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  

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

作　　者：郭忠[1] 张清春[1] 雷政登[1] 孔亮[1] 毛希琴[1] 邹汉法[1] 

机构地区：[1]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

出　　处：《高等学校化学学报》2002年第7期1277-1280,共4页Chemical Journal of Chinese Universities

基　　金：国家杰出青年基金 (批准号 :2 972 5 5 12 )

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 1 0 - 1 3mol,甚至几个 1 0 - 1Peptide 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.

关 键 词：固定化酶 纳升微反应器 蛋白质 纳升反应器 肽谱分析 痕量分析 毛细管微柱 

分 类 号：Q51[生物学—生物化学] Q503