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© Research
Publication : Methods in molecular biology (Clifton, N.J.)

Employing TMT quantification in a shotgun-MS platform

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Methods in molecular biology (Clifton, N.J.) - 01 Jan 2014

O'Brien DP, Timms JF

Link to Pubmed [PMID] – 24791989

Methods Mol. Biol. 2014;1156:187-99

The blood serum proteome may be an ideal source of disease biomarkers, although its complexity necessitates novel strategies to enrich and quantify lower-abundance proteins with biomarker utility. Herein, serum samples from pre-diagnosis pancreatic cancer cases and controls were compared using a workflow of immunodepletion, multi-lectin fractionation, and peptide tandem mass tag (TMT) labeling. Samples were then subjected to SCX and high pH reversed-phase fractionation and analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS). The aim was the discovery of candidate serum biomarkers of pancreatic cancer, although the method is applicable to any comparative proteomic analysis of serum samples.