固定化酶纳升微反应器用于痕量蛋白质快速肽谱分析的研究

高等学校化学学报 ›› 2002, Vol. 23 ›› Issue (7): 1277.

固定化酶纳升微反应器用于痕量蛋白质快速肽谱分析的研究

郭忠, 张清春, 雷政登, 孔亮, 毛希琴, 邹汉法

中国科学院大连化学物理研究所, 国家色谱研究分析中心, 大连 116011

收稿日期:2001-04-29 出版日期:2002-07-24 发布日期:2002-07-24
通讯作者: 邹汉法(1961年出生),男,博士,博士生导师,主要从事基体辅助激光解吸离子化飞行时间质谱、生物液相色谱及毛细管电色谱研究.
基金资助:
国家杰出青年基金(批准号:29725512)

Studies on Rapid Micro-scale Peptide Mapping Analysis Using a Capillary Micro-reactor

GUO Zhong, ZHANG Qing-Chun, LEI Zheng-Deng, KONG Liang, MAO Xi-Qin, ZOU Han-Fa

National Chromatographic R & A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116011, China

Received:2001-04-29 Online:2002-07-24 Published:2002-07-24

摘要/Abstract

摘要： 利用毛细管作为酶固定化的载体,将酶直接键合到毛细管内壁,制成毛细管纳升反应器,结合质谱分析水解产物,获得了蛋白质的肽谱.实验发现,以毛细管为反应器后,蛋白质肽谱分析所需量大大减少,只需10^-13mol,甚至几个10^-15mol的量就可满足分析要求.

关键词: 纳升反应器, 毛细管固定化酶, MALDI-TOF-MS, 肽谱分析

Abstract: Peptide mapping analysis by combining the enzymatic digestion and the MALDI-TOF-MS has emerged as a powerful tool for identification and characterization of protein. We describe a nanoliter micro-reactor prepared with immobilization of enzyme on the porous wall of a capillary. The advantages of the prepared micro-reactor for digestion of the proteins are the elimination of enzyme autolysis and concentration of the micro sample. Horse heart cytochrome c is selected as the target protein for the evaluation of micro reactor, and it is found that about 0.1 picomole of protein analyte could routinely be digested for peptide mapping analysis and even femtomole of protein can be well characterized.

Key words: Nano liter micro reactor, Capillary immobilized enzyme, MALDI TOF MS, Peptide mapping analysis

中图分类号:

O657

TrendMD:

郭忠, 张清春, 雷政登, 孔亮, 毛希琴, 邹汉法. 固定化酶纳升微反应器用于痕量蛋白质快速肽谱分析的研究. 高等学校化学学报, 2002, 23(7): 1277.

GUO Zhong, ZHANG Qing-Chun, LEI Zheng-Deng, KONG Liang, MAO Xi-Qin, ZOU Han-Fa . Studies on Rapid Micro-scale Peptide Mapping Analysis Using a Capillary Micro-reactor. Chem. J. Chinese Universities, 2002, 23(7): 1277.