全自動多次元タンパク質プロファイリングシステムを用いるヒト血漿プロテオーム解析

抄録

Blood plasma is not only the primary clinical specimen but also represents that the largest and deepest version of human proteome present in any sample. In clinical and diagnostic proteomics for human plasma, it is essential to develop a comprehensive and robust system for a large-scale proteome analysis. Recently, multi-dimensional liquid chromatography/ tandem mass spectrometry (LC/MS/MS) systems have been developed as powerful tools especially for protein identification of highly complexes and have achieved resolution power compatible to two-dimensional gel electrophoresis. However, these systems need to be further developed in terms of ease in usage and industrial applicability to the routine clinical use, because their application to clinical researches requires an analysis of a large number of human samples with stability and reproducibility. Therefore, we have constructed a technically simplified and high-speed/throughput LC/MS/MS system with micro-flowing reversed-phased chromatography and linear ion-trap mass spectrometry equipped with nanoelectrospray interface. Additionally, the system was combined with on-line strong cation exchange chromatography as a multi-dimensional protein profiling system. For a large-scale human plasma proteome analysis, we have also established protein depletion, in-solution digestion and data-integrating/mining systems with an automated operation. Here, we will present the application of our fully automated multi-dimensional protein profiling system to plasma proteome analysis for clinical proteomics.