抗丙型肝炎病毒核心蛋白单链抗体的制备及免疫组织化学研究  

Preparation and immunohistochemical study of human single chain Fv antibody against hepatitis C virus core antigen

作　　者：钟彦伟[1] 成军[1] 焦成松[2] 张忠东[1] 李强[1] 李莉[1] 陈菊梅[1] 

机构地区：[1]解放军第302医院传染病研究所基因治疗研究中心,北京100039 [2]广州军区武汉总医院

出　　处：《解放军医学杂志》2004年第1期8-9,12,共3页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金资助课题 (编号 3990 0 1 30 )

摘　　要：目的　制备丙型肝炎病毒核心蛋白单链抗体并进行免疫组织化学研究。方法　以大肠杆菌表达的重组丙型肝炎病毒(HCV)核心蛋白为固相抗原 ,利用抗原 抗体亲和性结合的原理 ,从半合成的人源可变区噬菌体抗体库中经过 3轮“吸附 洗脱 扩增”的筛选过程及酶联免疫吸附试验 (ELISA)和DNA序列分析 ,获得HCV核心蛋白的人源单链抗体 ;用该抗体对 772 1转染全长HCVcDNA后筛选的阳性克隆细胞中的HCV核心蛋白抗原进行免疫组化鉴定。结果　ELISA结果表明 ,制备的HCV核心蛋白人源单链抗体能与HCV核心蛋白抗原特异性结合 ;免疫组化结果表明 ,该抗体能够特异性识别HCV核心蛋白抗原 ,与正常肝细胞无交叉反应。结论　此法制备单链抗体方法简便 ,周期短 ,可为HCV核心蛋白病原的检测提供新的检测手段。Objective To detect hepatitis C virus core antigen in 7721 cells transfected with HCV cDNA by immunohistochemistry method with human single chain Fv antibody(scFv). Methods The recombinant phages were panned by core antigen which was coated in a microtiter plate. After three rounds of biopanning, 48 clones were identified specific to core antigen. The affinity and specificity of scFv were evaluated by ELISA and immunohistochemistry. Results ScFv-core DNA digestion and sequence data showed that the scFv gene was composed of 774bp. Conclusion Human single chain Fv antibody against HCV core antigen has a specific combining capacity with hepatitis C virus core antigen.

关 键 词：C型肝炎样病毒属 病毒核心蛋白质类 单链抗体 免疫组织化学 

分 类 号：R512.63[医药卫生—内科学]