A highly efficient indirect ELISA and monoclonal antibody established against African swine fever virus pK205R

Liwei Li; Sina Qiao; Jiachen Liu; Yanjun Zhou; Wu Tong; Shishan Dong; Changlong Liu; Yifeng Jiang; Ziqiang Guo; Haihong Zheng; Ran Zhao; Guangzhi Tong; Guoxin Li; Fei Gao

doi:10.3389/fimmu.2022.1103166

A highly efficient indirect ELISA and monoclonal antibody established against African swine fever virus pK205R

Front Immunol. 2023 Jan 9:13:1103166. doi: 10.3389/fimmu.2022.1103166. eCollection 2022.

Authors

Liwei Li¹, Sina Qiao^{1

2}, Jiachen Liu¹, Yanjun Zhou¹, Wu Tong¹, Shishan Dong², Changlong Liu¹, Yifeng Jiang¹, Ziqiang Guo¹, Haihong Zheng¹, Ran Zhao³, Guangzhi Tong¹, Guoxin Li^{1

4}, Fei Gao^{1

4}

Affiliations

¹ Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China.
² College of Veterinary Medicine, Hebei Agricultural University, Baoding, China.
³ Xiamen Center for Animal Disease Control and Prevention, Xiamen, China.
⁴ Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonose, Yangzhou University, Yangzhou, China.

Abstract

African swine fever (ASF) is a contagious infectious disease with high lethality which continuously threatens the global pig industry causing huge economic losses. Currently, there are no commercially available vaccines or antiviral drugs that can effectively control ASF. The pathogen of ASF, ASF virus (ASFV) is a double-stranded DNA virus with a genome ranging from 170 to 193 kb and 151 to 167 open reading frames in various strains, which encodes 150-200 proteins. An effective method of monitoring ASFV antibodies, and specific antibodies against ASFV to promote the development of prevention techniques are urgently needed. In the present study, pK205R of ASFV was successfully expressed in mammalian cells using a suspension culture system. An indirect enzyme-linked immunosorbent assay (ELISA) based on the purified pK205R was established and optimized. The monoclonal antibody (mAb) against pK205R recognized a conservative linear epitope (²VEPREQFFQDLLSAV¹⁶) and exhibited specific reactivity, which was conducive to the identification of the recombinant porcine reproductive and respiratory syndrome virus (PRRSV) expressing pK205R. The ELISA method efficiently detected clinical ASFV infection and revealed good application prospects in monitoring the antibody level in vivo for recombinant PRRSV live vector virus expressing the ASFV antigen protein. The determination of the conserved linear epitope of pK205R would contribute to further research on the structural biology and function of pK205R. The indirect ELISA method and mAb against ASFV pK205R revealed efficient detection and promising application prospects, making them ideal for epidemiological surveillance and vaccine research on ASF.

Keywords: 293i cells; ASFV pK205R; epitope; indirect ELISA; recombinant PRRSV.

Publication types

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

MeSH terms

African Swine Fever Virus*
African Swine Fever*
Animals
Antibodies, Monoclonal* / immunology
Enzyme-Linked Immunosorbent Assay / methods
Swine
Viral Proteins

Substances

Antibodies, Monoclonal
Viral Proteins

Grants and funding

This work was supported by the National Key Research and Development Program of China (2021YFD1801401, 2022YFD1800300, 2022YFD1800800), Shanghai Agriculture Applied Technology Development Program (2022-02-08-00-12-F01162, 2022-02-08-00-12-F01115), National Natural Science Foundation of China (31941017), and Central Public-interest Scientific Institution Basal Research Fund (Y2020YJ15, 2020JB01, 2021JB01).