从噬菌体肽库中筛选并鉴定中期因子特异性结合肽  

Screening and identification of peptides that bind human midkine selected from phage display library

作　　者：沈建芬[1] 闵丽姗 姚行[3] 钱福初 戴利成 

机构地区：[1]嘉兴市第一医院中心实验室,浙江314000 [2]湖州市分子医学重点实验室 [3]湖州市中心医院肝胆外科

出　　处：《中华实验外科杂志》2013年第6期1278-1280,共3页Chinese Journal of Experimental Surgery

基　　金：湖州市科技局重大科技攻关计划资助项目（2005GS01）

摘　　要：目的筛选并鉴定可与中期因子（MK）特异性结合的短肽。方法以人重组MK蛋白为靶分子，分别从噬菌体随机7肽和环7肽展示库中筛选出与MK蛋白高特异性结合的阳性噬菌体克隆。酶联免疫吸附试验（ELISA）鉴定所挑选噬菌体和MK蛋白的亲和力。噻唑蓝（MTT）比色法体外评价该合成短肽对HepG2细胞生长的抑制作用。结果经过3轮淘洗，筛选出的20个阳性克隆与MK蛋白均具有一定结合力，测序并推导出优选的3条线性肽和3条环肽的氨基酸序列，合成肽能不同程度抑制HepG2细胞的增殖，其中MK-P3（CTSTAMDNC）抑制HepG2增殖作用最强（P〈0．01），其对HepG2肝癌细胞的抑制率可达40．7％。结论成功筛选出了4条可和MK蛋白结合并对肿瘤细胞有一定抑制作用的短肽。Objective To screen human midkine （h-midkine） inhibitory peptides from the phage display peptide library. Methods Recombinant human midkine （rh-midkine） protein was used to search for the specific peptides by panning. The specific binding activities of positive clones to target protein were examined by phage enzyme linked immunosorbent assay （ELISA） . The effects of binding peptides on HepG2 cells proliferation were analyzed by methyl thiazol tetrazolium （MTT） method. Results Thenty positive clones were obtained after three rounds of selection. Three linear peptides and three cycle pep- tides, more frequently occurring in the affinity-selected phage population and higher specificity to target protein, were chemically synthesized. The synthetic peptides produced inhibition of the HepG2 cells prolif- eration to different degrees, the MP-3 （CTSTAMDNC） has the strongest inhibitory effect for the HepG2 proliferation （P 〈0. 01 ）, the inhibition rate is about 40. 7%. Conclusion Four peptides that can specifically bind to MK protein have been screened successfully, and these four peptides show suppression effects on the proliferation in tumor cells.

关 键 词：中期因子 噬菌体展示肽库 肽 

分 类 号：R[医药卫生]