非洲猪瘟病毒p30蛋白单克隆抗体制备及其抗原表位鉴定

doi:10.11843/j.issn.0366-6964.2023.08.026

Abstract

Abstract: This study is aimed to prepare monoclonal antibodies against African swine fever virus (ASFV) p30 protein. Five 6-8 weeks female BALB/c mice were immunized with prokaryotic recombinant p30 protein as immunogen. Spleen cells were isolated and fused with SP2/0 cells after 3 times of immunization. Two hybridoma cell lines, named 2G10-2 and 6D2-1, which could stably secrete monoclonal antibodies against ASFV p30 protein were obtained. The antibody titers were 1:12 800 and 1:3 200, respectively. The results of subtype identification showed that the heavy chain of the two monoclonal antibodies belonged to IgG, and the light chain was κ type. The results of Western blot and indirect immunofluorescence assay showed that both monoclonal antibodies had good binding activity to p30 protein. Western blot analysis showed that the two monoclonal antibodies could effectively bind the 170-194 amino acids of the recombinant p30 protein, which proved that the antigenic recognition region was the 170-194 amino acids. In this study, two monoclonal antibodies against p30 protein were prepared and their epitopes were identified, which laid a foundation for the study of the structure and function of ASFV p30 protein and the development of serological diagnostic reagents.

Key words: African swine fever virus, p30, prokaryotic expression, monoclonal antibody, epitope of antigen

CLC Number:

S852、659.1

LIU Taoxue, SU Bingqian, QI Yanli, GUO Jiangtao, LIU Zhonghu, CHU Beibei, WANG Jiang, ZENG Lei. Preparation of the Monoclonal Antibody against the African Swine Fever Virus p30 Protein and Identification of the Antigenic Epitope[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3415-3423.

References 26

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Preparation of the Monoclonal Antibody against the African Swine Fever Virus p30 Protein and Identification of the Antigenic Epitope

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