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

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

• 批准号: 527318363
• 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: 2024
• 资助国家: 德国
• 起止时间: 2023-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/527318363?language=en
• 关键词: LC; MS; system; resolution; mass

The demand for high-end proteome analysis on the university campus Tübingen has increased significantly over the past few years. This is mainly due to the fact that the technology of mass spectrometry-based proteome analysis has dramatically advanced and now offers a great variety of analytical options for many areas of bio-medical research. The current instrumentation has been in operation for more than 8 years and has been heavily used, the downtime due to maintenance has significantly increased over the last two years. This has resulted in a waiting queue of more than two months for samples to be analysed in our core facility, and unfortunately, we had to reject sample measurements due to increasing demand and rising projects for the medical campus of Tübingen. Moreover, the technology is outdated, not satisfying current needs for in-depth proteomic analysis of small and complex samples and therefore needs to be replaced with a state-of-the-art instrument. Therefore, we request to replace this mass spectrometer, with a new instrument. As we approach an increasing number of projects with very limited sample sizes derived of for example endogenous protein complexes, primary tissue as well as of induced pluripotent stem cell-based origin, where a highly sensitive device with high scan rates and ultra-high resolution is needed. Furthermore, to increase both the throughput and reproducibility, the new instrument has to allow data-independent acquisition and, for validation purposes and to detect specific proteins in complex samples, targeted analysis of specific peptides. In particular, targeted analysis is essential for translational research as well as for validation in large sample numbers. Additionally, for separation of the complex mixtures, an ultra-high performance liquid chromatography system needs to be connected to the mass spectrometer which allows a reproducible separation of complex peptide mixtures at high resolution and nano flow rates. The set-up we are requesting is a combination of a nano ultra-high performance liquid chromatography system, coupled with a 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.

在过去几年中，对图宾根大学校园对高端蛋白质组分析的需求已大大增加。这主要是由于以下事实：基于质谱的蛋白质组分析的技术已经大大提高，现在为生物医学研究的许多领域提供了各种各样的分析选择。当前的仪器已经运行了8年以上，并且已经大量使用了，在过去的两年中，由于维护而导致的停机时间大大增加。这导致了在我们的核心设施中分析样品超过两个月的等待队列，不幸的是，由于需求的增加和Tübingen医疗校园的不断增长，我们不得不拒绝样品测量结果。此外，该技术已过时，无法满足当前对小型和复杂样品的深入蛋白质组学分析的需求，因此需要用最先进的仪器代替。因此，我们要求用新仪器代替该质谱仪。当我们接近越来越多的项目时，具有非常有限的样本量得出的项目，例如内源性蛋白质复合物，原代组织以及诱导的多能干细胞基于基于干细胞的起源，其中需要具有高扫描速率和超高分辨率的高度敏感的设备。此外，为了增加吞吐量和可重复性，新仪器必须允许独立于数据的获取，并出于验证目的并检测复杂样品中的特定蛋白质，对特定肽的靶向分析。特别是，有针对性的分析对于转化研究以及大量样本数量的验证至关重要。此外，对于复合混合物的分离，需要将超高的性能液相色谱系统连接到质谱仪，该系统允许以高分辨率和纳米流速以可重现的复合肽混合物可重复分离。我们要求的设置是纳米超高性能液相色谱系统的组合，并与连接到超高分辨率质谱机的离子摩托车单元相结合。这种组合是实现精确保留时间，限制样品分散和系统结转，减少光谱噪声背景并提供高分辨率肽的敏感，准确的识别和定量的理想设置。