DICP OpenIR
学科主题分析化学
New Separation and Identification Techniques for Proteome Analysis
Zhang YK(张玉奎)
会议文集36th International Symposium on High-Performance Liquid Phase Separations and Related Techniques
会议名称36th International Symposium on High-Performance Liquid Phase Separations and Related Techniques
会议日期2011-6-19
2011
会议地点布达佩斯
页码40-0
出版者待补充
出版地待补充
合作性质特邀报告
部门归属1810
主办者American Chemical Society
英文摘要The complexity of proteome brings great challenge and opportunity to promote the development of new techniques for liquid phase based proteome analysis. In our recent study, with newly developed methods and platforms, great improvement on proteome sample preparation, separation and identification was made. To improve the solubility of membrane proteome, and enzymatic biocompatibility with tryptic digestion, ionic liquids, BMIM BF4 showed superiority compared to the commonly applied methods, including methanol, urea and Rapist. Such a technique was successfully applied into the membrane proteome analysis of rat brain, and with the combination of 2D-HPLC-ESI-MS/MS, over 2000 non-abundant proteins, including 608 membrane proteins were identified. In addition, for human liver microsome analysis, 31 CYP and 16 UGT enzymes were positively identified. To achieve high throughput proteome analysis, several integrated systems were established, including multidimensional HPLC, CE and microfluidic chip based platforms. For example, with immobilized enzymatic reactor (IMER) as an effective connecting unit, 2DSEC- μRPLC for protein separation, IMER for on-line digestion, and μRPLC-ESI-MS/MS for peptides separation and identification were integrated, by which over 2000 proteins were identified from mouse liver within 20 hr. To improve the detection sensitivity of MS/MS, a series of piperazine derivatives were explored for labeling the carboxyl groups of peptides, which could be finished within a few seconds with 100% conversion ratio, and the detection sensitivity of peptides could be improved by over 10 times. All these results demonstrate that chromatography based new materials, new methods and new platforms play important roles in the high resolution, high sensitivity and high throughput analysis of proteomes.; The complexity of proteome brings great challenge and opportunity to promote the development of new techniques for liquid phase based proteome analysis. In our recent study, with newly developed methods and platforms, great improvement on proteome sample preparation, separation and identification was made. To improve the solubility of membrane proteome, and enzymatic biocompatibility with tryptic digestion, ionic liquids, BMIM BF4 showed superiority compared to the commonly applied methods, including methanol, urea and Rapist. Such a technique was successfully applied into the membrane proteome analysis of rat brain, and with the combination of 2D-HPLC-ESI-MS/MS, over 2000 non-abundant proteins, including 608 membrane proteins were identified. In addition, for human liver microsome analysis, 31 CYP and 16 UGT enzymes were positively identified. To achieve high throughput proteome analysis, several integrated systems were established, including multidimensional HPLC, CE and microfluidic chip based platforms. For example, with immobilized enzymatic reactor (IMER) as an effective connecting unit, 2DSEC- μRPLC for protein separation, IMER for on-line digestion, and μRPLC-ESI-MS/MS for peptides separation and identification were integrated, by which over 2000 proteins were identified from mouse liver within 20 hr. To improve the detection sensitivity of MS/MS, a series of piperazine derivatives were explored for labeling the carboxyl groups of peptides, which could be finished within a few seconds with 100% conversion ratio, and the detection sensitivity of peptides could be improved by over 10 times. All these results demonstrate that chromatography based new materials, new methods and new platforms play important roles in the high resolution, high sensitivity and high throughput analysis of proteomes.
文献类型会议论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/116026
专题中国科学院大连化学物理研究所
通讯作者Zhang YK(张玉奎)
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Zhang YK. New Separation and Identification Techniques for Proteome Analysis[C]. 待补充:待补充,2011:40-0.
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