|Sarah Michaud received her M.Sc. in Genome Sciences and Technology at the University of British Columbia, where she investigated quantitative proteomics of honeybees during viral infection. She began her work developing targeted assays for quantitation of mouse plasma proteins at MRM Proteomics in 2014, and is now a project manager at the UVic Genome BC Proteomics Center in Victoria, Canada, where her focus is the development of 3,000 MRM assays in 20+ mouse tissues. |
Development of Quantitative MRM Assays for the Measurement of 3,000 Proteins across 20 Mouse TissuesAn ongoing effort at our Centre is the development of MRM-MS assays quantifying 3,000 proteins across 20 mouse tissues for use in molecular phenotyping. Our assays use stable isotope-labelled standard peptides for quantitation and undergo rigorous optimization of peptide- and instrument-specific parameters followed by assay development and validation according to CPTAC guidelines. Assay development determines the LLOQ and linear range of the assay via standard curve, and assay variability is calculated over 5 days at three concentrations spanning the final range of the assays. Read More
To date assays have been developed and validated for 1643 unique proteins in 11 mouse tissues. Once developed, assays are multiplexed into panels of 125 proteins and used to measure normal tissue protein concentrations in 3 common mouse strains. Data processing and analysis for all steps, from optimization to sample measurement, was performed using Skyline, and is a critical step in our pipeline for large scale assay development.