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 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.
Schaefer
Heike
H
Medical Proteom-Center, Ruhr-University of Bochum, Bochum, Germany.
Chervet
Jean-Pierre
JP
Bunse
Christian
C
Joppich
Cornelia
C
Meyer
Helmut E
HE
Marcus
Katrin
K
D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D036103:Nanotechnology / Q000295:instrumentation / Q000379:methods; D010455:Peptides / Q000032:analysis*
D010455:Peptides