畜牧与兽医2025，Vol.57Issue(5)：99-106,8.
伪狂犬病病毒VHS蛋白单克隆抗体的制备及其抗原表位鉴定
Monoclonal antibodies preparation and antigenic epitope identification of VHS protein of pseudorabies virus
摘要
Abstract
This study was to prepare monoclonal antibody against Pseudorabies virus(PRV)VHS protein.The target gene was cloned into the prokaryotic expression vector pET-32a(+)through the prokaryotic expression system,pET-32a-VHS recombinant plasmids were ob-tained,and PRV VHS recombinant proteins were successfully prepared.The proteins were purified and inoculated into BALB/c female mice as immunogens.After cell fusion,indirect ELISA screening and 3 rounds of subcloning,two monoclonal antibodies to the PRV VHS proteins were obtained,and were named 1H8 and 4C3,respectively.The indirect ELISA assay showed that both VHS monoclonal antibody strains could secrete antibody steadily,and the titer of the ascites antibody reached 1∶409 600.The light chains of 1H8 and 4C3 belonged to the Kappa type,and the heavy chains belonged to the IgG1 subclass.Western blot and the indirect immunofluorescence assay(IFA)showed that the monoclonal antibodies secreted by the two hybridoma cell lines could react specifically with the PRV virus protein.According to the truncated expression of the VHS protein and Western blot analysis,the 1H8 monoclonal antibody could recognize epitope 241VAPEDV-KLKY250,and the 4C3 monoclonal antibody could recognize epitope 103RADGDGAADAPP114.In summary,two monoclonal antibodies to VHS were prepared and two new cell epitopes were identified here.This provided important material support for further research on the function of PRV VHS proteins and their role in viral infection and immune response.关键词
伪狂犬病病毒/VHS蛋白/单克隆抗体/抗原表位Key words
PRV/VHS protein/monoclonal antibody/antigenic epitope分类
农业科技引用本文复制引用
赵桐,潘梦娇,刘克森,白娟,马梓承,刘星..伪狂犬病病毒VHS蛋白单克隆抗体的制备及其抗原表位鉴定[J].畜牧与兽医,2025,57(5):99-106,8.基金项目
江苏省杰出青年基金项目(BK20230039,BK20241567) （BK20230039,BK20241567）
江苏省农业科技自主创新资金项目(CX(23)3073) （CX(23）
国家自然基金面上项目(32272985,32402870) （32272985,32402870）
