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目的 筛选乙型肝炎病毒 (HBV)表面抗原 (HBsAg)的人源噬菌体单链抗体 ,并探讨其在临床治疗和诊断中的应用价值。方法 以HBsAg阳性血清超速离心纯化的HBsAg为固相抗原 ,从噬菌体单链可变区半合成抗体库中经过 5轮“吸附 洗脱 扩增”筛选过程 ,获得特异性较强的HBsAg人源单链可变区抗体 (ScFv) ;用该抗体对 10例石蜡包埋的乙型肝炎患者肝组织进行免疫组化鉴定。结果 酶联免疫吸附法 (ELISA)结果表明 ,制备的HBV人源单链抗体能与HBsAg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乙型肝炎患者肝组织中的HBsAg抗原 ,与正常肝组织及丙型肝炎病毒 (HCV)的抗原均无交叉反应。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HBV病原的检测提供了新的有效试剂 ,为今后HBsAg人源抗体的研究和应用奠定了基础。
Objective To screen human phage scFv against hepatitis B virus surface antigen (HBsAg) and to explore its value in the clinical treatment and diagnosis. Methods HBsAg-positive serum was purified by ultracentrifugation using HBsAg as solid-phase antigen. After 5 rounds of “elution-elution amplification” screening from the semi-synthetic phage antibody library of phage single chain variable region, Chain variable region antibody (ScFv). The antibody was used to identify the liver tissues of 10 patients with paraffin-embedded hepatitis B by immunohistochemistry. Results The results of enzyme-linked immunosorbent assay (ELISA) showed that the prepared HBV human single chain antibody could specifically bind to HBsAg antigen. The results of immunohistochemistry showed that the antibody could specifically recognize the HBsAg antigen in the liver tissue of patients with hepatitis B , No cross-reaction with the normal liver and hepatitis C virus (HCV) antigens. Conclusion The single-chain antibody prepared by this method has good affinity and strong specificity, and the preparation method is simple and short, which provides a new effective reagent for the detection of HBV pathogens, which lays the foundation for the future research and application of HBsAg human antibodies .