抗人大肠癌P-gp_(185)Fab抗体的制备  

Obtainment of the Fab Antibody Against P-gp_(185) in Human Colorectal Cancer

作　　者：李俊[1] 蔺娜[2] 卞威[3] 张学梅[1] 

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一医院药剂科,中国辽宁大连116021 [3]沈阳市第一人民医院神经外科,中国辽宁沈阳110041

出　　处：《生命科学研究》2013年第6期512-516,共5页Life Science Research

基　　金："十一五"国家科技支撑计划项目(2009BAK61B04)

摘　　要：针对人大肠癌P-gp(P-glycoprotein),获得用于研究其功能的抗某一线性表位的单克隆Fab抗体.以原核表达的P-gp185为抗原,对前期所构建的库容量为2.47×106cfu的抗人大肠癌Fab抗体库进行5轮的淘选和富集.ELISA法鉴定获得的各级抗P-gp185Fab抗体库.利用化学合成的膜表位P-gp10肽(ANDAAQVKGA)从五级库中获得表达阳性Fab抗体的克隆及该Fab抗体的基因序列.结果显示,成功富集、淘选获得各级抗人大肠癌P-gp185Fab抗体库;成功获得抗线性表位P-gp10肽的阳性克隆——24号菌株及其表达的Fab抗体基因序列.该Fab抗体的后续制备有望用于大肠癌P-gp功能的研究,该方法也为研究其它目标蛋白的功能提供了有价值的线索.To obtain the Fab antibody against a linear epitope of P-gp （P-glycoprotein） in human colorectal cancer. Five rounds of panning and enrichment were performed to pick the specific Fab antibodies against P-gp185 from the Fab antibody library with 2.47×10^6 efu constructed before. Each round of Fab antibody library against P-gp185 was identified and the optimal clone against P-gp10-peptide from the fifth round library was selected by ELISA as well. After panning, both the Fab antibody library against P-gp185 and the optimal clone （No.24） against P-gplO-peptide were obtained successfully. Under the obtainment the DNA sequence of the No.24, preparation of Fab antibody against P-gp10-peptide might be used to study the function of P-gp in human colorectal cancer, and the procedure might provide a valuable clue to explore the function of any other objective proteins.

关 键 词：P-GP FAB抗体 大肠癌 

分 类 号：R392.12[医药卫生—免疫学]