Journal
RAPID COMMUNICATIONS IN MASS SPECTROMETRY
Volume 20, Issue 12, Pages 1932-1938Publisher
WILEY
DOI: 10.1002/rcm.2541
Keywords
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A system of microchip capillary electrophoresis/electrospray ionization mass spectrometry (mu chip-CE/ESI-MS) for rapid characterization of proteins has been developed. Capillary electrophoresis (CE) enables rapid analysis of a sample present in very small quantity, such as at femtomole levels, at high resolution. Faster CE/MS analysis is expected by downsizing the normal capillary to the microchip (mu chip) capillary. Although rapidity and high resolution are advantages of CE separation, electro-osmotic flow (EOF) instability caused by the interaction between proteins and the microchannel surface results in low reproducibility in the analysis of basic proteins under neutral pH conditions. By coating the microchannel surface with a basic polymer, polyE-323, basic proteins, which have pI values of over 7.5, could be separated and detected by mu chip-CE/MS on quadrupole (Q) and time-of-flight (TOF) hybrid instruments. By increasing the cone and collision voltages during the analysis by mu chip-CE/ESI-MS of a small protein, some product ions, which contain the sequence information, could also be obtained, i.e., 'top-down' analysis of the protein could be accomplished with this mu chip-CE/MS system. To our knowledge, this is the first report of 'top-down' analysis of a protein by mu chip-CE/MS. Since it requires a much shorter time and a smaller sample amount for analysis than the conventional liquid chromatography (LC)/ESI-MS method, mu chip-CE/MS promises to be suitable for the high-throughput characterization of proteins. Copyright (c) 2006 John Wiley & Sons, Ltd.
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