DICP OpenIR
Subject Area物理化学
Multi-dimensional capillary liquid chromatography-tandem mass spectrometry for phosphoproteome analysis
Zou HF(邹汉法); Wang FJ(王方军); Ye ML(叶明亮); Han GH(韩广辉); Jiang XN(江新宁)
Conference NameThe 23th International Symposium on Micro-scale Bioseparation
Conference Date2009-2-1
2009-02-01
Conference Place美国
Pages251/1
Department十八室
Funding OrganizationThe California Separation Science Society
AbstractPhosphorylated peptides enriched from cancerous human liver are very complex and the amount is always limited. It is important to develop powerful fractionating method to increase the identification of low abundance phosphorylated peptides by decreasing sample complexity. A new approach was established by using off-line multi-dimensional capillary liquid chromatography-tandem mass spectrometry to increase the identification coverage of human liver phosphoproteome analysis. The tryptic digest of human liver extract firstly was pretreated with Ti –IMAC adsorbent, and then the enriched mixture was further fractionated by using a 9 cm × 200 µm i.d. strong anion-exchange (SAX) capillary trap column. The pass-through fraction was loaded onto C18 trap column for capillary liquid chromatography-tandem mass spectrometry analysis, and then the SAX trap column with adsorbed components was further coupled to C18 analytical column for multi-dimensional capillary liquid chromatography-tandem mass spectrometry analysis. The good complementarity for identification of phosphopeptides among the pass-through fraction and adsorbed components with SAX capillary trap column was observed and the total number of the identified phosphopeptides increased 105.6% by comparing to conventional method by directly loading Ti-IMAC enriched fraction onto C18 trap column for capillary liquid chromatography-tandem mass spectrometry analysis. When 500 µg tryptic digest of human liver extract was analyzed by this new approach with 300-min reversed phase gradient separation, totally 876 unique phosphorylated peptides (among which 224 were multiple phosphorylated peptides) and 926 phosphorylated sites were positively identified from 494 phosphorylated proteins at FDR 0.92%.
Language中文
Document Type会议论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/114006
Collection中国科学院大连化学物理研究所
Corresponding AuthorZou HF(邹汉法)
Recommended Citation
GB/T 7714
Zou HF,Wang FJ,Ye ML,et al. Multi-dimensional capillary liquid chromatography-tandem mass spectrometry for phosphoproteome analysis[C],2009:251/1.
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