang Bao,
Xingwen Bai,
Dong Li,
Qiang Zhang,
Kun Li,
Zengjun Lu

Affiliations

Yimei Cao: State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China
Fengjuan Li: State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China
Xiangchuan Xing: State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China
Huiyan Zhang: State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China
QiongQiong Zhao: State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China
Pu Sun: State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China
Yuanfang Fu: State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China
Pinghua Li: State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China
Xueqing Ma: State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China
Jing Zhang: State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China
Zhixun Zhao: State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China
Hong yuan: State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China
Jian Wang: State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China
Tao Wang: State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China
Huifang Bao: State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China
Xingwen Bai: State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China
Dong Li: State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China
Qiang Zhang: State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China
Kun Li: State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China
Zengjun Lu: State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China

DOI: https://doi.org/10.1128/spectrum.02234-24
Journal volume & issue: Vol. 13, no. 2

Abstract

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ABSTRACT Neutralizing antibodies provide vital protection against foot-and-mouth disease virus (FMDV). The virus neutralization test (VNT) is a gold standard method for the detection of neutralizing antibodies. However, its application is limited due to the requirement for live virus and unsuitability for large-scale serological surveillance. In this study, a porcine broadly neutralizing monoclonal antibody (PO18-10) against FMDV was obtained from the heterologous sequentially vaccinated pig using single-B-cell antibody technology. A competitive enzyme-linked immunosorbent assay (C-ELISA) for detecting neutralizing antibodies against FMDV serotype O was developed using biotinylated PO18-10 as a detector antibody. The sensitivity and specificity of the assay were 100% and 99.55%, respectively, and the positive/negative coincidence rate with VNT was 94%, suggesting that C-ELISA based on natural host-derived monoclonal antibody (mAb) could be a promising tool to detect neutralizing antibodies against FMDV serotype O and evaluate the vaccine efficacy.IMPORTANCEFoot-and-mouth disease virus (FMDV) serotype O is one of the most prevalent serotypes in the world. The neutralizing antibody titers in primo-vaccinated animals are directly related to their level of protection against a virus challenge. The development of a safe, rapid, and accurate method for the detection of the neutralizing antibody is essential for the control and eradication of FMD. In this study, an inter-serotype broadly neutralizing monoclonal antibody PO18-10 was successfully produced using single-B-cell antibody technology from sequentially vaccinated pigs. A competitive ELISA based on this natural host-derived mAb for the detection of neutralizing antibodies against FMDV serotype O was developed and validated. The assay demonstrates high sensitivity, specificity, and coincidence rate with VNT, making it an alternative tool for confirming FMDV infection and evaluating the vaccine efficacy.

Published in Microbiology Spectrum

ISSN: 2165-0497 (Online)
Publisher: American Society for Microbiology
Country of publisher: United States
LCC subjects: Science: Microbiology
Website: https://journals.asm.org/journal/spectrum

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