ICAM-1特异结合肽的淘汰与性质研究

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

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

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/Abstract

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

中图分类号:

Q516

赵乐群,赵佳慧,程庆砾,黄仪秀,朱圣庚. ICAM-1特异结合肽的淘汰与性质研究[J]. 北京大学学报（自然科学版）.

ZHAO Lequn,ZHAO Jiahui,CHENG Qingli,HUANG Yixiu,ZHU Shenggeng. Selection of the Peptides Binding to ICAM-1 from Phage Display Random 15mer Peptide Library[J]. Acta Scientiarum Naturalium Universitatis Pekinensis.

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https://xbna.pku.edu.cn/CN/Y2002/V38/I4/474

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[2]	高新胜, 黄仪秀, 郭振泉, 朱圣庚, 陈玉珍. 小鼠噬菌体抗体库的构建和N-肽结合单链抗体的筛选[J]. 北京大学学报（自然科学版）, 2000, 36(4): 489-494.