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Combining capillary electrophoresis with mass spectrometry for applications in proteomics
Author(s) -
Simpson David C.,
Smith Richard D.
Publication year - 2005
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200410132
Subject(s) - chromatography , chemistry , capillary electrophoresis , mass spectrometry , capillary electrophoresis–mass spectrometry , isoelectric focusing , proteomics , bottom up proteomics , analyte , protein mass spectrometry , tandem mass spectrometry , electrospray ionization , biochemistry , gene , enzyme
Abstract Mass spectrometry (MS)‐based proteomics is currently dominated by the analysis of peptides originating either from digestion of proteins separated by two‐dimensional gel electrophoresis (2‐DE) or from global digestion; the simple peptide mixtures obtained from digestion of gel‐separated proteins do not usually require further separation, while the complex peptide mixtures obtained by global digestion are most frequently separated by chromatographic techniques. Capillary electrophoresis (CE) provides alternatives to 2‐DE for protein separation and alternatives to chromatography for peptide separation. This review attempts to elucidate how the most promising CE modes, capillary zone electrophoresis (CZE) and capillary isoelectric focusing (CIEF), might best be applied to MS‐based proteomics. CE‐MS interfacing, mass analyzer performance, column coating to minimize analyte adsorption, and sample stacking for CZE are considered prior to examining numerous applications. Finally, multidimensional systems that incorporate CE techniques are examined; CZE often finds use as a fast, final dimension before ionization for MS, while CIEF, being an equilibrium technique, is well‐suited to being the first dimension in automated fractionation systems.