CPSA Shanghai 2011
Changing Paradigm in Drug Discovery & Development:
East Meets West
April 13 - 16, 2011
Renaissance Shanghai Pudong Hotel
Shanghai, China
Abstract
Friday AM - Plenary
Multi-dimensional Chromatography-Mass Spectrometry for Protein Analysis
Xiangmin Zhang
Department of Chemistry & Institutes of Biomedical Sciences
Fudan University, Shanghai 200433, China.
Email: xmzhang@fudan.edu.cn
Top-down strategy based on intact protein separation by multidimensional chromatography followed by mass spectrometric identification is proven to be more advantageous over bottom-up proteomic analytical strategies. Intact protein contains information of post-translational modifications, quantitative concentration, and so on. The information is of a great significance for understanding protein's functions, expressional level, disease diagnosis, etc. In this study, multidimensional liquid chromatography (MDLC) systems with array-column based reversed-phase separations were demonstrated. Methods to high-efficiency separation of intact protein were optimized. High abundance protein depletion using MDLC system was also discussed. Technologies for fast protein digestion of MDLC fractions were developed and applied to protein identification by mass spectrometry. A human liver proteome sample was analyzed for verifying running efficiency of the systems. Over 210 high abundance proteins were depleted by fractionations. Around 2500 proteins were identified after separation, fast on-line/ on-target-plate digestion, and MS analysis for a typical run. Hundreds of proteins with variety of post-translational modification were also identified. (This work is supported by the National Science Foundation of China, project 21027002, and the 973 project 2007CB914100.)