用2D Micro-HPLC/MS分析六种蛋白质混合物的胰蛋白酶解多肽产物

Analysis of Six Protein Mixture Digested by Trypsin Using 2D Micro-HPLC System with MS

采用2D Micro-HPLC系统,用MS对六种蛋白质的胰蛋白酶解多肽产物进行了分析,实现了蛋白质的高灵敏度、高分离度的检测。结合离子交换色谱法和反相毛细管色谱法,利用六个捕集柱和脱盐系统,可对各种分离条件进行优化。因此,可以实现对多种痕量酶解多肽的分离检测。该系统是痕量蛋白质和未知蛋白质检测的有力工具,结合高性能质谱仪,可以达到非常高的灵敏度。

Six protein mixtures digested were analyzed by trypsin using 2D Micro-HPLC system and MS. The high performance of 2D Micro-HPLC system can perform separation of complex peptide mixtures with a level of sensitivity and resolution that meets or exceeds present requirements. Combining cation exchange chromatography in the first dimension and capillary reversed-phase chromatography in the second dimension, and using 6 trap columns, allow for the various separation conditions to be optimized independently, which enables analysis for large numbers of trace peptides from digests. The proposed sys tern is an extremely effective tool for the discovery of unknown proteins with low expression levels. When this system is coupled with a high-performance mass spectrometer, unparalleled performance and sensitivity can be achieved.