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

我们正在申请资助一台高分辨率电喷雾电离串联质谱仪，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(S)耦合。该仪器将安装并用于哥廷根大学医学中心(UMG)的核心设施蛋白质组学，这是一家在DFG上市的设施(RIsources RI00483)，优先向UMG的所有研究人员提供基于质谱学的蛋白质组分析。该设施成立于2012年，配备了第一代质谱仪。这里介绍的申请仪器将取代2012年安装的其中一个老化的质谱计。所应用的仪器必须实现以下研究方法：1)用于细胞培养、体液和组织样本的中高通量无标记分析的数据独立采集质谱学(DIA-MS)；2)使用等压重同位素标记(例如串联质量标签，TMT)用于深入定量描述全球蛋白质表达和翻译后修饰(PTM)状态3)用于蛋白质/蛋白质相互作用部位的全球分析的化学交联蛋白质样品的DDA-MS和DIA-MS分析4)蛋白质亲和纯化(AP)的DDA-MS和DIA-MS分析以及用于识别蛋白质/蛋白质相互作用的蛋白质邻近标记实验要求包括例如在质谱仪的界面区域中的离子迁移率分离阶段，以为高度复杂的样品的分析提供额外的选择性水平，能够同时进行纳流(100-350 nL min-1)和微流(1-20 ul min-1)层析分离，以便在临床研究的背景下对有限的样本量进行深入分析，并对较大的患者/先证者队列进行高通量分析。

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.