论文部分内容阅读
目的筛选、鉴定抗丙型肝炎病毒 (HCV)非结构蛋白 NS3的人单链可变区抗体 (Sc Fv) ,以解决人体内应用鼠单抗时的免疫原性问题 ,为进行抗 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 HCV非结构蛋白 NS3为固相抗原 ,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人单链可变区抗体的阳性克隆 ,并对其进行免疫检测及序列测定。结果筛选出来的 Sc Fv片段具有抗 NS3的特异性。证实利用噬菌体抗体库技术 ,可以成功地获得 HCV NS3人单链抗体 Sc Fv的编码基因。结论筛选获得了 HCV非结构蛋白 NS3的特异性单链抗体的编码基因。
Objective To screen and identify the human single chain variable region antibody (Sc Fv) against the nonstructural protein NS3 of hepatitis C virus (HCV) in order to solve the immunogenicity problem of human monoclonal antibody (McAb) Treatment research opens up new avenues. Methods The phage display technique was used to detect the antigen binding activity after 5 rounds of “adsorption-elution-amplification” screening process from the phage single chain variable region antibody library using the recombinant HCV NS3 as solid phase antigen Strong positive clone of HCV NS3 human single chain variable region antibody, and its immune detection and sequencing. Results Sc Fv fragments were screened for anti-NS3 specificity. Confirmed that the use of phage antibody library technology can be successfully obtained HCV NS3 human scFv single chain antibody encoding genes. Conclusion The gene encoding specific single chain antibody of NS3, a nonstructural protein of HCV, was screened.