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目的利用噬菌体表面展示技 术,从半合成人源化单链可变区抗体库中筛选、鉴定丙型肝炎病毒(HCV)非结构蛋白NS4A 的单 链可变区抗体(ScFv)及其编码基因,为抗HCV的细胞内免疫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NS4A为包被抗原,从噬菌体单链可变区抗体库中经过3轮“吸附-洗脱-扩增”筛选过 程,获得抗原结合活性较强的HCV NS4A人单链可变区抗体片段阳性克隆,并对其 进行免疫学活性及编码基因序列测定。结果筛选得到 的ScFv片段具有抗HCV NS4A的特异性,编码基因符合人源化单链可变区抗体编码基因的特 点。结论利用噬菌体抗体库技术,成功获得HCV NS4A的特异性人抗体,为进一步研究HCV NS4A的生物学功能及细胞内免疫抗HCV基因治疗方 案奠定基础。
OBJECTIVE: To screen and identify single-chain variable region antibody (ScFv) of non-structural protein NS4A of hepatitis C virus (HCV) and its coding gene by semi-synthetic humanized single-chain variable region antibody library by phage display technology. To open up new avenues for anti-HCV intracellular immune gene therapy research. Methods Using phage display technique, the recombinant HCV NS4A was used as coating antigen. After three rounds of “adsorption-elution-amplification” screening process, the antigen binding activity was compared with that of phage single chain variable region antibody library Strong HCV NS4A human single chain variable region antibody fragment positive clones, and its immunological activity and coding gene sequence determination. Results The ScFv fragment was screened for its specificity against HCV NS4A. The coding gene was in line with the characteristics of the humanized single chain variable region antibody - encoding gene. Conclusion The phage antibody library technology was used to successfully obtain the specific human antibody against HCV NS4A, which lays the foundation for further study on the biological function of HCV NS4A and the intracellular immune anti-HCV gene therapy.