乙肝病毒核心抗原人源单链可变区抗体的筛选  

Screening on Human Phage Antibody of Hepatitis B Virus Core Antigen

作　　者：钟彦伟[1] 成军[1] 王刚[1] 田小军[1] 陈新华[1] 李莉[1] 陈菊梅[1] 张玲霞[1] 

机构地区：[1]中国人民解放军第三○二医院,北京100039

出　　处：《中国公共卫生》2002年第2期153-154,共2页Chinese Journal of Public Health

基　　金：国家自然科学基金资助 (3990 0 1 30 )

摘　　要：目的　筛选、鉴定乙型肝炎病毒 (HBV)核心抗原 (HBcAg)蛋白的人源单链可变区抗体 (ScFv)的编码基因 ,为细胞内表达小分子单链抗体的研究及抗HBV的基因治疗研究奠定基础。方法　采用噬菌体表面展示技术 ,以HBcAg蛋白为固相抗原 ,从噬菌体单链可变区抗体半合成库中经过 5轮“吸附 -洗脱 -扩增”筛选过程 ,获得抗原结合活性较强的HBcAg人源单链可变区抗体阳性克隆 ,并对其进行免疫检测及序列测定。结果　筛选得到的ScFv片段编码基因为 771nt,编码的产物由 2 5 7个氨基酸残基组成 ,具有典型的轻链和重链可变区结构特点以及与HBcAg结合的特异性。结论　利用噬菌体抗体库技术成功地获得了HBcAg人源单链可变区抗体的编码基因 ,并获得了可溶性单链抗体的表达。Objective To screen and characterize human-phage antibody of core antigen of hepatiti B virus (HBcAg).Methods Using HBc antigen as the coating antigen,the recombinant phage antibody to HBc antigen were panned from a semisynthetic phage library by phage display technique,after five rounds of biopanning,60 clones were demonstrated specific to HBcAg antigen.The specificity of ScFv was demonstrated by ELISA.Results HBcAg phage antibody has a specific combinating capacity with HBcAg antigen.The data DNA sequence showed that the ScFv gene of HBcAg was composed of 771 bp.Conclusion Human-single-chain antibody of HBcAg antigen was identified by means of the phage display technique.

关 键 词：噬菌体展示技术 乙型肝炎病毒 核心抗原 亲和筛选 噬菌体抗体 

分 类 号：R512.62[医药卫生—内科学]