Reference : Highly Multiplexed Targeted Proteomics Acquisition on a TIMS-QTOF
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
Systems Biomedicine
Highly Multiplexed Targeted Proteomics Acquisition on a TIMS-QTOF
Lesur, Antoine [Luxembourg Institute of Health - LIH > Quantitative Biology Unit]
Schmit, Pierre-Olivier [Bruker Daltonics S.A., France]
Bernardin, François [Luxembourg Institute of Health - LIH > Quantitative Biology Unit]
Letellier, Elisabeth mailto [University of Luxembourg > Faculty of Science, Technology and Medicine (FSTM) > Department of Life Sciences and Medicine (DLSM) >]
Brehmer, Sven [Bruker Daltonik GmbH, Bremen, Germany]
Decker, Jens [Bruker Daltonik GmbH, Bremen, Germany]
Dittmar, Gunnar [Luxembourg Institute of Health - LIH > Quantitative Biology Unit]
Analytical Chemistry
American Chemical Society
Yes (verified by ORBilu)
[en] Targeted proteomics allows the highly sensitive detection of specific peptides and proteins in complex biological samples. Here, we describe a methodology for targeted peptide quantification using a trapped ion mobility quadrupole time-of-flight mass spectrometer (timsTOF Pro). The prm-PASEF method exploits the multiplexing capability provided by the trapped ion mobility separation, allowing more than 200 peptides to be monitored over a 30 min liquid chromatography separation. Compared to conventional parallel reaction monitoring (PRM), precursor ions are accumulated in the trapped ion mobility spectrometry (TIMS) cells and separated according to their shape and charge before eluting into the quadrupole time-of-flight (QTOF) part of the mass spectrometer. The ion mobility trap allows measuring up to six peptides from a single 100 ms ion mobility separation with the current setup. Using these improved mass spectrometric capabilities, we detected and quantified 216 isotope-labeled synthetic peptides (AQUA peptides) spiked in HeLa human cell extract with limits of quantification of 17.2 amol for some peptides. The acquisition method is highly reproducible between injections and enables accurate quantification in biological samples, as demonstrated by quantifying KRas, NRas, and HRas as well as several Ras mutations in lung and colon cancer cell lines on fast 10 min gradient separations.
FnR ; FNR11282028 > Elisabeth Letellier > miRMet > Role Of Mir-371-373 Cluster In Tumor Initiation And Metastatic Colonization > 15/03/2017 > 14/07/2020 > 2016

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