Selective enrichment of low-abundance peptides in complex mixtures by elution-modified displacement chromatography and their identification by electrospray ionization mass spectrometry

Anal Chem. 2002 Aug 15;74(16):3933-41. doi: 10.1021/ac025752l.

Abstract

Trace components were selectively enriched and detected in the tryptic digest of recombinant human growth hormone using elution-modified displacement chromatography, a hybrid technique combining features of elution and displacement chromatography. Based on the retention behavior of sample components in the elution mode, rapid and selective trace enrichment and high-resolution separation was achieved in a single step by utilizing appropriate combinations of an eluent such as aqueous acetonitrile with the displacer. Mass spectral and chromatographic analysis of displacement zones revealed up to 400-fold enhancement of the concentration of some low-abundance sample components. Potential application of this technique in proteomics to augment the sensitivity of LC-MS and 2-D gel electrophoretic approaches for the detection of biologically important low-abundance species is discussed.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Chromatography, High Pressure Liquid / methods*
  • Human Growth Hormone / analysis
  • Humans
  • Molecular Sequence Data
  • Peptides / analysis*
  • Recombinant Proteins / analysis
  • Sensitivity and Specificity
  • Solutions
  • Spectrometry, Mass, Electrospray Ionization
  • Trypsin / metabolism

Substances

  • Peptides
  • Recombinant Proteins
  • Solutions
  • Human Growth Hormone
  • Trypsin