The Novel Antigenic Epitopes of ASFV Inner Membrane p54 Protein Revealed by Monoclonal Antibodies

African swine fever (ASF) is caused by the African swine fever virus (ASFV), infection in domestic pigs and wild boars causes a highly contagious, hemorrhagic disease. The p54 protein is encoded by the ASFV E183L gene and is one of the important structural proteins located on the inner envelope of the virus. It is involved in processes of virus assembly, apoptosis induction and neutralizing antibody production. In this study, three specific monoclonal antibodies (mAbs) against ASFV p54 protein were generated, namely 6B11, 3E3, and 3C10, from the mice immunized with recombinant prokaryotic p54 truncated protein. Three novel linear B cell epitopes recognized by the mAbs were revealed, which are 60AAIEEEDIQFINP72, 128MATGGPAAAPAAASAPAHPAE148, and 163MSAIENLRQRNTY175. The epitopes 60AAIEEEDIQFINP72 and 163MSAIENLRQRNTY175 were highly conserved in genotype I and II ASFV strains. In addition, the epitope peptide ELISA can be used for the detection of ASFV antibody. Our work provides new insights for vaccine development and tools for serological diagnosis of ASF.