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

我们正在申请高分辨率电喷雾电离串联质谱仪的资金，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，并与纳米流和微流色谱系统相结合。该仪器将安装并用于Göttingen大学医学中心Göttingen （UMG）的核心设施蛋白质组学，这是一个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的蛋白质组学分析。该设施成立于2012年，配备了第一代质谱仪器。这里描述的申请仪器将取代2012年安装的老化质谱仪之一。申请的仪器必须支持以下研究方法：1)数据独立获取质谱法（DIA-MS），用于细胞培养物、体液和组织样品的中至高通量无标签分析2)数据依赖获取质谱法（DDA-MS），使用等压重同位素标签（例如串联质量标签），TMT)用于深入定量分析全局蛋白表达和翻译后修饰（PTM）状态3)dda - ms和DIA-MS分析化学交联蛋白样品，用于全局分析蛋白/蛋白相互作用位点4)dda - ms和DIA-MS分析蛋白质亲和纯化（AP）和蛋白质接近标记实验，以确定蛋白/蛋白相互作用要求包括离子迁移分离阶段，例如在质量的界面区域光谱仪为分析高度复杂的样品提供额外的选择性水平，并且能够操作纳流（100-350 nl min-1）和微流（1-20 ul min-1）色谱分离，以便对有限的样本量进行深入分析，以及在临床研究背景下对较大的患者/先证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.