猪圆环病毒2型Rep蛋白单克隆抗体的制备及鉴定  

Preparation and Identification of Monoclonal Antibodies against Rep Protein of Porcine Circovirus Type 2

作　　者：高梅[1] 彭喆[2] 吴晓燕[2] 时建立[1,2] 辛长勋 郑书轩 刘玉伟 王硕[1] 王金宝[1,2] 李俊[1,2] GAO Mei;PENG Zhe;WU Xiaoyan;SHI Jianli;XIN Changxun;ZHENG Shuxuan;LIU Yuwei;WANG Shuo;WANG Jinbao;LI Jun(College of Life Sciences,Shandong Normal University,Jinan,Shandong 250014,China;Institute of Animal Science and Veterinary Medicine,Shandong Academy of Agricultural Sciences,Jinan,Shandong 250101,China;Veterinary Medicine Department,Qingdao Agricultural University,Qingdao,Shandong 266109,China)

机构地区：[1]山东师范大学生命科学院,山东济南250014 [2]山东省农业科学院畜牧兽医研究所,山东济南250100 [3]青岛农业大学动物医学院,山东青岛266109

出　　处：《家畜生态学报》2018年第10期59-63,共5页Journal of Domestic Animal Ecology

基　　金：山东省农业科学院青年科研基金项目(2016YQN53);"十三五"国家重点研发计划子课题(2016YFD0500708);山东省现代农业产业技术体系疫病控制岗位(SDAIT-08-07);山东省农业重大应用创新项目;泰山学者项目

摘　　要：本研究旨在制备猪圆环病毒2型(PCV2)Rep蛋白单克隆抗体。PQE30-PCV2Rep蛋白在IPTG的诱导下表达,用SDS-PAGE对表达产物进行检测。将纯化的重组蛋白免疫小鼠,制备单克隆抗体,ELASA法鉴定抗体亚型及效价。结果显示重组蛋白PQE30-PCV2Rep得到高效表达,筛选获得了3株稳定分泌抗Rep蛋白单克隆抗体的杂交瘤细胞系1G4、2G4和6H12,其分泌的抗体分别为IgG2a、IgG2b、IgG2b亚型。其中,6H12抗体效价为256K,浓度为0.47mg/mL,纯度>90%。本研究制备的单克隆抗体有较好的生物学特性,为PCV2的研究及快速诊断提供了新的方法。The objective of this study is to prepare monoclonal antibodies against Rep protein of porcine circovirus type 2(PCV2).The PQE30-PCV2 Rep protein was expressed under IPTG induction,and the expression products were identified by SDS-PAGE.The purified recombinant protein was used to immunize mice in order to prepare monoclonal antibodies,and ELASA method was used to identify antibody subtypes and titers.The results showed that the recombinant protein PQE30-PCV2 Rep was highly expressed,and 3 hybridoma cell lines 1G4,2G4 and 6H12 monoclonal antibodies against Rep protein were screened out,their antibody subtypes were IgG2a,IgG2b and IgG2b,respectively.The titer of 6H12 antibody was 256K,the concentration was 0.47 mg/mL,and its purity was>90%.The monoclonal antibodies prepared in this study have good biological characteristics,which provides a new method for the research and rapid diagnosis of PCV2.

关 键 词：猪圆环病毒2型 REP蛋白 单克隆抗体 制备 

分 类 号：S811.5[农业科学—畜牧学]