Acquisition of OrbiTrap Eclipse w/FAIMS mass spectrometer

项目摘要/摘要： 此申请是为购买具有FAIMS质谱仪的Thermo Science Orbitrap月食提供资金 弗雷德·哈钦森癌症研究中心蛋白质组学设备的系统。该设施支持弗雷德 Hutchinson/华盛顿大学癌症联盟，由这两个机构和研究机构组成 西雅图儿童研究所和西雅图癌症护理联盟的合作伙伴，并拥有 美国国立卫生研究院的年度拨款基数超过3亿美元。我们的工厂已经运营了20年，我们有 结合75年以上的质谱学操作、维护和管理经验 仪器，以及蛋白质组学实验的设计和分析。当前基于发现的质量 该设施中的光谱仪器正在满负荷运行，导致服务等待时间很长。另外， 用于多重蛋白质组学和高灵敏度实验的设备陈旧， 但这种仪器缺乏适当支持这些实验所需的灵敏度和质量。在这 应用程序我们正在请求Orbitrap Eclipse w/FAIMS替换使用了9年的Orbitrap Elite，以便 实现两个目标：第一是增加我们工厂的样品容量/吞吐量，以减少等待时间和 第二是更换过时的仪器，以确保开展基于发现的蛋白质组学研究 在技术上适当的仪器。利用基于TMT的发现实验和分析主要 组织相容性复合体I(MHC-I)多肽是对蛋白质组学服务最频繁的请求， 基于仪器高达40赫兹的扫描速度，Orbitrap Eclipse w/FAIMS是最合适的仪器 在Orbitrap和离子陷阱中用于复杂多肽混合物的彻底采样的45赫兹，2)高离子 高灵敏度的传输和离子迁移率分离，3)实时数据库搜索，准确 多路量化，4)新的分段双曲线杆四极杆，用于更精确的TMT 量化，以及5)新的有源波束导向器，以增强灵敏度和稳健性。这是最先进的 仪器将立即支持由14笔NIH赠款资助的11名研究人员的研究，这些赠款包括 确定调节染色体分离的蛋白质和蛋白质修饰的研究，定义 与能量平衡相关的分子途径，决定了RNA突变如何拼接小体蛋白 改变细胞表型，了解与B组链球菌感染相关的毒力因素， 鉴定影响嵌合抗原受体疗效和副作用的蛋白质，并发现蛋白质 卵巢癌、乳腺癌和急性髓系白血病的生物标志物。其他项目旨在迅速确定 主要的MHC-I多肽，以更好地了解免疫系统的反应，开发针对 骨髓增生异常、白血病和其他由人乳头瘤病毒感染引起的癌症，并更好地了解 非裔美国人患者的造血干细胞移植结果。

Project Summary/Abstract: This application is for funds to purchase a Thermo Scientific Orbitrap Eclipse w/FAIMS mass spectrometry system for Fred Hutchinson Cancer Research Center’s Proteomics Facility. The facility supports the Fred Hutchinson/University of Washington Cancer Consortium which is made up of these two institutions and research partners at Seattle Children’s Research Institute and the Seattle Cancer Care Alliance and has a combined annual NIH grant base exceeding $300 million. Our facility has been operating for 20 years and we have a combined 75+ years’ experience in the operation, maintenance, and management of mass spectrometry instrumentation, and in the design and analysis of proteomics experiments. Current discovery-based mass spectrometry instrumentation in the facility is running at capacity causing long wait-times for service. Additionally, outdated instrumentation in the facility it being used for multiplexed proteomics and high-sensitivity experiments, but that instrumentation lacks the sensitivity and quality needed to properly support those experiments. In this application we are requesting the Orbitrap Eclipse w/FAIMS to replace a 9-year old Orbitrap Elite in order to achieve two goals: the first is to increase sample capacity/throughput of our facility to reduce wait-times and the second is to replace the outdated instrumentation to ensure discovery-based proteomics research is carried out on technically appropriate instrumentation. With TMT-based discovery experiments and analysis of major histocompatibility complex I (MHC-I) peptides being the most frequent requests for proteomics services, the Orbitrap Eclipse w/FAIMS is the most suitable instrument based on the instrument’s 1) scan speeds up to 40 Hz in the Orbitrap and 45 Hz in the ion trap for thorough sampling of complex peptide mixtures, 2) high ion transmission and ion mobility separations for high sensitivity, 3) real-time database searching for accurate multiplexed quantification, 4) new segmented quadrupole with hyperbolic rods for more accurate TMT quantification, and 5) novel active beam guide for enhanced sensitivity and robustness. This state-of-the art instrument would immediately support the research of 11 investigators funded by 14 NIH grants that include investigations identifying the proteins and protein modifications that regulate chromosome segregation, defining molecular pathways associated with energy balance, determining how mutations in RNA spliceosome proteins alter cellular phenotypes, understanding virulence factors associated with Group B streptococcus infection, identifying proteins that influence chimeric antigen receptor’s efficacy and side effects, and discovering protein biomarkers in ovarian cancer, breast cancer, and acute myeloid leukemia. Other projects aim to rapidly identify major MHC- I peptides to better understand immune system response, develop targeted therapies against myelodysplasia, leukemias, and other cancers driven by human papilloma virus infection, and better understand hematopoietic stem cell transplant outcomes in African American patients.