Acquisition of next-generation Orbitrap Eclipse Tribrid Mass Spectrometer system

项目摘要 杜兰大学的许多NIH资助的学院需要最先进的LC-MS/MS进行生物医学研究 用于临床翻译。对这些用户的一项调查显示， 从仪器提供给杜兰大学的教职员工，而我们的研究可用的仪器发现一个 一套可以满足这些未满足需求的仪器：Orbitrap Eclipse Tribrid质量分析仪系统。的 提出的超高灵敏度Orbitrap Eclipse Tribrid LC-MS/MS系统具有几个独特的功能， 对我们的研究是必不可少的，但在我们现有的平台上，如单细胞蛋白质组学， 多级质谱（MSn）能力、增加的前体解离选项和实时搜索 同步前体选择（SPS MS 3）串联质量标签（TMT）数据采集模式。 我们已经证实，Orbitrap Eclipse Tribrid质谱仪与纳米液相色谱， UltiMate 3000 RSLCnano（U3000 LC）系统满足或超越了我们用户群的需求，并提供上级 相对于其他类似的可用LC-MS/MS系统的性能。拟议的文书将支持 发现新的生物标志物和开发具有重要生物医学意义的新方法， 我们期望在疾病诊断，机理研究和靶向治疗方面取得突破。 所提出的具有U3000 nanoLC系统的Orbitrap Eclipse Tribrid质谱法代表了下一个 LC-MS/MS系统的生成。该仪器提供更高的分辨率和更快的扫描速度， 执行并行采集以最大化占空比。我们与Thermo Scientific合作的示范项目显示， 实时搜索SPS MS 3方法将识别率提高了30%，具有更高的准确性和更少的 干扰比Q Exactive HF-x（Tulane现有质谱系统）的MS 2方法更小。的 使用Eclipse，通过数据相关采集（DDA）方法的识别率提高了76%， FAIMS Pro接口在蛋白质水平，并增加了30%，使用数据独立采集（DIA）- 基于无标记方法。所要求的电子转移解离方法可以更好地支持后处理。 翻译修改分析，更可靠的识别。新一代三杂交体MS提供了额外的 实验吞吐量，同时提供深度，覆盖范围和单细胞蛋白质组学能力。这些 性能特性将是非常有用的大型基地杜兰研究人员使用先进的LC- 用于生物医学研究的MS/MS分析。 我们已经确定了32名教员（12名主要用户和20名次要用户）资助的仪器用户群 由NIH或其他联邦赠款。他们要求利用拟议文书提供的特点， 他们的研究表明，对所要求的系统的重大需求。因此，满足这一需求是 预计将推进多个高影响力的生物医学研究项目，旨在促进公共卫生， 广泛的学科领域和NIH的研究重点。

Project Summary Many NIH-funded faculties at Tulane University require state-of-art LC-MS/MS to conduct biomedical research projects intended for clinical translation. A survey of these users revealed several features that were missing from instruments available to Tulane faculty members, while our research on available instruments found one set of instruments that could address these unmet needs: Orbitrap Eclipse Tribrid mass analyzer systems. The proposed ultra-high-sensitive Orbitrap Eclipse Tribrid LC-MS/MS system possesses several unique features that are essential for our research but are not available on our existing platform, such as single-cell proteomics, multistage mass spectrometry (MSn) capability, increased precursor dissociation options, and Real-time Search Synchronous Precursor Selection (SPS MS3) Tandem Mass Tag (TMT) data acquisition mode. We have confirmed that the Orbitrap Eclipse Tribrid mass spectrometer with nano liquid chromatography, UltiMate 3000 RSLCnano (U3000 LC) system meets or exceeds the needs of our user base and offers superior performance relative to other similar available LC-MS/MS systems. The proposed instrument will support the discovery of new biomarkers and the development of new methods of significant biomedical relevance, which we expect will lead to breakthroughs in disease diagnosis, mechanism studies, and targeted therapy. The proposed Orbitrap Eclipse Tribrid mass spectrometry with U3000 nanoLC system represents the next generation of LC-MS/MS system. The instrument provides higher resolution and faster scan speeds, and can perform parallel acquisition to maximize the duty cycle. Our demonstration project with Thermo Scientific showed that the real-time search SPS MS3 method improved identification rates by 30% with higher accuracy and less interference than the MS2 method by Q Exactive HF-x, Tulane’s existing mass spectrometry system. The identification rates via the Data-dependent acquisition (DDA) method increased by 76% using Eclipse with FAIMS Pro interface at the protein level, and increased by 30% using the Data-independent acquisition (DIA)- based label-free method. The requested electron-transfer dissociation method could better support post- translation modification analysis with more confident identification. The new generation tribrid MS gives added experimental throughput while providing depth, coverage, and single-cell proteomics capabilities. These performance characteristics will be highly useful to the large base of Tulane researchers using advanced LC- MS/MS analyses for biomedical research. We have identified an instrument user base of 32 faculty members (12 major users and 20 minor users) funded by NIH or other federal grants. They have requested the features provided by the proposed instrument to conduct their studies, demonstrating the significant need for the requested system. Meeting this need, therefore, is expected to advance multiple high-impact biomedical research projects designed to promote public health across a wide array of subject areas and NIH research priorities.