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学科主题: 分析化学
题名: New Separation and Identification Techniques for Proteome Analysis
作者: 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
出版日期: 2011
会议地点: 布达佩斯
通讯作者: 张玉奎
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
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.
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.
内容类型: 会议论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/116026
Appears in Collections:中国科学院大连化学物理研究所_会议论文

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Recommended Citation:
Zhang YK. New Separation and Identification Techniques for Proteome Analysis[C]. 见:36th International Symposium on High-Performance Liquid Phase Separations and Related Techniques. 布达佩斯. 2011-6-19.
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