人源抗丙型肝炎病毒包膜蛋白E2单链抗体的研究

目的 研制抗丙型肝炎病毒包膜蛋白E2（HCVE2）的人源噬菌体单链抗体，并探讨其临床价值。方法 以重组的HCVE2为固相抗原，利用亲和筛选的原理，从噬菌体抗体库中经过5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HCVE2的人源单链抗体；用该抗体对10例石蜡包埋的丙型肝炎患者肝组织进行免疫组织化学鉴定。结果 ELISA结果表明，制备的HCVE2人源单链抗体（吸光度值A450为1.88)能与HCVE2抗原特异性结合，免疫组织化学结果表明，该抗体能够特异性识别丙型肝炎患者肝组织HCVE2抗原，与正常肝组织及乙型肝炎病毒（HBV）抗原均无交叉反应。结论 此法制备的单链抗体亲和性好，特异性强，且制备方法简便，周期短，为HCVE2病原的检测提供了新的有效的试剂。

Preparation of human single chain Fv antibody specifically against hepatitis C virus E2 antigen

Objective To identify human single chain Fv antibody (ScFv) against hepatitis C viral E2 antigen and its value clinically. Methods The recombinant phages were panned by E2 antigen which was coated in a microtiter plate. After five rounds of biopanning, 56 phage clones were identified specific to E2 antigen. The affinity and specificity of ScFv were evaluated by ELISA and immunohistochemistry, respectively. Results The data of E2-ScFV DNA digestion and DNA sequencing showed that the ScFv gene was composed of 750bp. ELISA and immunohistochemistry demonstrated that the human single chain Fv antibody against HCV E2 antigen had a specific combination character with hepatitis C virus E2 antigen. Conclusions ScFv, having a sutestantial affnity and specificity and being easy to prepare, is valutole in the detec- tion of HCV E2 antigen.