动物医学进展2025，Vol.46Issue(11)：44-49,6.
牛冠状病毒N蛋白单克隆抗体的制备及鉴定
Preparation and Characterization of Monoclonal Antibodies against Bovine Coronavirus N Protein
摘要
Abstract
In order to prepare monoclonal antibodies against bovine coronavirus(BCoV)structural protein N,the N gene was cloned into the prokaryotic expression vector pCOLD Ⅰ,the constructed recombinant plasmid was transformed into Escherichia coli BL21(DE3),the expressed target protein was purified,and mice were immunized with purified protein to prepare hybridoma cells.After three subcloning experiments of positive hybridoma cells,ascites were prepared from mice with selected hybridoma cells,ascites reactivi-ty was detected by Western blot,and ascites reactivity was determined by indirect immunofluorescence as-say(IFA).Western blot and IFA showed the good reactivity and potency of ascites.A hybridoma cell line can stably secrete monoclonal antibody against bovine coronavirus N protein,which provides materials for further study of the function of bovine coronavirus and N protein.关键词
单克隆抗体/原核表达/牛冠状病毒/N蛋白Key words
Monoclonal antibody/Prokaryotic expression/Bovine coronavirus/N protein引用本文复制引用
闫昭宇,孔宁,张琪,许信刚,单同领,周宏超..牛冠状病毒N蛋白单克隆抗体的制备及鉴定[J].动物医学进展,2025,46(11):44-49,6.基金项目
国家重点研发计划项目(2023YFD1801300) （2023YFD1801300）
陕西省重点研发计划重点项目(2022NY-098) （2022NY-098）
规模化肉牛场疫病防控技术项目合作项目(TG20230949) （TG20230949）
