鼠源抗人乳腺癌MRP1/ABCC1噬菌体Fab抗体库的构建  

Construction of a Murine Phage Fab Library Against MRP1/ABCC1 of Human Breast Carcinoma

作　　者：张学梅[1] 赵彩红[1] 李俊[1] 罗晓[2] 蒋葵[2] 

机构地区：[1]大连大学医学院,中国辽宁大连116622 [2]大连医科大学附属第二医院肿瘤科,中国辽宁大连116027

出　　处：《生命科学研究》2014年第4期338-343,共6页Life Science Research

基　　金："十一五"国家科技支撑计划项目(2009BAK61B04)

摘　　要：鼠源抗人乳腺癌多药耐药相关蛋白1(multidrug resistance-associated protein 1,MRP1/ABCC1)噬菌体Fab抗体库的构建首先是利用乳腺癌组织匀浆液作为免疫原免疫BALB/c小鼠,成功免疫后提取小鼠脾组织的总RNA逆转录为cDNA。然后设计小鼠IgG各亚型的特异性Fab引物,PCR扩增Fd和L基因并依次连接到噬菌体载体pComb3中。重组体pComb3-Fab转化至E.coli XL1-Blue菌中,最后经辅助噬菌体VCSM13超感染,成功构建噬菌体Fab抗体库。以化学合成的MRP1/ABCC1膜表位10肽为抗原进行3轮淘选、富集,对淘选后获得的各级噬菌体抗体库进行ELISA检测和鉴定。成功获得的抗MRP1/ABCC1-10肽三级噬菌体Fab抗体库将为抗人乳腺癌MRP1/ABCC1 Fab抗体的制备提供保障。The murine phage Fab antibody library against multidrug resistance-associated protein 1（MRP1/ABCC1） of human breast carcinoma was constructed successfully. The BALB/c mice were immunized with tissue homogenate of human breast carcinoma. Total RNA was extracted from immune mice splenocytes and reversely transcribed into cDNA. The Fab genes were amplified by PCR with specific primers, and the Fd and light chain genes were inserted into the phagemid vector pComb3 separately. The pComb3-Fab was transformed into E.coli XL1-Blue, then the clones were infected by the helper phage VCSM13. Three rounds of enrichment and panning were performed to obtain Fab antibody library against synthetic MRP1/ABCC1-10 peptide, then the three Fab antibody libraries were identified by ELISA. The third murine phage Fab library against MRP1/ABCC1-10 peptide of human breast carcinoma was obtained successfully, which might provide the chance for obtaining the Fab antibody against MRP1/ABCC1 of human breast carcinoma.

关 键 词：FAB抗体 多药耐药相关蛋白1(MRP1 ABCC1) 乳腺癌 

分 类 号：R392.12[医药卫生—免疫学]