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Fully automatable two-dimensional reversed-phase capillary liquid chromatography with online tandem mass spectrometry for shotgun proteomics
Siu S.O.1; Lam M.P.Y.1; Lau E.1; Kong R.P.W.1; Lee S.M.Y.2; Chu I.K.1
2011-06-01
Source PublicationProteomics
ISSN16159853 16159861
Volume11Issue:11Pages:2308-2319
Abstract

Herein, we describe the development of a fully automatable technology that features online coupling of high-pH RP separation with conventional low-pH RP separation in a two-dimensional capillary liquid chromatography (2-D LC) system for shotgun proteomics analyses. The complete analysis comprises 13 separation cycles, each involving transfer of the eluate from the first-dimension, high-pH RP separation onto the second RP dimension for further separation. The solvent strength increases across the 13 fractions (cycles) to elute all peptides for further resolution on the second-dimension, low-pH RP separation, each under identical gradient-elution conditions. The total run time per analysis is 52h. In triplicate analyses of a lysate of mouse embryonic fibroblasts, we used this technology to identify 2431 non-redundant proteins, of which 50% were observed in all three replicates. A comparison of RP-RP 2-D LC and strong cation exchange-RP 2-D LC analyses reveals that the two technologies identify primarily different peptides, thereby underscoring the differences in their separation chemistries. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

KeywordLc-ms/ms Multidimensional Lc Protein Profile Technology
DOI10.1002/pmic.201100110
URLView the original
Indexed BySCI
WOS Research AreaBiochemistry & Molecular Biology
WOS SubjectBiochemical Research Methods ; Biochemistry & Molecular Biology
WOS IDWOS:000291087700014
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被引频次[WOS]:20   [WOS记录]     [WOS相关记录]
Document TypeJournal article
专题Institute of Chinese Medical Sciences
Affiliation1.The University of Hong Kong
2.University of Macau
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Siu S.O.,Lam M.P.Y.,Lau E.,et al. Fully automatable two-dimensional reversed-phase capillary liquid chromatography with online tandem mass spectrometry for shotgun proteomics[J]. Proteomics,2011,11(11):2308-2319.
APA Siu S.O.,Lam M.P.Y.,Lau E.,Kong R.P.W.,Lee S.M.Y.,&Chu I.K..(2011).Fully automatable two-dimensional reversed-phase capillary liquid chromatography with online tandem mass spectrometry for shotgun proteomics.Proteomics,11(11),2308-2319.
MLA Siu S.O.,et al."Fully automatable two-dimensional reversed-phase capillary liquid chromatography with online tandem mass spectrometry for shotgun proteomics".Proteomics 11.11(2011):2308-2319.
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