应用噬菌体随机肽库技术研究干扰素-α2b抗原表位  

Epitope mapping of interferon-α2b using phage random peptide library

作　　者：田旺[1] 李正赫[1] 曹学琳 杨文博[1] 白钢[1] 

机构地区：[1]南开大学生命科学学院 [2]天津华利达生物工程有限公司

出　　处：《中华微生物学和免疫学杂志》2005年第7期578-582,共5页Chinese Journal of Microbiology and Immunology

基　　金：天津市科技攻关重点项目(033104611)

摘　　要：目的应用噬菌体随机肽库技术,研究干扰素-α2b高抗原性表位。方法利用结合有干扰素-α2b多克隆抗体的免疫磁性微球,对噬菌体随机肽库进行生物淘筛,以ELISA方法鉴定阳性克隆。阳性克隆测序后与干扰素氨基酸序列进行了同源性分析,研究干扰素-α2b分子中高抗原性表位。结果经4轮淘筛后噬菌体克隆的阳性率为57.6%,据随机挑选的12个阳性克隆的同源性分析结果将其分为3组,分别对应干扰素-α2b3个高抗原性表位与预测的结果吻合,其中第2组与干扰素受体结合区AB环接近。结论利用噬菌体随机肽库技术,成功筛选出3个与抗原性预测相符合的干扰素-α2b抗原表位,为研究干扰素分子的作用机理、制备抗特定表位的单克隆抗体以及设计和开发干扰素的小分子模拟肽奠定了基础。Objective To identify the epitope with strong antigenicity of IFN-α2b by phage random peptide library. Methods The polyclonal antibodies against IFN-α2b binding to immunomagnetic microspheres was chosen as target protein, the biopanning of phage random peptide library was carried out and the positive clones was characterized by ELISA. After homologous analysis of amino acid sequence of positive clones with IFN-α2b, the epitope with strong antigenicity of IFN-α2b was studied. Results After 4 rounds of effective screening, the positive rate was 57.6% （53/92）. Twelve positive clones were sequenced and homologous analysis of amino acid sequences with IFN-α2b showed that 3 groups were obtained. The sequences of group Ⅱ exhibited structural homology with Loop AB（29-35） of IFN-α2b. Condusion Three epitopes with strong antigenicity of IFN-α2b were obtained. The results can be applied to the research on functional mechanism of IFN-α2b and preparation of monoclonal antibody against the special epitope as well as the mimic minimolecular design and development of IFN-α2b.

关 键 词：噬菌体随机肽库 免疫磁性微球 抗原表位 干扰素-Α2B 噬菌体随机肽库技术 ELISA方法 多克隆抗体 同源性分析 阳性克隆 氨基酸序列 

分 类 号：R392[医药卫生—免疫学]