丙型肝炎病毒核心蛋白人源单链可变区抗体的筛选与鉴定  

Screening and characterization of human phage antibody to hepatitis C core antigen

作　　者：钟彦伟[1] 成军[1] 施双双[1] 夏小兵[1] 王刚[1] 杨继珍[1] 陈菊梅[1] 

机构地区：[1]北京解放军第302医院传染病研究所基因治疗研究中心,100039

出　　处：《中华肝脏病杂志》2001年第4期217-219,共3页Chinese Journal of Hepatology

基　　金：国家自然科学基金资助项目(39900130)

摘　　要：目的筛选、鉴定抗丙型肝炎病毒（ＨＣＶ）核心蛋白的人源单链可变区抗体（ＳｃＦｖ）。方法采用噬菌体表面展示技术，以重组的ＨＣＶ核心蛋白为包被抗原，从噬菌体单链可变区抗体库中经过３轮“吸附－洗脱－扩增”筛选过程，获得抗原结合活性较强的ＨＣＶ核心蛋白特异性人源单链可变区抗体片段阳性克隆，并对其进行免疫学及核苷酸序列测定。结果筛选得到的ＳｃＦｖ片段具有抗ＨＣＶ核心蛋白的特异性，基因序列分析结果表明符合人源单链可变区抗体基因序列的结构特征。结论利用噬菌体抗体库技术，成功获得ＨＣＶ核心蛋白的特异性人源单链可变区抗体的编码基因。Objective To screen and characterize human phage antibody (ScFv) against hepatitis C core antigen. Methods The recombinant phages were panned by core antigen that was coated in a microtiter plate. After three rounds of biopanning, 48 clones were determined specific to core antigen. The specificity of each ScFv colone was determined by ELISA. The coding gene for HCV protein ScFv has been sequenced. Results Phage antibody for HCV core protein had a specific combination character with core antigen of hepatitis C virus. The DNA sequence data showed that the ScFv gene included 774 bp. Conclusions Human single chain antibody to hepatitis C core antigen has been identified by means of the phage display technology.

关 键 词：噬菌体 丙型肝炎病毒 病毒核心蛋白 单链可变区体抗体 

分 类 号：R512.63[医药卫生—内科学] R373.21[医药卫生—临床医学]