抗猪圆环病毒2型Cap蛋白中和性单克隆抗体的制备及鉴定  

Preparation and identification of neutralizating monoclonal antibody to the recombinant capsid protein of porcine circovirus type 2

作　　者：黄立平[1] 刘长明[1] 危艳武[1] 张朝霞[1] 陆月华[1] 郭龙军[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/猪传染病研究室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2009年第2期132-136,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：引进国际先进农业科学技术(948)项目(2006-Z6);国家科技支撑计划项目(2006BAD06A07)

摘　　要：以纯化的杆状病毒表达的重组猪圆环病毒2型(PCV2)Cap蛋白(PCV2-rCap)免疫BALB/c小鼠,利用淋巴细胞瘤杂交技术获得了5株稳定分泌抗PCV2-rCap蛋白的杂交瘤细胞株,分别命名为1D2、2E8、3A10、5F2和6F10。这5株杂交瘤细胞诱导同品系小鼠产生的腹水抗体效价分别为1∶2048000、1∶512000、1∶1024000、1∶512000、1∶1024000;其中1D2、2E8、5F2和6F10单抗亚类为IgG2a,仅3A10为IgG1;2E8、3A10、5F2和6F10单抗轻链为λ型,仅1D2轻链为κ型。Western blot结果显示,这5株单抗中2E8、3A10、5F2和6F10可与自然PCV2-Cap发生反应,而1D2无反应。IPMA和抗原捕获ELISA结果表明,1D2单抗可与PCV2病毒蛋白发生反应,而与PCV1无交叉反应;病毒中和试验证实,1D2单抗具有中和活性,是一株针对PCV2-Cap蛋白中和表位的单抗。这些单抗的制备为PCV2抗原表位分析及分子诊断提供了技术手段。BALB/c mice were immunized with the purified recombinant capsid （rCap） protein of porcine circovirus type 2 （PCV2） prepared from baculovirus expression system. Five monoclonal antibodies （McAb） designated 1D2, 2E8, 3A10, 5F2, and 6F10, were obtained. These ascites titers of the McAbs were 1：2 048 000, 1：512 000, 1：1 024 000, 1：512 000 and 1：1 024 000 for 1D2, 2E8, 3A10, 5F2 and 6F10, respectively. All McAbs belonged to IgG2a subclass, except for 3A10 which belonged to IgG1. The McAbs had λ chain for 2E8, 3A10, 5F2 and 6F10, and κ chain for 1D2. Western blot analysis showed that all McAbs but 1D2, were able to react with native PCV2-Cap; However, 1D2 was able react with PCV2 in immunoperoxidase monolayer assay （1PMA） and antigen capture ELISA. The McAb Of 1D2 was confirmed to have the neutralizing ability to PCV2 in the neutralization test. These McAbs would be useful tools for molecular diagnostic and antigen epitope analysis of the virus.

关 键 词：猪圆环病毒2型 重组CAP蛋白 中和活性 单克隆抗体 

分 类 号：S852.659[农业科学—基础兽医学]