胰岛素受体短肽配体的亲和筛选

胰岛素受体短肽配体的亲和筛选

朴锦华, 李鸿梅, 王丽萍，高利增, 石玉华, 李惟

吉林大学生命科学学院，长春 130023

收稿日期:2003-05-05 修回日期:1900-01-01 出版日期:2003-07-26 发布日期:2003-07-26
通讯作者: 李惟

Screening of Short Peptides Binding Specifically to Insulin Receptor

PIAO Jin-hua, LI Hong-mei, WANG Li-ping, GAO Li-zeng, SHI Yu-hua, LI Wei

College of Life Science, Jilin University, Changchun 130021, China

Received:2003-05-05 Revised:1900-01-01 Online:2003-07-26 Published:2003-07-26
Contact: LI Wei

摘要/Abstract

摘要： 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Ｋ_d值为纳摩级，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.

关键词: 胰岛素受体, 噬菌体展示随机六肽库, Ｋd值

Abstract: 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 thes esix clones. The result is very valuable for studying on the interaction betwee n insulin and insulin receptor and designing small molecular drugs.

Key words: insulin receptor, phage display random hexapeptide library, K_d value

中图分类号:

Q513.2

朴锦华, 李鸿梅, 王丽萍，高利增, 石玉华, 李惟. 胰岛素受体短肽配体的亲和筛选[J]. J4, 2003, 41(03): 384-387.

PIAO Jin-hua, LI Hong-mei, WANG Li-ping, GAO Li-zeng, SHI Yu-hua, LI Wei. Screening of Short Peptides Binding Specifically to Insulin Receptor[J]. J4, 2003, 41(03): 384-387.