EpCAM分子特异结合多肽的筛选和鉴定  

The Selection and Characterization of the Peptide Binding to EpCAM

作　　者：贾晨霜 李欣瑶 陈利荣 袁悦 高倩 路征 徐婕 舒展 王怡文 侯颖春[1] JIA Chenshuang;LI Xinyao;CHEN Lirong;YUAN Yue;GAO Qian;LU Zheng;XU Jie;SHU Zhan;WANG Yiwen;HOU Yingchun(College of Life Sciences,Shaanxi Normal University,Xi'an 710119,China)

机构地区：[1]陕西师范大学生命科学学院,西安710119

出　　处：《中国细胞生物学学报》2021年第12期2326-2333,共8页Chinese Journal of Cell Biology

基　　金：中央高校基本科研业务费专项资金(批准号:2019TS076)资助的课题。

摘　　要：上皮/内皮细胞分子(EpCAM)是癌干细胞(CSC)重要的表面标记物分子。利用噬菌体展示肽库进行亲和消减富集生物淘筛(Biopanning),经过5轮淘筛,从最后一轮的噬菌体洗脱库中随机挑选47个克隆扩增,通过ELISA方法初步确定30个阳性噬菌体克隆并进行测序,序列分析后获得5个共有序列克隆群,随后通过细胞免疫荧光实验验证和鉴定了其CSC靶向结合特异性。经综合分析,确定R8噬菌体克隆的EpCAM结合特异性和敏感性最佳,将其多肽序列命名为ESP1。该EpCAM靶向肽ESP1未来在癌化疗靶向递送药物研发方面具有潜在使用价值,可使药物尽可能多地杀伤CSC,进而提高疗效,减少复发。EpCAM(epithelial/endothelial cell adhesion molecule)is an important surface marker of CSC(cancer stem cell).After 5 rounds of biopanning,47 clones were randomly selected from the last round of phage eluted library for amplification.Thirty positive phage clones were preliminarily identified by ELISA method,and sequentially sequenced.After sequence analysis,5 consensus sequence groups were concluded.Subsequently,several cultured cancer cells were tested and identified by immunofluorescence assay to evaluate its affinity to CSC.Based on the molecular and cellular test results,R8 clone presents the best specificity and sensitivity for EpCAM binding,and its peptide sequence was named as ESP1.ESP1 is of important potential to be used for the development of cancer targeted chemotherapy delivery in future,and such delivery will have CSCs killed as more as possible that improves the therapy efficacy and reduces recurrence.

关 键 词：上皮/内皮细胞分子 癌干细胞 靶向多肽 癌干细胞靶向化疗药物 

分 类 号：R965.1[医药卫生—药理学]