猪繁殖与呼吸综合征病毒GP3蛋白单克隆抗体制备及抗原表位鉴定

doi:10.11843/j.issn.0366-6964.2023.08.027

Abstract

Abstract: This experiment was conducted to obtain monoclonal antibodies against the GP3 protein of porcine reproductive and respiratory syndrome virus (PRRSV) and analyze its antigenic epitopes. The prokaryotic expression plasmid of PRRSV glycoprotein 3(GP3) from NADC30-like strain FJ1402 was constructed and the recombinant GP3 protein was obtained. Positive hybridoma clones were obtained through BALB/c mouse immunization and indirect ELISA. The specificity of monoclonal antibodies was further identified by indirect immunofluorescence assay and Western blot. The recognizing epitope regions of monoclonal antibodies on GP3 protein were identified by constructing truncated GP3 protein and antigenic epitope ELISA. The results showed that seven hybridoma cell lines stably secreting antibodies against the GP3 protein were successfully selected,Moreover, all monoclonal antibodies can specifically react with PRRSV FJ1402 strain. And four antigenic epitopes were identified as following,⁵⁵PLCPTRQAAAEILE⁶⁸,⁶⁹PGKSFWCRI⁷⁷, ⁷⁸GHDRCSESDH⁸⁷, and ⁸⁸DELGFMVVPPGLSS¹⁰⁰. And 2E12 mAb can also react with PRRSV BB0907 and S1 strains by using IFA and Western blot, but 4H9、9F3 and 6B3 cannot. This study successfully obtained monoclonal antibodies against GP3 protein of PRRSV, and identified four GP3 protein epitopes, laying an important foundation for the diagnosis of PRRSV and the study of the function of the viral GP3 protein.

Key words: PRRSV, GP3 protein, monoclonal antibody, epitope

CLC Number:

S852.659.6

YUAN Li, SUN Yangyang, ZHANG Lujie, ZHANG Jie, SUN Haifeng, BAI Juan, JIANG Ping. Production and Epitope Characterization of Monoclonal Antibodies against GP3 Protein of Porcine Reproductive and Respiratory Syndrome Virus[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3424-3434.

References 26

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Production and Epitope Characterization of Monoclonal Antibodies against GP3 Protein of Porcine Reproductive and Respiratory Syndrome Virus

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