biostudies-literatureBritton DCancer Research UKMedical Research CouncilNational Institute for Health Research (NIHR)e90948https://www.ebi.ac.uk/biostudies/studies/S-EPMC3966770biostudies-literatureUnknown9(3)<h4>Objective</h4>LC-MS/MS phospho-proteomics is an essential technology to help unravel the complex molecular events that lead to and propagate cancer. We have developed a global phospho-proteomic workflow to determine activity of signaling pathways and drug targets in pancreatic cancer tissue for clinical application.<h4>Methods</h4>Peptides resulting from tryptic digestion of proteins extracted from frozen tissue of pancreatic ductal adenocarcinoma and background pancreas (n = 12), were labelled with tandem mass tags (TMT 8-plex), separated by strong cation exchange chromatography, then were analysed by LC-MS/MS directly or first enriched for phosphopeptides using IMAC and TiO2, prior to analysis. In-house, commercial and freeware bioinformatic platforms were used to identify relevant bPloS oneQuantification of pancreatic cancer proteome and phosphorylome: indicates molecular events likely contributing to cancer and activity of drug targets.PMC3966770NIHR-RP-011-0531605714549MR/J006742/1Nicholson RSarker DPike IGee JQuaglia AWard MSelzer SStebbing JZucht HDHoehle CSchmid PKoncarevic SBohm GPrefot PMitra VCastellano LHeaton NLoβner CBritton DJung SZen YfalseQuantification of pancreatic cancer proteome and phosphorylome: indicates molecular events likely contributing to cancer and activity of drug targets.<h4>Objective</h4>LC-MS/MS phospho-proteomics is an essential technology to help unravel the complex molecular events that lead to and propagate cancer. We have developed a global phospho-proteomic workflow to determine activity of signaling pathways and drug targets in pancreatic cancer tissue for clinical application.<h4>Methods</h4>Peptides resulting from tryptic digestion of proteins extracted from frozen tissue of pancreatic ductal adenocarcinoma and background pancreas (n = 12), were labelled with tandem mass tags (TMT 8-plex), separated by strong cation exchange chromatography, then were analysed by LC-MS/MS directly or first enriched for phosphopeptides using IMAC and TiO2, prior to analysis. In-house, commercial and freeware bioinformatic platforms were used to identify relevant b2014-01-01T00:00:00Z20142026-04-29T21:11:54.144Z2019-03-26T23:24:23ZS-EPMC39667702467041610.1371/journal.pone.0090948