We have used targeted mass spectrometry–multiple reaction monitoring (MRM)–to detect the Abl biosensor and its phosphorylated form in relatively small numbers of cells. This read-out strategy enabled the scale-down of the assay, while maintaining the principle of multiplexability.
This project was funded by the NCI Innovative Molecular Analysis Technologies (IMAT) program through R21CA160129, and by the Indiana Clinical Translational Sciences Institute (ICTSI, 5TL1TR001107).
With support from the NCI IMAT program (through grant R33CA183671) and a SCIEX Young Investigator Award and Academic Partnership, we are exploring next-generation data-independent acquisition MS strategies for quantitative biosensor and biomarker profiling. Data-independent acquisition (i.e. SWATH-MS) enables high-content, quantitative analysis by using wider mass windows for analyte fragmentation–avoiding the biases that can be introduced by precursor ion selection for fragmentation.