LC-MS/MS system with high-resolution mass spectrometer and nano-UHPLC - 1/2

配备高分辨率质谱仪和纳米 UHPLC 的 LC-MS/MS 系统 - 1/2

• 批准号: 527318136
• 金额: --
• 依托单位: Eberhard Karls Universität Tübingen
• 依托单位国家: 德国
• 项目类别: Major Research Instrumentation
• 财政年份: 2023
• 资助国家: 德国
• 起止时间: 2022-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/527318136?language=en
• 关键词: LC; MS; system; resolution; mass

Proteomic analysis in translational research has recently gained a lot of momentum. New technologies for sample processing as well as mass spectrometric analysis now enable us to establish novel workflows for a variety of biomedical research applications and even diagnostics. The current instrumentation has been in use for more than 12 years continuously, and the maintenance-related downtime has increased significantly over the past four years. Additionally, it is not competitive any longer, mainly due to slow acquisition and due to the lack of possibilities to perform novel analysis workflows like data-independent mass spectrometry or similar approaches. Therefore, it needs to be replaced with a state-of-the-art instrument. To replace this machine, we request a new instrument. It is planned to be used for various proteomic applications including the analysis of formalin-fixed and paraffine-embedded tissue samples where only limited material is available. Also, the analysis of small samples for relatively small tissue or organoid samples including pull-downs for protein interaction analysis via tagged fusion protein capture and antibody-based pull-downs is increasing and the analysis of body fluids like tear fluid and perilymph needs a highly sensitive and reliable Liquid Chromatography-Mass Spectrometry system with respect to sensitivity, throughput and analytical depth. This requires a device with high scan rates and ultra-high resolution for large sample throughput of very complex as well as very scarce samples. Further, to increase the throughput and reproducibility, the new instrument needs to enable high resolution and high-speed data-independent acquisition and, for validation purposes and to detect specific proteins in complex samples, targeted analysis of specific peptides. Additionally, for separation of the complex mixtures, a ultra-high pressure liquid chromatography system needs to be connected to the mass spectrometer which allows high resolution separation of complex peptide mixtures at nano flow rates. The set-up we are requesting is a combination of a nano ultra-high performance liquid chromatography system, coupled with an ion-mobility unit connected to an ultra-high resolution mass spectrometry machine. This combination is a perfect setup to achieve a precise retention time, limit sample dispersion and system carryover, reduce spectral noise background and deliver sensitive and accurate identification and quantification of peptides at a high resolution.

翻译研究中的蛋白质组学分析最近获得了很大的动力。新的样品处理和质谱分析技术使我们能够为各种生物医学研究应用甚至诊断建立新的工作流程。目前的仪器已经连续使用了12年多，在过去的四年中，与维护相关的停机时间显著增加。此外，它不再具有竞争力，主要是由于采集速度慢，以及缺乏执行新的分析工作流程（如数据独立质谱法或类似方法）的可能性。因此，需要用最先进的仪器取代它。为了更换这台机器，我们要求一台新仪器。它计划用于各种蛋白质组学应用，包括福尔马林固定和石蜡包埋的组织样本的分析，其中只有有限的材料。此外，对相对小的组织或类器官样品的小样品的分析，包括通过标记的融合蛋白捕获和基于抗体的下拉进行蛋白质相互作用分析的下拉正在增加，并且体液如泪液和外淋巴的分析需要在灵敏度、通量和分析深度方面高度灵敏和可靠的液相色谱-质谱系统。这就需要一种具有高扫描速率和超高分辨率的设备，以实现非常复杂以及非常稀缺的样品的大样品吞吐量。此外，为了提高通量和再现性，新仪器需要实现高分辨率和高速数据独立采集，并用于验证目的和检测复杂样品中的特定蛋白质，对特定肽进行靶向分析。此外，为了分离复杂的混合物，需要将超高压液相色谱系统连接到质谱仪，其允许以纳米流速高分辨率分离复杂的肽混合物。我们要求的设置是一个纳米超高效液相色谱系统的组合，加上连接到超高分辨率质谱仪的离子迁移率单位。这种组合是一种完美的设置，可实现精确的保留时间，限制样品分散和系统携带，降低光谱噪声背景，并以高分辨率提供灵敏准确的肽鉴定和定量。