全人源肝癌噬菌体单链抗体基因的克隆及活性分析  

Cloning and analyzing the function of the gene of human phage antibodies with single - chain variable fragment specific to hepatocellular cancer

作　　者：薛国柱[1] 窦科峰[1] 赵爱志[1] 药立波[2] 

机构地区：[1]第四军医大学西京医院肝胆外科,陕西西安710032 [2]第四军医大学基础部生物化学与分子生物学教研室,陕西西安710032

出　　处：《临床肝胆病杂志》2005年第4期218-219,共2页Journal of Clinical Hepatology

基　　金：国家自然科学基金资助项目(No.30371399)

摘　　要：从噬菌体单链抗体库中筛选克隆全人源肝癌抗体基因并进行活性鉴定。PCR鉴定阳性重组菌中人肝癌ScFv的插入率,以肝癌细胞SMMC-7721为抗原对所建抗体库进行4轮“吸附-洗脱-扩增”的亲和筛选。将筛选后的ScFv采用ELISA法鉴定其与人肝癌细胞的结合活性。ScFv基因插入率为70%。在亲和筛选过程中,肝癌噬菌体单链抗体得到富集,收获率逐轮提高,第4轮为第一轮的381倍。利用噬菌体抗体库技术筛选出了肝癌噬菌体单链抗体,且筛选后的抗体片段与人肝癌细胞有特异性的结合活性。Objective Cloning and characterizing the gene of human phage antibodies with single - chain variable fragment （ScFv） specific to hepatoeellular cancer. Methods The recombinant phages inserted with ScFv were determined by PCR. The recombinant phages were panned by human hepatocellular cancer cell line of SMMC - 7721. After 4 rounds of biopanning, the specificity of ScFv from the monoclonal bacteria was determined by ELISA. Results The rate of recombinant bacteria inserted with ScFv was 70%. From the 1st round to the 4th round of the panning, the number of the eluted phages had increased 381 times. Conclusion： Acquired phage antibodies with ScFv specifically eombined with hepatocellular cancer by the phage antibody panning.

关 键 词：噬菌体抗体库 抗体 单克隆 肝肿瘤 

分 类 号：R735.7[医药卫生—肿瘤]