本研究以原核系统表达的非洲猪瘟病毒CD2v膜内区重组蛋白为免疫原接种BALB/c小鼠,经过细胞融合、间接ELISA筛选和多次亚克隆获得了8 株能够稳定分泌CD2v蛋白单克隆抗体的杂交瘤细胞系,分别命名为RH1、RH3、RH4、RH5、RH6、RH8、RH9 和RH10.8 株单克隆抗体的轻链类型均为Kappa型,其中RH1、RH5、RH9 这 3 株单克隆抗体的重链亚型为IgG2b,其余 5 株单克隆抗体的重链亚型为IgG1.Western blot和免疫荧光试验证明获得的单克隆抗体具有良好的反应性,能特异地识别昆虫杆状病毒表达系统重组表达的CD2v蛋白,可为非洲猪瘟病毒结构蛋白的功能解析以及血清学诊断方法的开发提供重要的生物学材料.
In this study,eight hybridoma cell lines which were capable of stably secreting monoclonal antibodies against CD2v protein,named RH1,RH3,RH4,RH5,RH6,RH8,RH9 and RH10,were obtained by inoculating BALB/c mice with the prokaryotic recombinant CD2v intramembrane region protein of African swine fever virus,and followed by cell fusion,indirect ELISA screening and multiple subclon-ing.The light chain type of the eight monoclonal antibodies was Kappa type.Three of the monoclonal antibodies,RH1,RH5 and RH9,were of IgG2b heavy chain subtype and the remaining five monoclonal antibodies were of IgG1 heavy chain subtype.The eight monoclonal antibod-ies showed good reactivity and specifically recognized the recombinant CD2v protein expressed by the insect baculovirus system,which might be important biological material for the functional elucidation of ASFV structural proteins and the development of serological diagnostic meth-ods.