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

带 ETD 和 Ultimate 3000 RSLCnano 系统的 Thermo Orbitrap Eclipse Tribrid

• 批准号: 10431695
• 负责人: Shouling Xu
• 金额: $ 60万
• 依托单位: CARNEGIE INSTITUTION OF WASHINGTON, D.C.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-15 至 2023-07-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10431695
• 关键词: System

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.

该提案正在为配备 ETD 和 Ultimate 3000 RSLCnano 的 Thermo Orbitrap Eclipse Tribrid 寻求资金 系统可满足卡内基两个部门的 10 个实验室日益增长的蛋白质组分析需求 机构（九个由 NIH 资助）和斯坦福大学的三个实验室（两个由 NIH 资助）。 CISMS 配备一台仪器 Orbitrap Q Exactive HF。仅具有 HCD 模式且缺乏 MS3 功能， 由于缺乏完善的配置和软件，当前的仪器已不能满足需求 我们机构当前由 NIH 资助的研究。特别是翻译后领域许多研究人员的需求 修饰（如 O-GlcNA 酰化、O-岩藻糖基化、异戊二烯化、甲基化）、交联 质谱分析、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 目前，现有的技术无法满足映射和使用 TMT 多路复用增强量化的要求 仪器仪表。因此，计划收购 Orbitrap Eclipse ETD 对于开发和 完成了 NIH 支持的研究，因此得到了我们研究界的全力支持。拟议的 仪器将安装在卡内基科学研究所质谱设施（CIS-MS）内，该设施位于 被整个地区的研究人员广泛使用。所要求的带有 ETD 的 Orbitrap Eclipse 不仅加倍 该设施的容量，而且还提供了当前仪器所不具备的几个关键功能 该设施。特别是 ETD 和混合 EThcd 碎片模式，带有 MS3 和复杂的 配置和软件对于研究特定位点翻译后修饰的项目至关重要 分析，例如 O-GlcNAcylation 和 O-fucosylation、异戊二烯化，以及使用 MScleavable 进行结构分析 交联剂和 rRNA 转录后修饰。执行实时搜索的能力， 要求用于 TMT 同量异位定量的同步前体选择 (SPS) 和 MS3 碎片 多个小组负责蛋白质组学和高准确度、高灵敏度和高通量的项目 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 仪器将由 CIS-MS 领域经验丰富的工作人员进行。卡内基机构提供 567,739.80 美元 满足系统全价所需的匹配，并确保为设施主管和两名 支持人员以及第 2 年的服务合同以及之后仪器的剩余有效寿命 一年保修。