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
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 unsuitabili...
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American Society for Microbiology
2025-02-01
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| Series: | Microbiology Spectrum |
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| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.02234-24 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Yimei Cao |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-3d32544b91784e39a59976a184f18026 |
| institution | Kabale University |
| issn | 2165-0497 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj-art-3d32544b91784e39a59976a184f180262025-02-04T14:03:41ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972025-02-0113210.1128/spectrum.02234-24Preparation and application of porcine broadly neutralizing monoclonal antibodies in an immunoassay for efficiently detecting neutralizing antibodies against foot-and-mouth disease virus serotype OYimei Cao0Fengjuan Li1Xiangchuan Xing2Huiyan Zhang3QiongQiong Zhao4Pu Sun5Yuanfang Fu6Pinghua Li7Xueqing Ma8Jing Zhang9Zhixun Zhao10Hong yuan11Jian Wang12Tao Wang13Huifang Bao14Xingwen Bai15Dong Li16Qiang Zhang17Kun Li18Zengjun Lu19State 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, ChinaState 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, ChinaState 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, ChinaState 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, ChinaState 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, ChinaState 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, ChinaState 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, ChinaState 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, ChinaState 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, ChinaState 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, ChinaState 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, ChinaState 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, ChinaState 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, ChinaState 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, ChinaState 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, ChinaState 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, ChinaState 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, ChinaState 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, ChinaState 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, ChinaState 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, ChinaABSTRACT 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.https://journals.asm.org/doi/10.1128/spectrum.02234-24foot-and-mouth disease virusporcine broadly neutralizing monoclonal antibodycompetitive ELISAneutralizing antibodyserotype O |
| spellingShingle | 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 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 Microbiology Spectrum foot-and-mouth disease virus porcine broadly neutralizing monoclonal antibody competitive ELISA neutralizing antibody serotype O |
| title | 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 |
| title_full | 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 |
| title_fullStr | 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 |
| title_full_unstemmed | 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 |
| title_short | 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 |
| title_sort | 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 |
| topic | foot-and-mouth disease virus porcine broadly neutralizing monoclonal antibody competitive ELISA neutralizing antibody serotype O |
| url | https://journals.asm.org/doi/10.1128/spectrum.02234-24 |
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