A nano-LC system to enable functional proteomics research

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***We request financial support from the NSERC RTI program to obtain a nano-Liquid Chromatography (nLC) unit to be coupled with a recently acquired mass spectrometer (MS). The resulting nLC-MS system will provide a robust technological platform that will address a critical shortage in advanced mass spectrometry for proteomic research at Western University, drastically enhance the research programs for the team of applicant, co-applicants and their collaborators, and enable cutting-edge research in functional proteomics and systems biology. ******An integral and critical part of proteomics analysis by MS is the separation of the analytes before their identification. The state-of-the-art instrumentation for LC-MS comprises a highly sensitive mass analyzer coupled at the front with a liquid chromatography unit. The complexity of biological samples demands that the analytes be physically separated prior to mass analysis. The nLC unit that we are requesting will enable separation of a minute amount of biological sample (in nanograms-to-milligrams) prior to its identification in the mass spectrometer. Under the auspices of a recent CFI/ORF award in association with Dr. Li's CRC (Tier I) in Molecular and Epigenetic Basis of Cancer, we acquired a Q-Exactive Plus (QE+) Orbitrap mass spectrometer from ThermoFisher Scientific. While the QE+ unit will enhance our research capacity in proteomics, the impact of this new mass spectrometer will not be fully realized without a high-resolution nLC unit. Therefore, this RTI funding request is to purchase an Easy nLC1200 (ThermoFisher Scientific) to complete the nLC-MS system for functional proteomics research. The acquisition of the Easy nLC1200 is not only essential for the operation of the QE+ mass spectrometer, it will also markedly increase our research capacity in proteomics at Western University and beyond. Using the nLC-MS platform as a tool to interrogate protein function at the systems level, we will be able to uncover the mechanisms behind how dynamic alterations in protein expression, post-translational modification and protein-protein interaction contribute to the regulation of cellular processes such as proliferation, apoptosis, migration, immune response, DNA damage response, and epigenetic regulation. Furthermore, the requested instrumentation and the resulting nLC-MS platform will provide a fertile ground for the training of highly qualified personnel (HQP) in functional proteomics and a wide range of research areas enabled by mass spectrometry. ********