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

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可变区抗体的编码基因。

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

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.