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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 study the high antigenic epitope of interferon-α2b by phage display peptide library technology. Methods Immunomagnetic beads with polyclonal antibody against interferon α2b were used to screen the phage random peptide library. The positive clones were identified by ELISA. The positive clones were sequenced and homologous to the interferon amino acid sequence to study the high antigenic epitopes in the interferon-α2b molecule. Results The positive rate of phage clones after 4 rounds of panning was 57.6%. According to the results of homology analysis of 12 positive clones randomly selected, they were divided into 3 groups, corresponding to 3 high antigenic epitopes of interferon-α2b The predicted results are in agreement, with group 2 approaching the AB loop of the interferon receptor binding domain. Conclusions Three interferon-α2b epitopes were successfully screened by phage random peptide library technology. In order to study the mechanism of action of interferon molecules, preparation of monoclonal antibodies against specific epitopes and the design and development Interferon small molecule mimic peptides laid the foundation.