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

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 - 1

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.