抗猪(嗜血)支原体MSG1蛋白单克隆抗体的制备及鉴定

Preparation and identification of monoclonal antibodies against MSG1 protein of Mycoplasma suis

以原核表达纯化的猪(嗜血)支原体MSG1蛋白免疫BALB/c小鼠,运用细胞融合技术筛选分泌针对MSG1蛋白抗体的融合细胞。通过间接ELISA方法筛选获得2株能稳定分泌抗体的融合细胞株,分别命名为1A7和3G6,Western blot结果证明这2株细胞分泌的抗体能够与重组MSG1蛋白发生特异性反应。细胞上清和腹水中的ELISA抗体效价分别为1∶4 096、1∶1 024和1∶1 638 400、1∶51 200,其单抗亚类鉴定均属于IgG1,轻链为κ型。抗原识别位点分析结果表明,2株单抗所识别的抗原位点相同。猪(嗜血)支原体MSG1蛋白特异性单克隆抗体的制备成功,为制备免疫诊断试剂盒和致病机制的研究奠定了基础。

The monoclonal antibodies （MAbs） were prepared by fusing mouse myloma ceils （SP2/0） with spleen cells from BALB/c mice immunized with purified recombinant MSG1 protein of Mycoplasma suis. Two hybridoma cell lines secreting MAbs against MSG1 protein were screened by indirect enzyme-linked immunosorbent assay （ELISA） and named as 1 A7 and 3 G6, respectively. Western-blot results showed that the two MAbs reacted specifically with recombinant MSG1 protein. Their ELISA titers in supernatant and asicite were 1 ： 4096, 1 ： 1024 and 1 ： 1638400, 1 ： 51200, respectively. The isotypes of 1A7 and 3G6 both belong to IgG1/κ and the additive ELISA showed that the two MAbs reacted with the same antigen determinant. So, we concluded that the specific anti-MSG1 protein MAbs were developed and these MAbs may be useful in the development of detection methods and provide a basis for investigating the pathogenicity of M. suis.