抗癌胚抗原微型抗体基因在昆虫细胞中的表达  

Expression Gene of Miniantibody to Carcionembryonic Antigen in Insect Cells

作　　者：杨卫东[1] 李彪[1] 朱承谟[1] 贺华君[2] 杨冠珍[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》2001年第1期4-7,共4页Acta Universitatis Medicinalis Secondae Shanghai

基　　金：国家自然科学基金!( 3 9770 2 3 1);上海市启明星计划!( 98Q14 0 11)

摘　　要：目的获得生物活性更高的癌胚抗原 (CEA)微型抗体用于放免显像。　方法将抗癌胚抗原微型抗体基因插入昆虫杆状病毒供体质粒pFastBacHTb中 ,经大肠杆菌DH10Bac体内转座 ,产生重组杆状病毒BacHT -VH-L ,将其转染粉纹夜蛾 (Tn - 5B1- 4)细胞 ,经扩增后在细胞内进行表达。　结果SDS -PAGE分析结果表明 ,在Tn- 5B1- 4细胞中表达产生一条特异性蛋白质 ,其分子量为 2 6kd左右 ;以Westernblot分析表明 ,该特异条带即为VH-L蛋白。RIA表明重组杆状病毒表达产生的VH -L蛋白能特异性的结合CEA ,较大肠杆菌表达物活性更高。结论昆虫细胞表达可制备生物活性更高的癌胚抗原微型抗体。Objective To Produce miniantibody which can bind carcionembryonic antigen (CEA) better, being suitable for radioimmunoimaging. Methods The gene of miniantibody to CEA was inserted into the donor plasmid pFastBacHTb. Then the donor plasmid was transformed into the E.coli DH10Bac, in which transposition took place. The recombinant bacmid DNA, which contained the miniantibody gene, was extracted from the E.coli DH10Bac, and transfected the Tn-5B1-4 cells. The recombinant baculovirus were obtained. After amplification, the recombinant baculovirus infected the Tn cells and the gene of miniantibody was highly expressed in Tn cells. Results SDS-PAGE analysis revealed that the molecular weight of miniantibody was 26 kd. It was shown it possessed ability to bind to its specific antigen of CEA by RIA and was better than the protein expressed in E.coli. Conclusion It is a valid method to produce miniantiboy which has high ability in binding CEA by expression in Tn cells.

关 键 词：癌胚抗原 微型抗体 昆虫杆状病毒 基因表达 肿瘤 

分 类 号：R73-3[医药卫生—肿瘤]