on 07-10-201310:10 AM - edited on 10-15-202111:38 AM by Closed Account
Blank M, Bomgarden R, Rogers J, Jacobs R, Fong J, Puri N, Zabrouskov V, Viner R. ASMS 2013 Poster Note Purpose: To determine the mechanism of tyrosine kinase inhibitor (TKI) resistance through analysis of differential changes in protein pathway expression levels by liquid chromatography-mass spectrometry (LC-MS).
Methods: Parental and tyrosine-kinase inhibitor (Erlotinib) resistant cell lines were treated with Epidermal Growth Factor (EGF), Erlotinib, Erlotinib and EGF, or left untreated. Samples from each condition were labeled with TMT8plex isobaric tags and analyzed/quantified by LC-MS on two hybrid instruments based on the Thermo Scientific™ Orbitrap™ mass analyzer.
Results: The increased multiplexing capabilities of Tandem Mass Tag® (TMT®) labeling enables simultaneous comparison of 8 different treatment conditions. Changes in protein expression ratios between parent and TKI-resistant cell lines were observed in several important cell signaling pathways with improved quantitative accuracy using synchronous precursor selection (SPS) acquisition methods compared to higher-energy collision dissociation (HCD) MS2 methods.