胰岛素受体短肽配体的亲和筛选

Screening of Short Peptides Binding Specifically to Insulin Receptor

以高效表达胰岛素受体的中国仓鼠卵巢细胞(CHO-IR)为筛选靶标,经过一轮酸洗脱和三轮特异性竞争洗脱筛选,成功地富集了与胰岛素受体结合的重组噬菌体克隆.随机挑取噬菌体克隆检测其与胰岛素受体的结合活性,选取结合力最强的6个单克隆,测得其与CHO-IR之间的Kd值为纳摩尔级,表明为特异性结合;经序列分析表明短肽之间有明显的基序存在.

We used Chinese Hamster Ovary Cells over expressing insulin receptors (CHO IR) as screening targets to select peptide ligands of insulin receptors from a phage display random hexapeptide library. After one round of acid elution and three rounds of competitive elution, recombinant phage clones were enriched, the specific binding affinities of which were assayed with the method of cell ELISA and the K d values of the six most positive clones among them were calculated. Furthermore, we found several motifs from the sequences of these six clones. The result is very valuable for studying on the interaction between insulin and insulin receptor and designing small molecular drugs.