A double-vented tetraphasic continuous column approach to MuDPIT analysis on long capillary columns demonstrates superior proteomic coverage

J Proteome Res. 2005 Nov-Dec;4(6):2412-9. doi: 10.1021/pr050209h.

Abstract

A double-vented serial tetraphasic capillary column approach is applied to proteomic MuDPIT-type analysis using extended length capillary reverse-phase columns. The heart of the tetraphasic device consists of a triphasic MuDPIT trap located upstream of a venting tee. The trap is followed by a 60 cm high-resolution capillary column. A conventional high-flow HPLC is used to develop gradients at standard flow rates and pressures. The double-vented triphasic MuDPIT trapping device relieves the capillary separation column from the salt burden during the on-line cation-exchange portion of the analysis. Two configurations are presented, a double-vented continuous column model and a discontinuous model in which the triphasic MuDPIT trap is installed on a six-port valve; both configurations were tested with 60 and 10 cm capillary columns. All four systems were challenged with a trypsin digest of undepleted human serum, and a matrix of proteomic results for the different models and column lengths are compared.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blood Proteins / chemistry
  • Cations
  • Chromatography
  • Chromatography, High Pressure Liquid
  • Chromatography, Ion Exchange
  • Electrophoresis, Capillary
  • Humans
  • Mass Spectrometry
  • Nanotechnology
  • Peptides / chemistry
  • Proteome
  • Proteomics / instrumentation*
  • Proteomics / methods*
  • Time Factors
  • Trypsin / pharmacology

Substances

  • Blood Proteins
  • Cations
  • Peptides
  • Proteome
  • Trypsin