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

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.

Automated summary

African swine fever virus (ASFV) causes a highly contagious disease called African swine fever (ASF) in pigs, and early diagnosis is crucial for prevention and control without an effective vaccine. This study identifies P30 as an immunogenic protein produced during the early stage of ASFV infection, making it a good target for ASF detection and surveillance. By mapping the epitopes of two P30-reactive monoclonal antibodies, the study provides valuable information on the antigenic regions of ASFV P30, laying the foundation for serological diagnosis and vaccine research.