北京大学学报(自然科学版)

ICAM-1特异结合肽的淘汰与性质研究

赵乐群,赵佳慧,程庆砾,黄仪秀,朱圣庚   

  1. 北京大学生命科学学院,北京,100871;解放军总医院肾科,北京,100853
  • 收稿日期:2001-09-03 出版日期:2002-07-20 发布日期:2002-07-20

Selection of the Peptides Binding to ICAM-1 from Phage Display Random 15mer Peptide Library

ZHAO Lequn1,ZHAO Jiahui2,CHENG Qingli2,HUANG Yixiu1,ZHU Shenggeng1   

  1. 1College of Life Sciences, Peking University, Beijing, 100871, China; 2Department of Nephrology, General Hospital of PLA, Beijing, 100853, China
  • Received:2001-09-03 Online:2002-07-20 Published:2002-07-20

摘要: 利用噬菌体展示载体pCANTAB5G8构件了随机十五肽库,根据转化率计算库容量为1.36×108。用PCR-SSCP检验了库的异质性。以固相包被亲和淘选的方法,经过4轮淘选和ELISA分析,得到了5个可与重组人细胞间粘附分子1(ICAM-1)特异结合的噬菌体克隆,对所展示的十五肽进行了序列分析。其中活性较高的噬菌体展示十五肽与ICAM-1的亲和常数约为7.87×107

关键词: 噬菌体展示, 随机多肽库, 细胞间粘附分子1, 亲和淘选

Abstract: A phage display 15mer random peptide library expressed on the N-terminal of the filamentous phage M13 minor coat protein pⅢ was constructed using the phagmid pCANTAB5G8. The capacity of library is 1.36×108, calculated from the transformation efficiency. Human recombinant Intercellular Adhesion Molecule 1 was immobilized on the ELISA plates. After 4 rounds affinity panning and ELISA analysis, 5 positive phage clones that can bind to ICAM-1 specifically were selected. These positive clones were sequenced and the amino acid sequence of 15mer were deduced.The Ka of one positive clone was 7.87×107, measured by ELISA.

Key words: phage display, random peptide library, ICAM-1, biopanning

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