摘要
用纯化的猪繁殖与呼吸综合征病毒(PRRSV)A06株病毒抗原免疫Balb/c小鼠,取免疫小鼠脾细胞与SP2/0骨髓瘤细胞进行细胞融合。经间接ELISA方法筛选,有限稀释法克隆,获得4株稳定分泌特异性单克隆抗体的杂交瘤细胞株,分别命名为3D12、4E10、6E2和7G6。其细胞培养上清效价分别为1∶256、1∶512、1∶512和1∶256,小鼠腹水效价分别为1∶1.024×106、1∶2.048×106、1∶1.024×106和1∶2.048×106。亚型鉴定表明,3D12、4E10、6E2和7G6分别为IgG2b、IgG2b、IgG1和IgG2a。ELISA检测结果显示,4种PRRSV单克隆抗体仅与PRRSV反应,不与其他病毒反应,表明均为抗PRRSV的型特异性单克隆抗体。对单克隆抗体腹水液进行IgG提取纯化,SDS-PAGE电泳验证,仅出现两条清晰条带,无杂带出现,证明纯化效果较好。这4种单克隆抗体的获得,为建立PRRSV检测方法提供了强有力的工具。
The paper was to obtain monoclonal antibodies （McAbs） against porcine reproductive and respiratory syndrome virus（PRRSV） . McAbs 3D12,4E10,6E2 and 7G6 against PRRSV were produced by fusing SP2/0 myeloma cells with spleen cells of Balb/e mice immunized with purified PRRSV. The indirect ELISA results showed that the antibody titers of cell supernatants were 1 ： 256, 1 ： 512, 1 ： 512 and 1 ： 256,and the titers of ascites were 1 ： 1.02×10^6,1 ： 2. 048×10^6,1 ：1. 024×10^6 and 1 ： 2. 048×10^6,respectively. The antibody subclass analysis showed that 3D12 and 4E10 were IgG2b, 6E2 was IgGl,and 7G6 was IgG2a. The result of ELISA showed that all the McAbs reacted with PRRSV but not with other viruses. SDS PAGE analysis showed that the purified results of all the McAbs in ascites were good. The MeAbs are useful tools for developing diagnostic techniques of PRRSV.
出处
《动物医学进展》
CSCD
北大核心
2009年第12期1-4,共4页
Progress In Veterinary Medicine
基金
国家质检总局科技项目(2008IK262-2)
深圳出入境检验检疫局项目(SZ2007038)
关键词
猪繁殖与呼吸综合征病毒
单克隆抗体
生物学特性
Porcine reproductive and respiratory syndrome virus
monoclonal antibody
biological characteristics
