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目的利用噬菌体肽库技术筛选可特异性结合IL-2Rα的短肽序列,探索获得IL-2Rα小分子拮抗剂的可能性。方法以高表达IL-2Rα的MT-2细胞为钓饵,筛选噬菌体线性12肽库,用抗IL-2Rα单克隆抗体竞争洗脱结合的噬菌体,细胞ELISA、免疫组化鉴定阳性噬菌体克隆,并测序。结果随机挑选的17个克隆中有7个可与MT-2细胞结合。免疫组化显示阳性噬菌体克隆M15可与MT-2细胞及PHA刺激的PBMC结合。根据阳性克隆DNA序列,推导出氨基酸序列,共有6种序列,富含亲水氨基酸并包含Tyr、Phe、Leu保守残基。结论得到含Tyr、Phe保守残基的序列,阳性序列可结合细胞表面的IL-2Rα。
OBJECTIVE: To screen short peptide sequences that can specifically bind to IL-2Rα by phage peptide library technology and to explore the possibility of obtaining IL-2Rα small molecule antagonists. Methods MT-2 cells with high expression of IL-2Rα were used as bait to screen phage linear 12-peptide library. Anti-IL-2Rα monoclonal antibody was used to compete for elution of bound phage. Cell ELISA was used to identify positive phage clones by immunohistochemistry. . Results Seven out of 17 randomly selected clones were able to bind to MT-2 cells. Immunohistochemistry showed that positive phage clone M15 could bind to MT-2 cells and PHA-stimulated PBMCs. Based on the positive cloned DNA sequences, the deduced amino acid sequence has 6 sequences, which are rich in hydrophilic amino acids and contain Tyr, Phe and Leu conserved residues. Conclusion The sequence containing Tyr and Phe conserved residues was obtained. The positive sequence could bind to IL-2Rα on cell surface.