抗MS_2多聚酶单克隆抗体的制备及鉴定

应用MS_2-X蛋白(含MS_2多聚酶的98个氨基酸,乙型肝炎X抗原的145个氨基酸)免疫小鼠,取其脾细胞与Sp2/0细胞融合后,经阳性克隆检测,克隆化及体外传代培养,共筛选出了2株抗MS_2多聚酶的单克隆抗体,这在国内尚未见报道。经ELISA及Western Blot实验证实,此2株单克隆抗体只能与MS_2多聚酶发生反应,而不能与乙型肝炎X抗原和正常细菌蛋白发生反应,其抗体亚类为IgG_1,腹水效价为1∶32000～1∶64000。应用抗MS_2单克隆抗体,可以建立亲和层析技术,高效快速地将基因工程表达的MS_2多聚酶—目的蛋白融合蛋白,从大量的细菌蛋白中纯化出来,这对深入研究目的蛋白的结构与功能具有十分重要的作用。

Preparation and Characterization of Monoclonal Anti-MS2 Polymerase Antibodies

By fusing Sp2/0 cells with spleen cells from BALB/c mice immunized with MS2-X fusion protein ( MS2 polymerase, 98 amino acids, hepatitis B virus X antigen, 145 amino acids ) , two clones of monoclonal anti-MS2 polymerase antibodies were generated after screening, cloning, subcloning, and passaging. It was confirmed by ELISA and Western blot that both the two clones might react with MS2-X protein and sonicated E. coli C600 proteins containing MS2 polymerase, but they might not react with normal E. coli C600 proteins and MY-X protein (hepatitis B virus X antigen non-fusion protein). Both the two clones were of IgG isotype with ascitic titres ranging from 1:32000 to 1:64000. Monoclonal anti-MS2 polymerase antibodies might be used to purify recombinant MS2 fusion proteins from E. coli proteins by affinity chromatogra-phy.