抗猪圆环病毒2型病毒样颗粒单克隆抗体的制备及鉴定  

Preparation and identification of monoclonal antibodies against porcine circovirus type 2 virus-like particle

作　　者：侯强[1] 侯绍华[1] 贾红[1] 姜一曈[1] 鑫婷[1] 李明[1] 陈青[2] 朱鸿飞[1] HOU Qiang;HOU Shao-hua;JIA Hong;JIANG Yi-tong;XIN Ting;LI Ming;CHEN Qing;ZHU Hong-fei(Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China;Key Laboratory of Urban Agriculture （North） of Ministry of Agriculture China, Beijing University of Agriculture, Beijing 102206, China)

机构地区：[1]中国农业科学院北京畜牧兽医研究所,北京100193 [2]北京农学院农业部都市农业(北方)重点实验室,北京102206

出　　处：《中国预防兽医学报》2018年第4期325-329,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家重点发展计划项目(2017YFD0502302);北京市教委科技计划面上项目(KM201410020002);中国农业科学院科技创新工程(ASTIP-IAS-11)

摘　　要：为制备能够特异性识别猪圆环病毒2型(PCV2)病毒样颗粒(VLP)的单克隆抗体(MAb),本研究利用杆状病毒表达系统分别制备了具有VLP结构的核衣壳蛋白(Cap)和去掉核定位信号肽的tCap蛋白,以前者为免疫原,后者为筛选抗原制备了一株杂交瘤细胞(1F6),通过western blot、间接免疫荧光和透射电镜对其生物学特性进行鉴定。结果显示,1F6 MAb与PCV2 VLP具有良好的特异性反应,同时能够特异性识别天然PCV2和线性结构的Cap蛋白。本研究制备的MAb为PCV2抗原表位的鉴定和PCV2 VLP定量方法的建立提供了工具。In order to prepare monoclonal antibodies against porcine circovirus type （PCV2） virus-like particle （VLP）, Cap protein with the PCV2 VLP structure （cap） and a truncated Cap protein （tCap） without the N-terminal nuclear localization signal were expressed with recombinant baculovirus system and purified from insect ceils respectively. A hybridoma cell （1F6） was prepared by using Cap protein with VLP structure as immunogen and tCap as screening antigen. The biological characteristics of 1F6 MAb were identified by western blot, IFA indirect immunofluorescence and transmission electron microscopy. The results showed that 1F6 MAb could specifically recognize PCV2 VLP, PCV2 virion and linear Cap protein. 1F6 MAb provides a tool for identification of PCV2 epitopes and establishment of a PCV2 VLP quantitative method.

关 键 词：猪圆环病毒2型 单克隆抗体 核衣壳蛋白 病毒样颗粒 

分 类 号：S852.65[农业科学—基础兽医学]