从噬菌体抗体库筛选人源性抗白细胞介素8抗体

Screening of human scFv against IL-8 from phage antibody library

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。

AIM: To screen human scFv against IL-8 from phage antibody library. METHODS: IL-8-His fusion protein was expressed in E.coli BL21(DE3) transfected with prokaryotic expression vector pRSET-IL-8 and was purified by affinity chromatography. Specific antibody was screened by 3 rounds of panning of phage antibody library with the fusion protein. The antigen binding activity and DNA sequences of positive clones were determined and analyzed. RESULTS: After 3 rounds of panning, 2 positive clones were obtained which could bind IL-8 specifically. DNA sequence analysis showed both of the 2 VH genes belonged to human IgG VH3 subgroup, and the Vλ gene belonged to human VλDPL5 and VλDPL2 subgroups, respectively. CONCLUSION: It is feasible to obtain human anti-IL-8 antibody by phage display technique, which provides the basis for further research on psoriasis and other related diseases.