HCV非结构蛋白NS5A人源单链可变区抗体基因的筛选与鉴定

目的 筛选、鉴定抗丙型肝炎病毒（HCV）非结构蛋白NS5A的人源单链可变区抗体（ScFv)的编码基因，为HCV NS5A蛋白质生物学功能的研究及抗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，以重组纯化的HCV非结构蛋白NS5A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”筛选过程，获得抗原结合活性和特异性较强的HCVNS5A人源单链可变区抗体基因片段的阳性克隆，并对其进行免疫检测及序列测定，结果 筛选得到的ScFv片段基因核苷酸序列为789nt，编码由262个氨基酸残基组成的多肽，具有典型的免疫球蛋白轻链和重链可变区的结构特点，基因编码产物具有HCV NS5A蛋白反应的免疫学活性和特异性。结论 利用噬菌体抗体库技术，成功获得了HCV NS5A人单链可变区抗本ScFv编码基因，并获得了可溶性单链抗体的表达。

Screening and characterization of human phage antibody to hepatitis virus C NS5A antigen

Objective To screen and characterize human phage antibody against hepatitis C virus (HCV)NS5A antigen.Methods The recombinant phages were panned by recombinant HCV NS5A antigen which was coated in a microtiter plate;After five rounds of biopanning,35 clones were obtained and demonstrated specific to NS5A antigen.The specific binding capacity of the ScFv antibody to HCV NS5A antigen was determined by ELISA.Results HCV NS5A phage antibody had a specific combination capacity with hepatitis C virus NS5A antigen.The DNA sequence data showed that the ScFv gene was composed of 789 bp and codes for a peptide of 262 amino acid residues.Conclusion Human single chain antibody to hepatitis C virus NS5A antigen has been identified by means of the phage display technology.