CEA微型抗体载体的构建及在大肠杆菌中的高效表达  

Construction of Miniantibody of Carcinoembryonic Antigen and High Expression in E. coli

作　　者：杨卫东[1] 李彪[1] 朱承谟[1] 廖文韬[2] 吴祥甫[2] 

机构地区：[1]上海第二医科大学瑞金医院核医学科,上海200025 [2]中国科学院上海生物化学研究所

出　　处：《上海第二医科大学学报》2000年第3期199-202,共4页Acta Universitatis Medicinalis Secondae Shanghai

基　　金：国家自然科学基金 !(39770 2 31 );上海市启明星计划! (98Q1 4 0 1 1 )

摘　　要：目的制备适合放免显像的抗癌胚抗原微型抗体。　方法在已获得的抗癌胚抗原重链可变区 (VH)及轻链可变区 (VL)基因的基础上将重链可变区 (VH)与轻链可变区 (VL)基因直接连接制成微型抗体基因 ,并在大肠杆菌进行表达。　结果大肠杆菌高效表达了微型抗体C端 6×His的融合蛋白 ,表达量达菌体总蛋白的 15 % ,SDS -PAGE和Western印迹显示表达物分子量为 2 6kd与基因编码蛋白的理论推算值相符 ,RIA表明表达物与CEA特异性结合。Objective Construction miniantibody of carcinoembryonic antigen for radioimmunoimaging. Methods The heavy chain variable and light chain variable genes encoding the murine monoclonal antibody E 7B 10 with specificity for carcinoembryonic antigen which have been constructed were linked directly and expressed in E.coli. Results The construction which conclude 6×His at C-terminal was expressed in E.coli with high level reaching 15% of the total bacteria protein, and chara cterized by SDS-PAGE, Western blot, the expression product had the CEA-binding ability in RIA.Conclusion It is a valid method to produce miniantibody of carcinoembryonic antigen by expression in E.coli.

关 键 词：放射损度显像 抗癌胚抗原微型抗体 制备 

分 类 号：R730.3[医药卫生—肿瘤]