Wang Lei, Qiao Yuan-yuan, Wang Gang, Chen Yu-ping, Yang Jun, Wang Yan, Liu Yu-feng
Department of Dermatology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2005 Mar;21(2):185-7.
To screen human scFv against IL-8 from phage antibody library.
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.
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 Vlambda gene belonged to human VlambdaDPL5 and VlambdaDPL2 subgroups,respectively.
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.
从噬菌体抗体库中筛选抗白细胞介素-8（IL-8）的人源单链抗体片段（scFv）。
将原核表达载体pRSET-IL-8转染至大肠杆菌BL21（DE3）中表达IL-8-组氨酸（His）融合蛋白，并用亲和层析法进行纯化。用该融合蛋白对噬菌体抗体库进行3轮淘选，筛选特异性抗体。测定并分析阳性克隆的抗原结合活性及DNA序列。
经过3轮淘选，获得2个能特异性结合IL-8的阳性克隆。DNA序列分析显示，2个重链可变区（VH）基因均属于人IgG VH3亚组，轻链可变区（Vlambda）基因分别属于人VlambdaDPL5和VlambdaDPL2亚组。
利用噬菌体展示技术获得人抗IL-8抗体是可行的，为进一步研究银屑病及其他相关疾病奠定了基础。
