Integration of Electrokinetic-Based Multidimensional Separation/Concentration Platform with Electrospray Ionization-Fourier Transform Ion Cyclotron Resonance-Mass Spectrometry for Proteome Analysis of

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• 作者：Deepa Mohan ; Ljiljana Pa//pubs.acs.org/images/entities/scaron.gif" border="0">a-Toli//pubs.acs.org/images/entities/cacute.gif" border="0"> ; Christophe D. Masselon ; Nikola
• 刊名：Analytical Chemistry
• 出版年：2003
• 出版时间：September 1, 2003
• 年：2003
• 卷：75
• 期：17
• 页码：4432 - 4440
• 全文大小：181K
• 年卷期：v.75,no.17(September 1, 2003)
• ISSN：1520-6882

文摘

This work focuses on the development of a multidimensional electrokinetic-based separation/concentration platform coupled with electrospray ionization-Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR-MS) for achieving the high resolution and ultrasensitive analysis of complex protein/peptide mixtures.A microdialysis junction is employed as the interface foron-line combination of capillary isoelectric focusing (CIEF)with transient capillary isotachophoresis/zone electrophoresis (CITP/CZE) in an integrated platform. Besidesthe excellent resolving power afforded by both CIEF andCZE separations, the electrokinetic focusing/stackingeffects of CIEF and CITP greatly enhance the dynamicrange and detection sensitivity of MS for protein identification. The constructed multidimensional separation/concentration platform is demonstrated for the analysisof Shewanella oneidensis proteome, which has considerable implications toward the bioremediation of environmental pollutants. The electrokinetic-based platform offersthe overall peak capacity comparable to those obtainedusing multidimensional chromatography systems, butwith a much shorter run time and no need for columnregeneration. Most importantly, a total of 1174 uniqueproteins, corresponding to 26.5% proteome coverage, areidentified from the cytosolic fraction of S. oneidensis,while requiring <500 ng of proteolytic digest loaded inthe CIEF capillary. The ultrasensitive capabilities ofelectrokinetic-based proteome approach are attributed tothe concentration effect in CIEF, the electrokinetic stacking of CITP, the nanoscale peak volume in CZE, the"accurate mass tag" strategy for protein/peptide identification, and the high-sensitivity, high-resolution, andhigh-mass measurement accuracy of FTICR-MS.