High resolution proteomics mass spectrometer with ion mobility interface coupled to nanoLC

具有与 nanoLC 耦合的离子淌度接口的高分辨率蛋白质组质谱仪

• 批准号: 499404771
• 金额: --
• 依托单位: Georg-August-Universität Göttingen
• 依托单位国家: 德国
• 项目类别: Major Research Instrumentation
• 财政年份: 2022
• 资助国家: 德国
• 起止时间: 2021-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/499404771?language=en
• 关键词: resolution; proteomics; mass; spectrometer; ion

We are applying for funding for a high resolution electrospray ionization tandem mass spectrometer equipped with an additional ion mobility-based separation stage in the interface region, and coupled to nanoflow and microflow chromatography system(s).The instrument will be installed and used in the Core Facility Proteomics of the University Medical Center Göttingen (UMG), University of Göttingen, a DFG-listed facility (RIsources RI00483) that offers mass spectrometry-based proteome analyses preferentially to all researchers of the UMG. The facility was founded in 2012 and equipped with a first generation of mass spectrometry instruments. The applied-for instrument described here will replace one of these ageing mass spectrometers installed in 2012. The applied-for instrument has to enable the following research approaches:1) data-independent acquisition mass spectrometry (DIA-MS) for medium-to-high throughput label-free analyses of cell culture, body fluid and tissue samples2) data-dependent acquisition mass spectrometry (DDA-MS) using isobaric heavy isotope labels (e.g. Tandem Mass Tags, TMT) for in-depth quantitative profiling of global protein expression and post-translational modification (PTM) states3) DDA-MS and DIA-MS analysis of chemically cross-linked protein samples for the global analysis of protein/protein interaction sites4) DDA-MS and DIA-MS analysis of protein affinity purification (AP) and protein proximity labelling experiment to identify protein/protein interactionsRequirements include an ion mobility separation stage e.g. in the interface region of the mass spectrometer to offer an additional level of selectivity for analysis of highly complex samples, and the ability to operate both nanoflow (100-350 nl min-1) and microflow (1-20 ul min-1) chromatography separations to allow for both in-depth analysis of limited sample amounts as well as high throughput analysis of larger patient/proband cohorts in the context of clinical research.

我们正在申请资助一台高分辨率电喷雾串联质谱仪，该质谱仪在界面区域配备了一个额外的基于离子迁移率的分离阶段，并与纳流和微流色谱系统耦合。该仪器将安装并使用在哥廷根大学医学中心（UMG）的核心设施蛋白质组学中，哥廷根大学，DFG列出的设施（RISTORRI 00483），优先为UMG的所有研究人员提供基于质谱的蛋白质组分析。该设施成立于2012年，配备了第一代质谱仪。本文所述的申请仪器将取代2012年安装的其中一台老化质谱仪。申请的仪器必须能够采用以下研究方法：1） 数据独立采集质谱（DIA-MS），用于细胞培养物、体液和组织样本的中高通量无标记分析2） 使用同量异位素重同位素标记（例如串联质量标签，TMT）的数据依赖性采集质谱（DDA-MS），用于全局蛋白质表达和翻译后修饰（PTM）状态的深入定量分析3） 化学交联蛋白质样品的DDA-MS和DIA-MS分析，用于蛋白质/蛋白质相互作用位点的整体分析4） 蛋白质亲和纯化（AP）的DDA-MS和DIA-MS分析和蛋白质邻近标记实验，以鉴定蛋白质/蛋白质相互作用要求包括离子迁移率分离阶段，例如在质谱仪的界面区域，以提供额外的选择性水平，用于分析高度复杂的样品，并且能够操作纳流（100-350 μ l min-1）和微流（1-20 μ l min-1）色谱分离，以允许在临床研究的背景下对有限的样品量进行深入分析以及对较大的患者/先证者群组进行高通量分析。