title: Optimal selection of 2D reversed-phase-reversed-phase HPLC separation techniques in bottom-up proteomics.
abstract: Recent developments in bottom-up proteomics have supplanted the use of gel-based approaches in favor of multidimensional chromatographic separations of peptide mixtures followed by mass spectrometry analysis. This trend is driven by the desire to eliminate labor-intensive in-gel digestion procedures and increase proteome coverage through better recovery of proteolytic fragments. Introduction of reversed-phase-reversed-phase 2D separation techniques is one of the major improvements that have made this possible. In this article, we review recent developments in 2D HPLC and highlight variations in reversed-phase HPLC separation selectivity that allow for superior peak capacity in peptide fractionation.
authors.affiliation: Manitoba Centre for Proteomics & Systems Biology, University of Manitoba, 715 McDermot Avenue, Winnipeg, MB, Canada.
authors.forename: Peter
authors.initials: P
authors.lastname: McQueen
authors.forename: Oleg
authors.initials: O
authors.lastname: Krokhin