Protein profiling by capillary isoelectric focusing, reversed-phase liquid chromatography, and mass spectrometry
In: Electrophoresis, Jg. 26 (2005-03-11), Heft 7-8
Online
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Zugriff:
An automated system for intact protein analysis is described that combines capillary isoelectric focusing (CIEF), reversed-phase liquid chromatography (RPLC), and electrospray ionization-mass spectrometry (ESI-MS). Performance is demonstrated with a complex yeast enzyme concentrate. CIEF is performed with a microdialysis membrane-based cathodic cell that permits pI fractions to be sampled and stored for subsequent LC-MS analysis. A total of 50 microg protein is loaded onto the capillary. Ten fractions are stored which span the pI range 3-10. Each fraction is subsequently cleaned on a reversed-phase trap column and then characterized by LC-MS. MaxEnt1 is used to deconvolute the raw mass spectra to obtain the molecular weight (MW) of intact proteins/peptides in the sample. A two-dimensional display of pI vs. MW is illustrated for the 500 most prevalent species as identified by MaxEnt1.
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Protein profiling by capillary isoelectric focusing, reversed-phase liquid chromatography, and mass spectrometry
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Autor/in / Beteiligte Person: | Zhou, Feng ; Johnston, Murray V. |
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Zeitschrift: | Electrophoresis, Jg. 26 (2005-03-11), Heft 7-8 |
Veröffentlichung: | 2005 |
Medientyp: | unknown |
ISSN: | 0173-0835 (print) |
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