物理化学学报 >> 2005, Vol. 21 >> Issue (05): 508-511.doi: 10.3866/PKU.WHXB20050510

研究论文 上一篇    下一篇

利用pⅧ展示系统改进噬菌体抗体芯片

廖玮; 洪龙; 魏芳; 朱圣庚; 赵新生   

  1. 北京大学化学与分子工程学院化学生物学系, 分子动态与稳态结构国家重点实验室; 北京大学生命科学学院, 北京    100871
  • 收稿日期:2004-10-29 修回日期:2004-12-10 发布日期:2005-05-15
  • 通讯作者: 赵新生 E-mail:zhaoxs@pku.edu.cn

Improving Phage Antibody Chip by pⅧ Display System

LIAO Wei; HONG Long; WEI Fang; ZHU Sheng-Geng; ZHAO Xin-Sheng   

  1. State Key Laboratory of Molecular Dynamic and Stable Structures, Department of Chemical Biology, College of Chemistry and Chemical Engineering, Peking University;    College of Life Sciences, Peking University, Beijing    100871
  • Received:2004-10-29 Revised:2004-12-10 Published:2005-05-15
  • Contact: ZHAO Xin-Sheng E-mail:zhaoxs@pku.edu.cn

摘要: 将展示单链抗体的重组噬菌体与羧基终止的硅片偶联, 制成噬菌体抗体芯片, 可用于检测多类蛋白质和蛋白质组. 通常抗体被展示于噬菌体外壳蛋白pⅢ上, 由此制备的芯片灵敏度和信噪比较低. 我们选用凝血酶特异的单链抗体为代表, 比较了pⅢ展示系统和pⅧ展示系统制成芯片的检测效果. 由于pⅧ展示系统的融合蛋白拷贝数多, 所受空间位阻小, 大幅度提高了噬菌体抗体芯片的灵敏度和信噪比, 有望用于制备新型蛋白质芯片.

关键词: 蛋白质芯片, 噬菌体展示抗体, pⅢ展示系统, pⅧ展示系统, 凝血酶特异的单链抗体

Abstract: Phage antibody chips, prepared by coupling the recombinant phage displaying scFv with carboxyl-terminated silicon, can be applied in detecting proteins and proteomes. In previous work, we immobilized phages displaying scFv on the coat protein pⅢ on silicon surface as model chips. To improve the sensitivity and the ratio of signal to noise, in this work, scFv specific binding to thrombin was displayed by pⅧ display system, which is the major coat protein on the phage. As the results of changing from pⅢ to pⅧ display, the sensitivity and the ratio of signal to noise of chips were enhanced prominently, due to the increase of the copy number and the decrease of steric hindrance. It is concluded that scFv fused to the coat protein pⅧ is a promising candidate for the preparation of novel protein chips.

Key words: Protein chips, Phage-display antibody, pⅢ display system, pⅧ display system, scFv specific to thrombin