Preparation of African swine fever virus (ASFV) p30 monoclonal antibodies and identification of novel antigenic epitopes

Abstract

African swine fever (ASF) is a highly infectious disease caused by African swine fever virus (ASFV), with a mortality rate of up to 100 % for highly virulent strains. The ASFV p30 protein is encoded by the early transcriptional gene CP204L. As one of the structural proteins of ASFV, p30 is an ideal diagnostic antigen for ASF. Here, we first generated three monoclonal antibodies (mAbs) specific for p30 from immunized BALB/c mice via cell fusion, which were successfully applied in ELISA, Western blotting, and immunofluorescence assay. Second, two novel antigenic epitopes, ¹⁶⁹TIYGTPLKE¹⁷⁷ and ¹¹¹ETNECTSSFET¹²¹ of p30 were identified using Western blotting with the three p30 mAbs. The two epitopes identified were highly conserved across genotypes I and/or II ASFVs. Third, an indirect ELISA based on epitope peptides of p30 was established to effectively detect antibodies during ASFV infection.

Keywords: African swine fever virus; Antigenic epitope; ELISA detection; Monoclonal antibodies; Structural protein p30.