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目的 研制抗丙型肝炎病毒包膜蛋白Ｅ２（ＨＣＶ Ｅ２）的人源噬菌体单链抗体，并探讨其临床价值。方法 以重组的ＨＣＶ Ｅ２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ＨＣＶ Ｅ２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ＨＣＶ Ｅ２人源单链抗体（吸光度值Ａ４５０为 １．８８）能与ＨＣＶ Ｅ２抗原特异性结台；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ＨＣＶ Ｅ２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ＨＣＶ Ｅ２病原的检测提供了新的有效的试剂。
Objective To develop human phage scFv against hepatitis C virus envelope protein E2 (HCV E2) and explore its clinical value. Methods The recombinant HCV E2 was used as solid phase antigen. After 5 rounds of “adsorption-elution-amplification” screening process and enzyme-linked immunosorbent assay (ELISA) and DNA sequencing from phage antibody library by affinity screening, Analysis of the obtained human E2 antibody single chain antibody; with the antibody in 10 cases of paraffin-embedded hepatitis C patients with liver tissue immunohistochemistry. Results The results of ELISA showed that the HCV E2 humanized single chain antibody (absorbance A450 was 1.88) could specifically bind to HCV E2 antigen. The results of immunohistochemistry showed that this antibody could specifically recognize hepatitis C Tissue HCV E2 antigen, no cross-reaction with normal liver and hepatitis B virus (HBV) antigen. Conclusion The single-chain antibody prepared by this method has good affinity and specificity, and the preparation method is simple and short cycle, which provides a new effective reagent for the detection of HCV E2 pathogen.