Development and characterization of monoclonal antibodies against the N-terminal domain of African swine fever virus structural protein, p54.
African swine fever virus (ASFV), a re-emerging DNA virus, causes a highly contagious disease for domestic pigs. It is running rife worldwide and threatening the global swine industry. Protein p54 is an attractive candidate for ASFV diagnostic and vaccine design. In this work, we designed a peptide to mimic the N-terminal domain (NTD) of ASFV p54 and pretested it with sera from ASFV-infected pigs. The peptide could be well recognized by the sera, implying that the NTD of p54 contained some potential linear B cell epitopes. Then, the conjugates of the peptide with bovine serum albumin were used as the immunogen to generate monoclonal antibodies (mAbs). A total of six mAbs specific to the NTD of ASFV p54 protein were developed. Five of them well reacted with ASFV HLJ/18 strain and recognized a same linear B cell epitope 5FFQPV9. Furthermore, epitope 5FFQPV9 could be well recognized by ASFV-positive sera from natural infected pigs, suggesting that it was a natural linear B-cell epitope. Conservation analysis indicated that epitope 5FFQPV9 were highly conserved among ASFV epidemic isolates belonging to genotype I and II. Alanine-scanning mutagenesis further revealed that the residues (6F to 9V) of epitope 5FFQPV9 were the core binding sites for antibody recognition. This is the first research to characterize specific mAbs against NTD of p54 protein. These findings may help further understand the function of p54 protein and the improvement of ASFV diagnosis.
['Wang|Aiping|A|', 'Jiang|Min|M|', 'Liu|Hongliang|H|', 'Liu|Yankai|Y|', 'Zhou|Jingming|J|', 'Chen|Yumei|Y|', 'Ding|Peiyang|P|', 'Wang|Yanwei|Y|', 'Pang|Weisheng|W|', 'Qi|Yanhua|Y|', 'Zhang|Gaiping|G|']
D000358:African Swine Fever Virus D000595:Amino Acid Sequence D000818:Animals D000911:Antibodies, Monoclonal D000914:Antibodies, Viral D000956:Antigens, Viral D001665:Binding Sites D018985:Epitopes, B-Lymphocyte D005838:Genotype D057809:HEK293 Cells D006801:Humans D018716:Molecular Mimicry D010455:Peptides D010802:Phylogeny D000072417:Protein Domains D012710:Serum Albumin, Bovine D013552:Swine D014162:Transfection D015678:Viral Structural Proteins
Monoclonal antibodies African swine fever virus p54 B cell epitope