您好，欢迎访问中国农业科学院 机构知识库！

全部

  • 全部
  • 标题
  • 学者
  • 机构
  • 关键词

Single-B-cell cloning and recombinant antibodies generation to analyze the antigenicity of porcine reproductive and respiratory syndrome virus nonstructural protein 12

文献类型： 外文期刊

作者： Duan, Hongyong 1 ; Tang, Chaozhi 1 ; Han, Song 2 ; Yang, Nan 2 ; Wang, Shumao 2 ; Gao, Fei 2 ; Zhou, Yanjun 2 ; Tong, Guangzhi 2 ; Zhao, Kuan 3 ; Li, Liwei 2 ;

作者机构： 1.Henan Normal Univ, Coll Life Sci, Xinxiang 453007, Peoples R China

2.Chinese Acad Agr Sci, Shanghai Vet Res Inst, 518 Ziyue Rd, Shanghai 200241, Peoples R China

3.Hebei Agr Univ, Coll Vet Med, Baoding 071001, Peoples R China

4.Yangzhou Univ, Jiangsu Coinnovat Ctr Prevent & Control Important, Yangzhou 225009, Peoples R China

关键词： PRRSV; Nsp12; Antigenic epitope; Recombinant mAbs; Single-B-cell antibody technology

期刊名称：VETERINARY MICROBIOLOGY （ 影响因子：2.7； 五年影响因子：2.9 ）

ISSN： 0378-1135

年卷期： 2025 年 304 卷

页码：

收录情况： SCI

摘要： The prevalence and variation of porcine reproductive and respiratory syndrome virus (PRRSV) in China are increasing. The rapid preparation of essential antibodies will effectively reveal the antigenicity, epitopes, and intracellular distribution of viral proteins. Single-B-cell antibody technology is a novel method for screening diverse functional monoclonal antibodies (mAbs). Herein, we successfully expressed PRRSV nonstructural protein 12 (Nsp12) in suspension-cultured Chinese hamster ovary (CHO) cells. Using single-B-cell antibody technology, we utilized fluorescence-activated cell sorting to collect individual immune B cells and prepared singlecell reverse transcription-polymerase chain reaction to clone the variable region of immunoglobulin heavy chain (IgH) and immunoglobulin light chain (IgK). Two recombinant mAbs were generated via transient transfection of CHO cells with the corresponding expression plasmids of IgH and IgK. A novel linear epitope (104YEFTGNGEDW113) of Nsp12 was identified using mAb1N14. This epitope was conserved in lineages 1, 5, and 8 of PRRSV-2 and was located on the surface of the Nsp12 spatial structure. The amino acid mutation in Nsp12 of lineage 3 PRRSV-2 affected the antigenicity of this linear epitope. A conserved conformational epitope was identified using mAb2S18, and the spatial structure of Nsp12 showed high similarity between PRRSV-1 and different lineages of PRRSV-2. During PRRSV infection, Nsp12 was distributed in the cytoplasm and accumulated in the nucleus. Overall, antigenicity analysis and novel epitope identification contributed to the in-depth exploration of the biological function of Nsp12 and will facilitate the development of detection assays and antiviral strategies.

  • 相关文献
作者其他论文 更多>>
置顶
购物车
反馈

© 京ICP备09089781号-29 主办单位：中国农业科学院

学术资源： 中国农业科学院 农业专业知识服务系统 农科联盟资源共建共享平台 中国农业科技文献与信息服务平台 中国农业期刊集成服务平台