Automated on-line ionic detergent removal from minute protein/peptide samples prior to liquid chromatography-electrospray mass spectrometry

Journal of Chromatography. B, Biomedical Applications
J P VissersC A Cramers

Abstract

An automated on-line ionic detergent removal pre-column system coupled to capillary liquid chromatography-electrospray mass spectrometry is described. The system involves two micro precolumns, composed of a specific ionic detergent trapping column and a preconcentration column, respectively, and a packed 300 microns I.D. analytical column. Sample loading to the micro precolumns and regeneration of the detergent trapping column were performed at a flow-rate of 50 microliters/min, while the flow-rate through the analytical column was set at 5.0 microliters/min. Ionic detergent-containing tryptic-digested protein samples were directly applied to the micro precolumns without sample pretreatment and were analysed by UV absorption detection and electrospray mass spectrometry. The presented system allows for the fully automated removal of SDS with virtually no loss in protein/peptides. Maximum SDS load and breakthrough have been determined. Excellent protein recovery and complete removal of SDS is found. The chromatographic separation after SDS removal was completely restored and equalled the reference chromatogram. Mass spectral data confirm these findings. Finally, this technique allows for SDS removal from minute protein samples wi...Continue Reading

References

Jul 1, 1988·Analytical Biochemistry·H Suzuki, T Terada
Dec 25, 1987·Journal of Chromatography·B Kaplan, M Pras
Sep 15, 1981·Analytical Biochemistry·R Amons, P I Schrier

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Citations

Oct 6, 2000·Analytical Biochemistry·E Yang-BojaH M Fales
Aug 1, 2002·Journal of the American Society for Mass Spectrometry·Johannes P C VissersM Arthur Moseley
Jan 7, 2011·PloS One·Chandra Sekhar Rao KadiyalaMasaru Miyagi
Oct 12, 2010·Mass Spectrometry Reviews·Maroun Bou KhalilDaniel Figeys
Jul 12, 2007·Journal of Separation Science·An StaesKris Gevaert
Nov 18, 2011·Expert Review of Proteomics·Alan A DoucetteShayla Fitzsimmons

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