猪肺炎支原体168株特异性单抗的制备及鉴定

Preparation and Identification of Monoclonal Antibodies against Mycoplasma hyopneumoniae Strain 168

将猪肺炎支原体168株全菌蛋白免疫小鼠,取脾细胞与SP2/0骨髓瘤细胞进行融合,3次亚克隆后,获得了4株针对168株全菌蛋白的单抗,分别将其命名为2F5、2G7、4F5、5E9。Western-blot检测结果表明,2G7、4F5与猪肺炎支原体具有特异性反应条带,不与猪鼻支原体、大肠杆菌反应。亚型鉴定结果表明,两株特异性单抗(2G7、4F5)的亚型属IgG1,轻链为κ型,2F5、5E9亚型属IgG2a,轻链为κ型。间接免疫荧光结果表明,4株单抗均与猪肺炎支原体168株有特异性荧光反应,特异性单抗的获得为猪肺炎支原体的检测及机理研究奠定基础。

Balb/c mice were immunized with Mycoplasma hyopneumoniae （Mhp） strain 168, and the splenocytes of the immu- nized mice were fused with SP2/O cells. After subcloning for 3 times, four hybridoma clones which produced Monoclonal Antibodies （McAbs） steadily were screened by ELISA, named 2F5, 2G7, 4F5, and 5E9. The result of Western -blot indicated that 2G7 and 4F5 both strongly specifically reacted with the Mhp strain 168. Both of the McAbs （2G7 and 4F5） belonged to IgG1 isotype, and their light chains were K type. However, 2F5 and 5E9 belonged to IgG2a isotype, and their light chains were also K type. They all reacted strongly with Mhp strain 168 in immunofluorescence assay （IFA）. The McAbs could be used for the detection and pathogenic mecha- nism research of Mhp.