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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 ,并获得了可溶性单链抗体的表达
Objective To screen and identify the coding sequence of human single chain variable region antibody (ScFv) against hepatitis B virus surface antigen (HBsAg) protein and to study the expression of small molecule single chain antibody in cells and the anti-HBV gene Treatment research to lay the foundation. Methods The phage display technique was used to purify the HBsAg protein by CsCl ultracentrifugation as solid phase antigen. After 5 cycles of “adsorption and elution amplification”, the phage single chain variable region antibody synthesis bank was used to obtain Positive HBsAg human single chain variable region antibody positive clones with strong antigen binding activity were obtained and their immunological detection and sequencing were performed. Results The ScFv fragment was 789 nt in length and encoded by 622 amino acid residues. It had typical structural characteristics of light chain and heavy chain variable regions and specificity of binding to HBsAg. Conclusion The phage antibody library technology was used to successfully obtain the HBsAg human single-chain variable region antibody gene and to obtain the soluble single-chain antibody