Diagnostic protein discovery using liquid chromatography/mass spectrometry for proteolytic peptide targeting.

Published

Journal Article

A peptide targeting method has been developed for diagnostic protein discovery, which combines proteolytic digestion of fractionated plasma proteins and liquid chromatography coupled to electrospray time-of-flight mass spectrometry (LC/ESI-TOFMS) profiling. Proteolysis prior to profiling overcomes molecular weight limitations and compensates for the poor sensitivity of matrix-assisted laser desorption/ionization (MALDI) protein profiling. LC/MS increases the peak capacity compared to crude fractionation techniques or single sample MALDI analysis. Differentially expressed peptides are targeted in the mass chromatograms using bioinformatic techniques and subsequently sequenced with MALDI tandem MS. In a model study comparing pancreatic cancer patients to controls, 74% of the peptide targets were successfully sequenced. This profiling method was superior to previous experiments using single sample MALDI analysis for protein profiling or proteolytic peptide profiling, because more potential protein markers were identified.

Full Text

Duke Authors

Cited Authors

  • Koomen, JM; Zhao, H; Li, D; Nasser, W; Hawke, DH; Abbruzzese, JL; Baggerly, KA; Kobayashi, R

Published Date

  • 2005

Published In

Volume / Issue

  • 19 / 12

Start / End Page

  • 1624 - 1636

PubMed ID

  • 15915451

Pubmed Central ID

  • 15915451

International Standard Serial Number (ISSN)

  • 0951-4198

Digital Object Identifier (DOI)

  • 10.1002/rcm.1963

Language

  • eng

Conference Location

  • England