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目的 筛选、鉴定抗丙型肝炎病毒 (HCV)非结构蛋白 (NS) 3的人单链抗体 (ScFv) ,以解决鼠单抗的免疫原性问题 ,为HCV的基因治疗研究开辟新途径。方法 采用噬菌体表面展示技术 ,以重组的HCVNS3为包被抗原 ,从噬菌体单链可变区抗体库中经过 5轮“吸附 洗脱 扩增”筛选过程 ,获得抗原结合活性较强的HCVNS3人单链抗体 (ScFv片段 )阳性克隆 ,并对其进行免疫及序列测定。结果 筛选出来的ScFv片段具有抗NS3的特异性。结论 利用噬菌体抗体库技术 ,成功获得HCVNS3人单链抗体ScFv片段
Objective To screen and identify human single chain antibody (ScFv) against hepatitis C virus (HCV) nonstructural protein (NS) 3 in order to solve the immunogenicity of mouse monoclonal antibody and to explore a new way for gene therapy of HCV. Methods Phage display technique was used to screen HCV NS3 antigen as coat antigen. After 5 rounds of “elution-elution amplification” screening from the phage single-chain variable region antibody library, HCV NS3 human single-stranded Antibodies (ScFv fragment) were cloned and immunized and sequenced. Results ScFv fragments screened for anti-NS3 specificity. Conclusion The phage antibody library technology successfully obtained HCV NS3 human single chain antibody ScFv fragment