Thermo Orbitrap Eclipse Tribrid with ETD and an Ultimate 3000 RSLCnano System

该提案正在寻求资金，用于配备ETD的Thermo Orbitrap月食三合一和旗舰3000 RSLCNano 满足卡内基两个系10个实验室日益增长的蛋白质组分析需求的系统 斯坦福大学的三个实验室(两个由美国国立卫生研究院资助)。信息与通信技术中心 配备Orbitrap Q Exactive HF一台仪器。由于只有HCD模式并且缺乏MS3功能， 缺乏复杂的配置和软件，目前的仪器已不能满足需求 我们机构目前由NIH资助的研究。特别是，许多研究人员在翻译后领域的需求 修饰(如O-GlcN酰化、O-岩藻糖化、戊烯基化、甲基化)、交联化 质谱学，细胞类型特异性和单细胞蛋白质组学，rRNA修饰，蛋白质-RNA界面 映射和使用TMT多路复用的增强型量化，目前没有满足现有的 仪器仪表。因此，收购Orbitrap Eclipse ETD的计划对开发和 完成了NIH支持的研究，因此得到了我们研究界的全力支持。建议数 仪器将安装在卡内基科学研究所的质谱学设施(CIS-MS)内，这是 被整个地区的研究人员高度使用。请求的带有ETD的Orbitrap Eclipse不仅将 设施的能力，但也提供了几个关键的能力，这是目前的工具 这个设施。特别是，ETD和混合EThcd碎片模式、MS3和高级 配置和软件，对于研究特定站点翻译后修改的项目来说是必不可少的 对O-GlcN酰化、O-岩藻糖化、戊烯基化等进行分析，并利用MScleavable进行结构分析 交联剂和rRNA转录后修饰。能够执行实时搜索， 用于TMT等压定量的同步前体选择(SPS)和MS3碎片的请求来自 要求蛋白质组学高精度、高灵敏度和高通量的项目的几个小组 磷酸蛋白质组学、细胞类型蛋白质组学和单细胞蛋白质组学。操作和维护 乐器将由独联体-MS的经验丰富的工作人员演奏。卡内基研究所提供了567,739.80美元 需要匹配以满足系统的全价，并确保为设施主管和两名 支持人员，以及第二年的服务合同和仪器的剩余有效寿命 一年的保修期。

This proposal is seeking funds for a Thermo Orbitrap Eclipse Tribrid with ETD and an Ultimate 3000 RSLCnano system to meet increasing demands for proteomic analysis for 10 labs from two departments of Carnegie Institution (nine are funded by NIH) and for three labs at Stanford University ( two are funded by NIH). The CISMS is equipped with one instrument Orbitrap Q Exactive HF. With only HCD mode and lack of MS3 functionality, lack of sophisticated configuration and software, the current instrument can no longer meet the demands of current NIH-funded research at our institution. In particular, the needs of many researchers in the area of posttranslational modifications (such as O-GlcNAcylation, O-fucosylation, prenylation, methylation), cross-linking mass spectrometry, cell-type specific and single-cell proteomics, rRNA modification, protein-RNA interface mapping, and enhanced quantification using TMT multiplexing, are currently not being met with existing instrumentation. Thus, the planned acquisition of Orbitrap Eclipse ETD is crucial for the development and completion of NIH-supported research, and thereby is fully backed by our research community. The proposed instrument is to be housed within Carnegie Institution for Science Mass Spectrometry facility(CIS-MS) which is highly used by researchers throughout the region. The requested Orbitrap Eclipse with ETD not only doubles the capacity of the facility, but also provides several key capabilities not available on the current instrument of the facility. In particular, ETD and hybrid EThcd fragmentation modes, with MS3 and with sophisticated configuration and software, are essential for projects that study site-specific post-translational modification analysis such as O-GlcNAcylation and O-fucosylation, and prenylation, and structural analysis using MScleavable cross-linker, and rRNA post-transcriptional modifications. The ability to perform real-time search, synchronous precursor selection (SPS) and MS3 fragmentation for TMT isobaric quantification is requested from several groups for projects that demand high accuracy, high sensitivity and high throughput for proteomics and phosphoproteomics, cell type proteomics and single-cell proteomics. Operation and maintenance of the instrument will be performed by experienced staff in CIS-MS. The Carnegie institution provides the $567,739.80 match required to meet the full price of the system, and ensures full support for the facility director and two supporting staff, as well as service contract for year 2 and the remaining effective lifetime of the instrument after the one-year warranty.