摘要
目的 筛选、鉴定抗乙型肝炎病毒 (HBV)表面抗原 (HBsAg)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 ,以氯化铯超速离心法纯化的HBsAg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”淘洗过程 ,获得抗原结合活性较强的HBsAg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 789nt,编码的产物由 2 62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HBsAg结合的特异性。结论 利用噬菌体抗体库技术 ,成功地获得了HBsAg人源单链可变区抗体的编码基因 。
Objective Screening and characterization of human phage antibody to hepatitis B virus surface antigen (HBsAg).Methods The recombinant phages were panned by HBsAg antigen which was coated in a microtiter plate, after five rounds of biopanning, 30 clones were demonstrated specific to HBsAg antigen. The specificity of ScFv had been demonstrated by ELISA.Results HBsAg phage antibody had a specific combination capacity with HBsAg antigen. The DNA sequence data showed that the ScFv gene of HBsAg was composed of 789 bp.Conclusion Human single chain antibody to HBsAg antigen has been identified by means of the phage display technology.
出处
《肝脏》
2000年第3期130-132,共3页
Chinese Hepatology
