Preparation and epitope mapping of monoclonal antibodies against African swine fever virus P30 protein

• Published in: Applied microbiology and biotechnology 2022-02, Vol.106 (3), p.1199-1210
• Main Authors: Zhou, Gaijing, Shi, Zhengwang, Luo, Juncong, Cao, Liyan, Yang, Bo, Wan, Ying, Wang, Lijuan, Song, Rui, Ma, Yuan, Tian, Hong, Zheng, Haixue
• Format: Article
• Language: English
• Subjects: African Swine Fever; African Swine Fever Virus - immunology; Alanine; Amino acids; Animals; Antibodies, Monoclonal - immunology; Antibodies, Viral; Antigenic determinants; Antigens; Applied Genetics and Molecular Biotechnology; Asfarviridae; Biomedical and Life Sciences; Biotechnology; Diagnosis; Epitope Mapping; Epitopes - genetics; Fever; Genetic aspects; Genotypes; Identification and classification; Immunization; Immunogenicity; Life Sciences; Mice; Microbial Genetics and Genomics; Microbiology; Monoclonal antibodies; p30 Protein; Peptide mapping; Peptides; Polypeptides; Proteins; Serology; Swine; Synthetic peptides; Target detection; Vaccines; Viral Proteins - immunology; Viruses
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-022-11784-7

African swine fever virus (ASFV) causes acute, febrile, and highly contagious diseases in swine. Early diagnosis is critically important for African swine fever (ASF) prevention and control in the absence of an effective vaccine. P30 is one of the most immunogenic proteins that are produced during the early stage of an ASFV infection. This makes P30 a good serological target for ASF detection and surveillance. In this study, two P30-reactive monoclonal antibodies (mAbs), 2H2 and 5E8, were generated from mice immunized with recombinant P30 protein (rP30). Epitope mapping was performed with overlapping polypeptides, alanine mutants, and synthetic peptides. The mapping results revealed that 2H2 recognized a region located in the N-terminal, 16–48 aa. In contrast, 5E8 recognized a linear epitope in the C-terminal, 122–128 aa. Further analysis indicated that the epitope recognized by 2H2 was highly conserved in genotypes I and II, while the 5E8 epitope was conserved in most genotypes and the Ser to Pro change at position 128 in genotypes IV, V, and VI did not affect recognition. Overall, the results of this study provide valuable information on the antigenic regions of ASFV P30 and lay the foundation for the serological diagnosis of ASF and vaccine research. Key points • Two specific and reactive mAbs were prepared and their epitopes were identified. • 2H2 recognized a novel epitope highly conserved in genotypes I and II. • 5E8 recognized a seven-amino acid linear epitope highly conserved in most genotypes.