Ⅰ群禽腺病毒penton蛋白单克隆抗体的制备和鉴定  

Preparation and application of monoclonal antibody against fowl adenovirus groupⅠpenton protein

作　　者：侯力丹[1,2] 陈晓春 薛麒[1] 邓永[1] 孔冬妮[1] 杨承槐[1] 刘丹[1] 李俊平[1] HOU Lidan;CHEN Xiaochun;XUE Qi;DENG Yong;KONG Dongni;YANG Chenghuai;LIU Dan;LI Junping(China Institute of Veterinary Drug Control,Beijing 100081,China;College of Veterinary Medicine,China Agricultural University,Beijing100193,China)

机构地区：[1]中国兽医药品监察所,北京100081 [2]中国农业大学动物医学院,北京100193

出　　处：《中国兽医学报》2022年第1期41-46,共6页Chinese Journal of Veterinary Science

基　　金：国家重点研发计划资助项目(2016YFD0500800,2016YFD0500809);中国兽医药品监察所公益性专项资助项目(2021012015)。

摘　　要：在同源性分析基础上,确定了12个血清型penton蛋白同源性最高的2段基因并通过linker进行连接,以该串联基因原核表达蛋白为抗原免疫小鼠,通过杂交瘤技术制备并筛选到2株可分泌FAdV-Ⅰpenton蛋白单克隆抗体的杂交瘤细胞系(Mab-penton-6#和Mab-penton-9#)。2株单克隆抗体小鼠腹水经间接免疫荧光等鉴定,可识别FAdV-Ⅰ的所有血清型代表毒株,与鸡减蛋综合征病毒、禽白血病病毒、鸡传染性喉气管炎病毒、禽呼肠孤病毒等均不发生反应,特异性良好,效价分别为1∶8000和1∶4000,最终选取Mab-penton-6#大量制备单克隆抗体。本试验成功研制出FAdV-I的penton蛋白单克隆抗体,为FAdV-Ⅰ检测方法的建立奠定了良好基础。In recent years,fowl adenovirus has been widely prevalent in China,it have caused great losses to the poultry industry.The prevalence of multiple serotypes has brought great challenges to the prevention and control of FAdV,and there is no universal monoclonal antibody for detection of FAdV-Ⅰ.In this study,homology analysis was performed on penton of FAdV-Ⅰ,and two genes with the highest homology among 12 serotypes were identified and linked by a linker.Three hybridoma cell lines secreting specific monoclonal antibodies against FAdV-Ⅰwere obtained by using prokaryotic expressed proteins as antigens.It was confirmed by indirect immunofluorescence detection that the ascites titer of the prepared monoclonal antibody was 1∶8000,and it could recognize all the representative strains of the 12 serotypes of FAdV-Ⅰ.It did not react with egg drop syndrome virus(EDSV),avian leukosis virus(ALV),avian infectious laryngeal tracheitis virus(ILTV),and avian reovirus(ARV),indicating that the prepared monoclonal antibody has good specificity.The successful development of penton monoclonal antibody for FAdV-Ⅰhas laid a good material foundation for the establishment of FAdV-Ⅰdetection method.

关 键 词：禽腺病毒I群 单克隆抗体 penton蛋白 

分 类 号：S852.4[农业科学—基础兽医学] S852.65[农业科学—兽医学]