MULTI-REPORTER ISOBARIC LABELING REAGENTS FOR INCREASED MULTIPLEXING IN QUANTITATIVE PROTEOMICS

Project SummaryThe objective of this proposal is to develop next generation isobaric labeling reagents that will greatly expand the utility of multiplexed quantitative proteomics and its applications in clinical and pre clinical biomedical researchThe isobaric labeling strategy involves a set of chemical reagents for tagging peptides and other biomolecules with a stable isotope encoded barcodeThe ability to mix and analyze multiple barcoded samples simultaneously without increasing LC MS complexityThis preserves analytical depthreduces experimental variabilityand increases throughputThis technology is extending the boundaries of proteomics into more profound areas of primary and clinical researchwith incredibly powerful and far ranging applications reported in areas such as drug target identificationbiomarker discoveryand temporal regulation of proteome dynamicsWhile isobaric labeling has been established as an accuratereliableand sensitive quantitative techniquethere is a definitive need for improvement in isobaric multiplexing capacityThefold multiplexing of current isobaric labeling reagents limits analysis to relatively simple experimental designsand is insufficient for the large cohort analyses required to achieve biological insight in the context diversity among the human populationThe reagents proposed herein employ a novel mechanism that involves spontaneous formation of multiple barcodes per reagentproviding the ability to encode additional informationvia heavy isotopeswhich ultimately expands the multiplexing capacity these reagents for quantitative proteomicsThis proposal will address key remaining hurdles to commercialization of these reagents byincreasing reagent performance through the development of optimized LC MS methodsrefining data analysis methods to improve measurement precisiondeveloping turnkey software for seamless integration of the approach into current proteomics workflowsusing computer modeling to establish an optimal prototypePlex reagent setsynthesis and validation of this prototype reagent set across a variety of applicationsandestablish a synthesis strategy that is scalable to commercial demand Narrative Quantitative proteomics is an emerging tool for both pre clinical and clinical research and developmentbut limitations in throughput and sensitivity hampers widespread utilityThis work will develop a next generation technology for multiplexed quantitative proteomics withfold improvement in multiplexing capacity over current methodsOur unique innovation greatly improves the throughput of quantitative proteomicsproviding new avenues for accuratesensitive and reproducible interrogation of clinically relevant biology!