The new tims-TOF pro-mass spectrometer offers improved capabilities for the targeted acquisition. The prm-PASEF method uses 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 TIMS cells and separated according to their shape and charge before eluting into the Q-TOF part of the mass spectrometer. 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 method's applicability is demonstrated by measuring Kras, Nras, and Hras in biological samples and the most common cancer-associated Kras mutations.
Integrated TIMS with PASEF affords an opportunity for highly selective, rapid detection and reproducible quantification of peptides in complex matrices. Here we describe PASEF-PRM-LIVE, a framework implementing on-the-fly adjustment of the detection window to maximize target coverage. This platform is ideally suited for functional protein profiling.
Prof. Dr. Gunnar Dittmar
Group leader proteomics of cellular signalling
Luxembourg Institute of Health, Department of Infection and Immunity
Jarrod A. Marto, Ph.D.
Associate Professor, Dept. of Pathology
Brigham and Women’s Hospital and Harvard Medical School, Dana-Farber Cancer Institute
For Research Use Only. Not for use in clinical diagnostic procedures.