A peptide preconcentration approach for nano-high-performance liquid chromatography to diminish memory effects

An advanced method has been developed for the analysis of proteolytic digests of complex protein mixtures by reversed phase high‐performance liquid chromatography coupled to mass spectrometry. The occurrence of memory effects was prevented by a parallel set of two precolumns employed for simultaneou...

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Bibliographic Details
Published in:Proteomics (Weinheim) 2004-09, Vol.4 (9), p.2541-2544
Main Authors: Schaefer, Heike, Chervet, Jean-Pierre, Bunse, Christian, Joppich, Cornelia, Meyer, Helmut E., Marcus, Katrin
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Chromatography, High Pressure Liquid - instrumentation
Chromatography, High Pressure Liquid - methods
Complex peptide mixtures
Fundamental and applied biological sciences. Psychology
Mass Spectrometry - methods
Memory effect
Miscellaneous
Nano-separations
Nanotechnology - instrumentation
Nanotechnology - methods
Peptide preconcentration
Peptides - analysis
Proteins
Reserved-phase high-performance liquid chromatography
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Summary:An advanced method has been developed for the analysis of proteolytic digests of complex protein mixtures by reversed phase high‐performance liquid chromatography coupled to mass spectrometry. The occurrence of memory effects was prevented by a parallel set of two precolumns employed for simultaneous separation and washing procedures. The system was tested extensively, and tryptic digests of three single proteins were analyzed. In addition, different solvent systems were evaluated for effective washing of the employed precolumns. Using the analytical strategy presented, a reliable identification of proteins in complex mixtures was obtained and not hampered by the occurrence of memory effects.
ISSN:1615-9853
1615-9861
DOI:10.1002/pmic.200300801