Improved Proteome and Phosphoproteome Analysis on a Cation Exchanger by a Combined Acid and Salt Gradient

Anal Chem. 2016 Aug 16;88(16):7899-903. doi: 10.1021/acs.analchem.6b01232. Epub 2016 Aug 2.

Abstract

Currently used elution methods for strong cation exchange (SCX) chromatography are based on two principles: salt and pH gradient. In this paper, we report the first observation of peptide elution by acid gradient. The degree of peptide separation using C18-SCX StageTip was greatly improved by our acid and salt-based elution method compared with a salt-based elution method. This development enabled us to identify over 22 000 phosphopeptides from 2 mg of protein without labor-intensive sample preparation. Our method is simple, robust, scalable, and low-cost and can be easily implemented without any special equipment or techniques.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cations / chemistry
  • Chromatography, Ion Exchange
  • HeLa Cells
  • Humans
  • Hydrogen-Ion Concentration
  • Peptides / chemistry
  • Peptides / isolation & purification*
  • Proteome*
  • Salts / chemistry

Substances

  • Cations
  • Peptides
  • Proteome
  • Salts