肺癌患者噬菌体抗体库的构建及抗肺癌单抗的筛选  

Construction of phage antibody libraries of patients with lung cancer and selection of human monoclonal antibodies against lung cancer

作　　者：胡建林[1] 郭先健[2] 胡义德[2] 黄桂君[2] 李淑平[2] 陈维中[2] 

机构地区：[1]第三军医大学西南医院呼吸科,重庆400038 [2]第三军医大学新桥医院呼吸科,重庆400037

出　　处：《免疫学杂志》2002年第4期299-302,共4页Immunological Journal

摘　　要：目的　探索从人体肿瘤局部引流淋巴结扩增免疫球蛋白基因构建噬菌体抗体库及从中筛选抗肿瘤单抗的可行性。方法　从肺癌患者肺门淋巴结提取总RNA ,经逆转录 聚合酶链反应扩增免疫球蛋白重链Fd段和κ链基因 ,以pcomb3H为载体 ,构建噬菌体抗体库 ,以 2株肺癌细胞对噬菌体抗体库进行 6轮亲和筛选 ,再以人成纤维细胞进行 2轮吸附 ,以筛选出抗肺癌细胞抗体。以细胞ELISA法初步鉴定抗体活性。结果　构建成噬菌体抗体库 ,库容为 7.5× 10 7个克隆 ,κ链和Fd段基因与载体DNA的重组率分别为 10 0 % (10 / 10 )和 90 % (9/ 10 )。通过亲和筛选和吸附从中获得了能与肺癌细胞结合的人单抗。结论　从人体肿瘤局部引流淋巴结扩增免疫球蛋白基因构建噬菌体抗体库并从中筛选抗肿瘤单抗是可行的 ,以肿瘤细胞作抗原从抗体库中筛选抗肿瘤抗体是可取的。Objective To investigate the feasibility of construction of phage antibody libraries by amplifying immunoglobulin genes from lymph nodes of patients with cancer and selection of monoclonal antibodies against cancer from the libraries. Methods Immunoglobulin heavy chain Fd genes and κ chain genes were amplified by RT PCR from regional lymph nodes of patients with lung cancer. Phage antibody library was constructed by cloning Fd genes and κ chain genes into phagemid pComb3H. Six rounds of affinitive selection with lung cancer cells as antigens and 2 rounds of absorption with human fibroblasts were carried out. Specificities and sensitivities of selected antibodies were determined by cell ELISA. Results A phage antibody library of 7.5×10 7 clones was constructed. In the library, 100%(10/10) of recombined DNAs contain κ chain gene and 90%(9/10) contain Fd gene. After 6 rounds of selection against lung cancer cells and 2 rounds of absorption with human fibroblasts, antibodies against lung cancer cells were acquired. Conclusions It is feasible to construct phage antibody libraries by amplifying immunoglobulin genes from lymph nodes of patients with cancer. It is desirable to select monoclonal antibodies against cancer from the libraries with cancer cells as antigens.

关 键 词：噬菌体抗体 人源性抗体 肺肿瘤 抗肿瘤单克隆抗体 肺癌 

分 类 号：R734.2[医药卫生—肿瘤]