数字农科院2.0

Identification of new antigenic epitopes of porcine reproductive and respiratory syndrome virus nsp12 protein using monoclonal antibodies

文献类型： 外文期刊

作者： Huang, Shijing;Sun, Qi;Zhu, Junrun;Wu, Xia;Liu, Ruilin;Shen, Wei;Liu, Xingyu;Yue, Ying;Gao, Fei;Jiang, Yifeng;Tong, Guangzhi;Zhou, Yanjun

作者机构：

关键词： PRRSV;nsp12 protein;Monoclonal antibody;Antigenic epitope

期刊名称： INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES

ISSN： 0141-8130

年卷期： 2024 年 283 卷

页码：

收录情况： SCIE（2024版） ; ; EI（2024版）

摘要： Porcine reproductive and respiratory syndrome virus (PRRSV), a member of the arteritis virus family, significantly impacts the swine industry due to its high infectiousness. nsp12, the nonstructural protein encoded by PRRSV, is a membrane-associated protein, with limited knowledge about its antigenic properties and functions. In this study, we used the expressed and purified nsp12 protein as antigen to immunize mice, and successfully screened five positive hybridoma cell lines that stably secrete anti-nsp12 monoclonal antibodies using immunological assays such as indirect ELISA and IFA. The antigenic epitopes recognized by the five monoclonal antibodies were identified using the fusion expression of peptides derived from the overlapping truncators of the nsp12 gene. The results showed that monoclonal antibodies 3G11 and 9C2 recognized the antigenic epitope 93 TWGFESDTAY 102 , 2E3 recognized 115 DYNDAFRARQ 124 , and 10G6 and 2A4 recognized 142 PGPVIEPTL 150 . Furthermore, the three newly identified antigenic epitopes were all immunodominant and located on the surface of nsp12 protein. Notably, the antigenic epitopes 93-102 aa are all highly conserved across PRRSV-2 strains, making them suitable targets for PRRSV-2 detection. In conclusion, our findings advance the understanding of the antigenic properties of the PRRSV nsp12 protein and facilitate the development of assays for PRRSV detection.

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