用体外致敏联合EB病毒转化技术构建抗肝癌的人源Fab噬菌体抗体库  

Construction of phage antibody library using sensitized in vitro B-lymphocytes of liver cancer patients

作　　者：水漩[1] 李官成[1] 李跃辉[1] 黄建[1] 赵艳[1] 

机构地区：[1]中南大学肿瘤研究所免疫室,湖南长沙410078

出　　处：《西安交通大学学报（医学版）》2007年第4期364-367,共4页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：湖南省卫生厅科研基金重点资助项目(No.A2003001)

摘　　要：目的构建抗肝癌的人源Fab噬菌体抗体库。方法体外致敏并用EB病毒(EBV)转化肝癌患者的外周血单个核细胞(PBMC)。RT-PCR扩增人抗体轻链和重链Fd段基因,将抗体基因分别与载体pComb3连接后,电转化大肠杆菌XLI-Blue,构建噬菌体呈现型Fab抗体库。结果经EBV转化的4例肝癌患者PBMC,ELISA检测均有抗肝癌抗体产生;经PCR扩增出13条轻链基因片段(κ、λ)和28条重链Fd基因片段(γ);电转化构建的噬菌体抗体库库容为1.7×107puf/mL;Fab基因的重组率约为100%。结论用体外致敏法结合噬菌体抗体库技术,成功构建了全人源抗肝癌Fab组合抗体文库。To construct human phage antibody library against hepatoma carcinoma. Methods Peripheral blood mononuclear cells （PBMCs） of patients with liver cancer were sensitized in vitro and transformed by Epstein-Barr virus （EBV）. The genes of light chain and Fd of antibodies were amplified by RT-PCR. Fab genes were cloned into vector pComb3 and transformed into E. coli XLI-Blue by electroporation to construct the Fabdisplaying phage antibody library. Results ELISA detection showed that 4 liver cancer patients＇ B cells transformed by EBV could produce specific antibodies to hepatoma carcinoma cell. Totally 13 types of light chain genes and 28 types of Fd genes were obtained by RT-PCR. The capacity of the primary phage library was 1.7 × 107pfu/mL. The percentage of recombinant clones was about 100%. Conclusion A human phage antibody library has been constructed successfully by means of EBV transformation technique.

关 键 词：EB病毒转化 噬菌体抗体库 肝癌 FAB抗体 

分 类 号：R739.63[医药卫生—肿瘤]