Acquisition of next-generation Orbitrap Eclipse Tribrid Mass Spectrometer system

项目摘要 杜兰大学许多由美国国立卫生研究院资助的学院需要最先进的LC-MS/MS来进行生物医学研究 面向临床翻译的项目。对这些用户的一项调查显示，有几项功能缺失 来自杜兰大学教职员工可用的仪器，而我们对可用的仪器的研究发现了一个 一套可以解决这些未得到满足的需求的仪器：Orbitrap、Eclipse、Tribrid质谱仪系统。这个 提出的超高灵敏Orbitrap月食三元LC-MS/MS系统具有几个独特的特征 对于我们的研究是必不可少的，但在我们现有的平台上是不可用的，例如单细胞蛋白质组学， 多级质谱学(MSN)功能、增加的前体解离选项和实时搜索 同步前体选择(SPS MS3)串联质量标签(TMT)数据采集模式。 我们已经证实了Orbitrap Eclipse三元联用纳米液相色谱质谱仪， 旗舰3000 RSLCNano(U3000 LC)系统满足或超过我们用户群的需求，并提供卓越的 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 新生物标志物的发现和具有重大生物医学意义的新方法的开发，这 我们预计将在疾病诊断、机制研究和靶向治疗方面取得突破。 建议使用U3000 NanoLC系统的Orbitrap Eclipse三联质谱仪代表了下一步 LC-MS/MS系统的产生。该仪器提供更高的分辨率和更快的扫描速度，并且可以 执行并行采集以最大限度地提高占空比。我们与Thermo Science的示范项目显示 实时搜索SPS MS3方法识别率提高30%，准确率更高，识别率更低 比QExactive HF-x(杜兰现有的质谱学系统)使用的MS2方法更容易受到干扰。这个 通过数据依赖获取(DDA)方法实现的识别率在使用 FAIMS Pro接口，并使用数据独立采集(DIA)增加了30%- 基于无标签的方法。所要求的电子转移解离方法可以更好地支持后 翻译修改分析，更有把握地识别。新一代Tribrid MS增加了 实验吞吐量，同时提供深度、覆盖率和单细胞蛋白质组学能力。这些 性能特征将对使用先进LC的杜兰大学的大量研究人员非常有用。 用于生物医学研究的MS/MS分析。 我们已经确定了一个由32名教职员工(12名主要用户和20名次要用户)资助的仪器用户基础 由美国国立卫生研究院或其他联邦拨款。他们已要求实施拟议文书所提供的功能 他们的研究，表明了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，旨在促进全社会的公共健康 广泛的学科领域和美国国立卫生研究院的研究重点。

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.