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利用重组质粒pCZS转化大肠杆菌,制备出人重组铜-锌超氧化物歧化酶(rhCu-Zn SOD),进一步建立了3株鼠抗rhCu-Zn SOD的单克隆抗体(McAb)杂交瘤细胞株.经鉴定,它们分泌的McAb分别属于IgG_1和IgG_2a亚类,能特异性地识别人Cu-Zn SOD,与pCZS空载菌蛋白无交叉反应.将此McAb制备了免疫亲和层析柱,从大肠杆菌的粗提液中纯化出rhCu-Zn SOD,十二烷基磺酸钠—聚丙烯酰胶凝胺电泳(SDS-PAGE)显示单一条带,纯度>98%,回收率50%,酶比活达10 971.3 U/mg蛋白,rhCu-Zn SOD McAb的制备为Cu-Zn SOD产品的纯化、分析及作用机理研究等提供了有力的工具.
Recombinant plasmid pCZS was transformed into Escherichia coli to prepare human recombinant copper-zinc superoxide dismutase (rhCu-Zn SOD), and further established three monoclonal antibody (McAb) hybridoma cell strains which are anti-rhCu-Zn SOD. The McAbs secreted by the McAbs were identified as IgG_1 and IgG_2a subclasses, respectively, which could specifically recognize human Cu-Zn SOD and had no cross-reaction with pCZS no-load protein.The McAbs were prepared by immunoaffinity chromatography RhCu-Zn SOD was purified from the crude extract of Bacillus, SDS-PAGE showed a single band with purity> 98% and recovery of 50% Activity of 10 971.3 U / mg protein, the preparation of rhCu-Zn SOD McAb provides a powerful tool for the purification, analysis and mechanism study of Cu-Zn SOD products.