Automated 2D peptide separation on a 1D nano-LC-MS system

J Proteome Res. 2009 Mar;8(3):1610-6. doi: 10.1021/pr800986c.

Abstract

Given the complexity of the mammalian proteome, high-resolution separation technologies are required to achieve comprehensive proteome coverage and to enhance the detection of low-abundance proteins. Among several technologies, Multidimensional Protein Identification Technology (MudPIT) enables the on-line separation of highly complex peptide mixtures directly coupled with mass spectrometry-based identification. Here, we present a variation of the traditional MudPIT protocol, combining highly sensitive chromatography using a nanoflow liquid chromatography system (nano-LC) with a two-dimensional precolumn in a vented column setup. When compared to the traditional MudPIT approach, this nanoflow variation demonstrated better first-phase separation leading to more proteins being characterized while using rather simple instrumentation and a protocol that requires less time and very little technical expertise to perform.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cells, Cultured
  • Chromatography, Liquid / instrumentation
  • Chromatography, Liquid / methods*
  • Mice
  • Myocytes, Cardiac / metabolism*
  • Nanotechnology
  • Proteome / metabolism*
  • Tandem Mass Spectrometry / methods*

Substances

  • Proteome