关键词: 胃肿瘤;人抗体，肿瘤;噬菌体

    摘 要:目的 利用噬菌体表面展示技术，构建人源性胃癌抗体IgG1Fab噬菌体表面展示文库. 方法 从胃癌手术患者胃一级站淋巴结中提取总RNA，反转录成cDNA后，用PCR扩增人IgG1Fd段及κ轻链，并克隆至噬菌粒载体pCOMB3中，转化宿主菌株XL1-Blue，以辅助噬菌体M13K07超感染噬菌粒文库，构建成人源性胃癌抗体IgG1Fab噬菌体表面展示文库. 结果 得到一个含克隆数为1.2×106 ，实际滴度约为2.56×1015 pfu·L-1 的IgG1Fab片段的噬菌体表面展示文库. 结论 IgG1Fab片段的噬菌体表面展示文库的成功构建，为下一步利用亲和富集筛选技术，获得具有胃癌表面抗原结合活性的融合抗体及可溶性抗体奠定了基础.

    Construction of phage display library of IgG1Fab antibodies from lymph nodes of gastric cancer pa-tients

    HAN Zhe-Yi，HE Feng-Tian，CHEN Bao-Jun，QIAO Tai-Dong，WANG Hai-Yan，HAN Ying，ZHENG Jian-Yong，WU Kai-Chun，FAN Dai-Ming

    1 Institute of Gastroenterology of Chinese PLA，2 Department of Gastrointestinal Surgery，Xijing Hospital，Fourth Military Medical University，Xi'an710033，China

    Keywords:stomach neoplasms;antibodies，neoplasms;bac-teriophages

    Abstract:AIM To construct a phage display library of hu-man IgG1Fab antibodies from lymph nodes of gastric cancer patients using phage display technique.METHODS Total RNA was isolated from lymph nodes of gastric cancer pa-tients.Fd fragment and kappa light-chain were amplified by RT-PCR and cloned into the phagemid pCOMB3and ex-pressed as a fusion protein with phage M13PⅢin E.coli XL1-Blue.In the presence of help phage M13K07 ......