Improved proteomic discovery by sample pre-fractionation using dual-column ion-exchange high performance liquid chromatography

J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Feb 15;847(1):54-61. doi: 10.1016/j.jchromb.2006.10.075. Epub 2006 Nov 30.


Clinically relevant biomarkers are urgently needed for improving patient diagnosis, risk stratification, prognosis and therapeutic treatments. There is a particularly compelling motivation for identifying protein-based indicators of early-stage disease for more effective interventions. Despite recent progress, the proteomic discovery process remains a daunting challenge due to the sheer heterogeneity and skewed protein abundances in biofluids. Even the most advanced mass spectrometry systems exhibit limiting overall dynamic ranges and sensitivities relative to the needs of modern biomedical applications. To this end, we report the development of a robust, rapid, and reproducible high performance ion-exchange liquid chromatography pre-fractionation method that allows for improved proteomic detection coverage of complex biological specimens using basic tandem mass spectrometry screening procedures. This form of sample simplification prior to global proteomic profiling, which we refer to collectively as 'fractionomics', increases the number and diversity of proteins that can be confidently identified in tissue and cell lysates as compared to the straight analysis of unfractionated crude extracts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chemical Fractionation / methods*
  • Chromatography, High Pressure Liquid / methods*
  • Humans
  • Ion Exchange
  • Mass Spectrometry / methods*
  • Mice
  • Peptide Mapping / methods*
  • Proteins / analysis*
  • Proteomics / methods*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Spectrometry, Mass, Electrospray Ionization
  • Tandem Mass Spectrometry / methods


  • Proteins