Simple Summary Due to the complexity of the structure of African swine fever virus (ASFV), the structure and function of most viral encoded proteins are uncharacterized. A full understanding and identification of ASFV-coding proteins is conducive to the prevention and control of ASFV. In this study, a previously uncharacterized protein, MGF_110-13L, was identified as an immunogenic protein and expressed as a glycosylated homodimer in eukaryotic cells. Two monoclonal antibodies against MGF_110-13L were generated, and two linear epitopes were further identified using monoclonal antibodies. One epitope peptide was evaluated as a potential antigen for ASFV antibody detection. This study will provide a basis for further understanding the structure and function of the MGF_110-13L protein of ASFV. Abstract African swine fever caused by African swine fever virus (ASFV) is an acute, highly contagious swine disease with high mortality. To facilitate effective vaccine development and find more serodiagnostic targets, fully exploring the ASFV antigenic proteins is urgently needed. In this study, the MGF_110-13L was identified as an immunodominant antigen among the seven transmembrane proteins. The main outer-membrane domain of MGF_110-13L was expressed and purified. Two monoclonal antibodies (mAbs; 8C3, and 10E4) against MGF_110-13L were generated. The epitopes of two mAbs were preliminary mapped with the peptide fusion proteins after probing with mAbs by enzyme-linked immunosorbent assay (ELISA) and Western blot. And the two target epitopes were fine-mapped using further truncated peptide fusion protein strategy. Finally, the core sequences of mAbs 8C3 and 10E4 were identified as ^{48}WDCQDGICKNKITESRFIDS^{67}, and ^{122}GDHQQLSIKQ^{131}, respectively. The peptides of epitopes were synthesized and probed with ASFV antibody positive pig sera by a dot blot assay, and the results showed that epitope 10E4 was an antigenic epitope. The epitope 10E4 peptide was further evaluated as a potential antigen for detecting ASFV antibodies. To our knowledge, this is the first report of antigenic epitope information on the antigenic MGF_110-13L protein of ASFV.
【저자키워드】 monoclonal antibody, epitope, African Swine Fever Virus, MGF_110-13L protein,