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目的: 从噬菌体抗体库中筛选人源性抗IL- 8单链抗体(scFV)。方法: 采用原核表达载体pRSET- IL- 8在大肠杆菌BL21(DE3)中表达IL- 8 His融合蛋白, 并通过亲和层析纯化。以该融合蛋白为固相抗原, 对噬菌体抗体库进行 3轮淘筛, 并对所获阳性克隆进行抗原结合活性测定和DNA序列测定。结果: 获得 2株特异性抗IL- 8噬菌体抗体。对其DNA序列测定结果表明, 其VH基因均属于人IgGVH3亚群, Vλ基因分别属于人VλDPL5和VλDPL2亚群。结论: 利用噬菌体抗体库技术可不经免疫制备人源性抗IL- 8抗体, 为银屑病及相关疾病的临床应用提供了条件。
OBJECTIVE: To screen human anti-IL-8 single chain antibody (scFV) from phage antibody library. Methods: The prokaryotic expression vector pRSET-IL-8 was used to express IL-8 His fusion protein in E. coli BL21 (DE3) and purified by affinity chromatography. Using the fusion protein as solid phase antigen, the phage antibody library was panned for 3 rounds and the positive clones were tested for antigen binding activity and DNA sequencing. Results: Two specific anti-IL-8 phage antibodies were obtained. The results of DNA sequencing showed that the VH genes belong to the human IgGVH3 subgroup, and the Vλ genes belong to the human VλDPL5 and VλDPL2 subgroups. Conclusion: The phage antibody library can be used to prepare human anti-IL-8 antibody without immunization, which provides the conditions for the clinical application of psoriasis and related diseases.