胰腺癌人源噬菌体单链抗体库的构建及初步筛选  

Construction and screening of single-chain antibody library of pancreatic cancer

作　　者：何光志[1] 田维毅[1] 高英[2] 王平[1] 王文佳[1] 黄高[1] 蔡琨[1] 安传伟[1] 

机构地区：[1]贵阳中医学院,贵州贵阳550002 [2]遵义医学院附院,贵州遵义563003

出　　处：《广东农业科学》2011年第7期158-160,共3页Guangdong Agricultural Sciences

基　　金：贵州省科学技术基金(黔科合J字[2010]2262号)

摘　　要：提取胰腺癌患者的外周淋巴细胞总RNA,以RT-PCR技术分别扩增出人源抗体H链和L链的可变区基因,采用SOE-PCR法将VH和VL片段随机拼接成scFv片段,然后将scFv片段克隆至pCANTAB5E载体,并电转化至感受态TG1菌,经辅助噬菌体M13K07拯救,获得胰腺癌人源噬菌体单链抗体库。结果表明,从外周血淋巴细胞中获取可变区基因,利用噬菌体抗体库技术制备人源抗胰腺癌单链抗体的策略是可行的。Total RNA was abstracted from peripheral blood lymphocytes of patients suffering from pancreatic cancer , and was used to amplify VH and VL gene by RT-PCR. VH and VL were joined by SOE-PCR to form the single chain variable fragment （ScFv） gene. These SeFv fragments were cloned into the phagemid pCANTABSE and the recombinant phagemids were ransformed to susceptible E.coli TGI by roporation. After rescuing by helper phage M13K07, a natural single-chain antibody library for pancreatic cancer was successfully constructed. This study showed that in the construction of engineering antibodies against human pancreatic cancer, the strategy of phage display technique and of RT-PCR from peripheral blood lymphocytes mRNA may provide an alternative approach.

关 键 词：胰腺癌 人源噬菌体单链抗体 构建与筛选 

分 类 号：Q785[生物学—分子生物学]