禽腺病毒血清4型Fiber2 Head蛋白表达及其单克隆抗体筛选  

Prokaryotic Expression of Fiber2 Head Protein of Fowl Adenovirus Serotype 4 and Preparation of Its Monoclonal Antibodies

作　　者：朱荣芳 金前跃[2,4,5] 柴永笑 陈晓 李伟[1,2,4,5] 郭振华[2,4,5] 卢清侠 柴书军 邢云瑞[2,4,5] 季辉 吕凤霞[7] 邢广旭 张改平[2,4,5] ZHU Rongfang;JIN Qianyue;CHAI Yongxiao;CHEN Xiao;LI Wei;GUO Zhenhua;LU Qingxia;CHAI Shujun;XING Yunrui;JI Hui;LYU Fengxia;XING Guangxu;ZHANG Gaiping(College of Life Sciences,Henan Agricultural University,Zhengzhou 450002,China;Key Laboratory of Animal Immunology,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China;College of animal medicine,Northwest A&F University,Yangling 712100,China;UK-China Centre of Excellence for Research on Avian Diseases,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou 225009,China;Jiangsu Nanjing Nongda Animal Pharmaceutical Co.,Ltd.Nanjing 210000,China;Henan Muxiang Animal Pharmaceutical Co.,Ltd,Zhengzhou 451162,China)

机构地区：[1]河南农业大学生命科学学院,郑州450002 [2]河南省农业科学院动物免疫学重点实验室,郑州450002 [3]西北农林科技大学动物医学院,杨凌712100 [4]河南省农业科学院中英禽病国际研究中心,郑州450002 [5]江苏高校动物重要疾病与人兽共患病防控协同创新中心,扬州225009 [6]江苏南京农大动物药业有限公司,南京210000 [7]河南牧翔动物药业有限公司,郑州451162

出　　处：《中国动物传染病学报》2024年第6期135-144,共10页Chinese Journal of Animal Infectious Diseases

基　　金：河南省农业科学院自主创新项目(2022ZC67);河南省农业科学院杰出青年科技基金(2024JQ01);国家重点研发计划重点专项(2016YFD0500800)。

摘　　要：为制备抗禽腺病毒血清4型(FAdV4)Fiber2 Head蛋白的特异性单克隆抗体(mAb),本研究将纤突蛋白(Fiber2)截短,并利用大肠杆菌系统进行原核表达。将纯化的蛋白免疫BALB/c小鼠,细胞融合后通过间接ELISA及免疫过氧化物酶单层细胞试验(IPMA)筛选单克隆抗体,并对单克隆抗体进行反应性测定、效价测定以及亚型鉴定。结果显示:Fiber2 Head蛋白在大肠杆菌中成功表达,且具有良好的生物活性,成功获得8株单克隆细胞株分别命名为5A5、1D5、7C9、4E5、3A11、5E11、9D7、6E3,均可分泌特异性识别Fiber2 Head蛋白及FAdV4的单克隆抗体。综上,本研究成功制备了能与Fiber2 Head蛋白和FAdV4病毒特异性反应的单克隆抗体,为FAdV4抗原和抗体检测试剂的研发奠定了基础。The purpose of this study was to prepare the specific monoclonal antibodies(mAbs)of fowl adenovirus serotype 4(FAdV4)Fiber2 Head protein.First of all,Fiber2 was truncated and expressed in prokaryotic soluble form.Then,BALB/c mice were immunized with the purified protein.After cell fusion,mAbs were screened by indirect ELISA and immunoperoxidase monolayer assay(IPMA)and determined for their reactivity,titers and subtypes.The results showed that Fiber2 Head protein was successfully expressed in prokaryotic system and had good biological activity.Eight monoclonal cell lines(5A5,1D5,7C9,4E5,3A11,5E11,9D7 and 6E3)were successfully obtained,which secreted mAbs that specifically recognized Fiber2 Head protein and FAdV4.In conclusion,mAbs that specifically reacted with Fiber2 Head protein and FAdV4 virus were successfully prepared in this study,which laid a foundation for the development of FAdV4 antigen and antibody detection reagents.

关 键 词：禽腺病毒4型 Fiber2蛋白 单克隆抗体 免疫检测 

分 类 号：S852.65[农业科学—基础兽医学]