摘要
目的 对抗胃癌单链抗体免疫毒素基因重组质粒进行表达 ,进而获得有活性的单链抗体免疫毒素。方法 应用IPTG诱导大肠杆菌表达系统进行单链抗体免疫毒素基因重组质粒的表达 ,并对产生的不溶性包涵体进行纯化 ,经变性、复性后 ,观察其生物学活性。结果 诱导表达了约 6 6KD的蛋白 ,分子量符合预计大小 ,以包涵体的形式存在于菌体中。经包涵体纯化、变性、复性 ,使其折叠为正确的空间结构 ,经检测可与胃癌细胞MGC80 3特异结合 ,并有一定的细胞毒性。结论 获得了可与胃癌细胞系MGC80 3特异结合 。
Objective To obtain the active anti-carcinoma ScFv immunotoxin.Methods In E.coli the anti-carcinoma ScFv immunotoxin recombinant plasmid was induced by IPTG to express the ScFv immunotoxin in inclusion bodies,and the latter were succesively purified、denatured and renatured.By competitive inhibition ELISA and colorimetric MTT assay,the activity of anti-carcinoma ScFv immunotoxin was determined.Results The anti-carcinoma ScFv immunotox recombinant plasmid expressed the ScFv immunotoxin in inclusion.Its molecular weight was about 66KD.The ScFv immunotoxin in inclusion was refolded to form correct structure through denaturation and renaturation showed specific and cytotoxic to gastric cancer cell line MGC803.Conclusions The resultant anti-carcinoma ScFv immunotoxin is of strict specificity and cytotoxicity to gastric cancer cell line MGC803.
出处
《宁夏医学杂志》
CAS
2001年第5期259-261,共3页
Ningxia Medical Journal
