A nano-LC system to enable functional proteomics research

***我们请求NSERC RTI计划的财政支持，以获得纳米液相色谱（nLC）单元，以与最近获得的质谱仪（MS）相结合。由此产生的nLC-MS系统将提供一个强大的技术平台，解决西部大学蛋白质组学研究在先进质谱方面的严重短缺，极大地增强了申请人团队、共同申请人及其合作者的研究计划，并使功能蛋白质组学和系统生物学的前沿研究成为可能。******用MS进行蛋白质组学分析的一个不可或缺的关键部分是在鉴定之前对分析物进行分离。最先进的LC-MS仪器包括一个高灵敏度的质量分析仪和一个液相色谱单元。生物样品的复杂性要求在质量分析之前对分析物进行物理分离。我们要求的nLC单元将能够在质谱仪鉴定之前分离微量生物样品（以纳克到毫克为单位）。在最近的CFI/ORF奖的支持下，与Li博士的癌症分子和表观遗传基础CRC （Tier I）合作，我们从ThermoFisher Scientific获得了Q-Exactive Plus (QE+) Orbitrap质谱仪。虽然QE+单元将增强我们在蛋白质组学方面的研究能力，但如果没有高分辨率的nLC单元，这种新型质谱仪的影响将无法完全实现。因此，本次RTI资金申请是购买Easy nLC1200 （ThermoFisher Scientific）来完成nLC-MS系统用于功能蛋白质组学研究。Easy nLC1200的收购不仅对QE+质谱仪的运行至关重要，而且还将显著提高我们在Western University及其他地区的蛋白质组学研究能力。使用nLC-MS平台作为在系统水平上询问蛋白质功能的工具，我们将能够揭示蛋白质表达、翻译后修饰和蛋白质-蛋白质相互作用的动态变化如何促进细胞过程（如增殖、凋亡、迁移、免疫反应、DNA损伤反应和表观遗传调节）的调节机制。此外，所要求的仪器和由此产生的nLC-MS平台将为培养功能蛋白质组学和质谱技术广泛研究领域的高素质人才（HQP）提供肥沃的土壤。* * * * * * * *

***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. ********