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

图宾根大学校园对高端蛋白质组分析的需求在过去几年中显着增加。这主要是由于基于质谱的蛋白质组分析技术已经取得了显着进步，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。目前的仪器已经运行了8年多，并且已经大量使用，由于维护而导致的停机时间在过去两年中显著增加。这导致在我们的核心设施中等待分析样品的队列超过两个月，不幸的是，由于蒂宾根医学园区的需求不断增加和项目不断增加，我们不得不拒绝样品测量。此外，该技术已经过时，不能满足目前对小而复杂样品进行深入蛋白质组学分析的需求，因此需要用最先进的仪器取代。因此，我们要求用新仪器更换这台质谱仪。随着我们越来越多的项目采用非常有限的样本量，例如内源性蛋白质复合物、原代组织以及基于诱导多能干细胞的来源，其中需要具有高扫描速率和超高分辨率的高灵敏度设备。此外，为了提高通量和重现性，新仪器必须允许数据独立采集，并且为了验证目的和检测复杂样品中的特定蛋白质，必须对特定肽进行有针对性的分析。特别是，有针对性的分析是必不可少的转化研究，以及在大样本数量的验证。此外，为了分离复杂的混合物，需要将超高效液相色谱系统连接到质谱仪，这允许以高分辨率和纳米流速可再现地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。

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 Moreover, the technology is outdated, not satisfying current needs for in-depth proteomic analysis of small and complex samples and therefore needs to be replaced with a state-of-the-art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.