Mass spectrometric system with single-cell-sensitivity for quantitative proteomics

要求为核心设施提供一套系统，用于分析通常存在于单细胞中的极小样本量的蛋白质组。该系统包括一个纳米-超高压液相色谱（nano-UPLC），包括一个自动进样器，一个高分辨率串联质谱（hr-MSMS），一个电喷雾电离（ESI）源，一个离子迁移模块以及必要的软件模块。对于通过激光烧蚀（3- ti - lab）进行的三维组织微采样的非常小的样本量的分析，也需要非常高的灵敏度。作为废弃纳米uplc - msms的替代购买，所要求的系统还将用于分析µg范围内的更大样本量的差异定量自下而上蛋白质组学，包括无标签和基于标签的，例如SILAC或16plx - tmt定量方法，用于分析蛋白质形态的定量自上而下蛋白质组学，以及不同类型的定量糖组学。除了在核心设施中运行该系统（容量利用率为80 - 90%）外，Hartmut schl<e:1>教授小组还将使用该系统开发和改进3- mit - lab技术以及分析proteoforms。由Klaus Pantel教授领导的肿瘤生物学研究所将分析循环肿瘤细胞（ctc）的蛋白质组。今后两组仪器系统的容量利用率将不超过每组仪器系统的10%。

A system is requested for a Core Facility for analysis of proteomes of very small sample amounts, which are typically present in single cells. The system consists of a nano-ultra-high-pressure liquid chromatography (nano-UPLC), including an autosampler, and a high-resolution tandem mass spectrometer (hr-MSMS) with an electrospray-ionization (ESI) source an ion mobility module as well as necessary software modules. A very high sensitivity is required also for analysis of very small sample amounts derived from the 3-dimensional tissue µ-sampling via laser ablation (3-MiTi-LAb). As replacement purchase of a discarded nano-UPLC-MSMS the requested system will also be used for analysis of larger sample amounts in the µg range for differential quantitative bottom-up proteomics, including label-free and label-based, e.g. SILAC or 16plex-TMT quantification methods, quantitative top-down proteomics for analysis of proteoforms, as well as for different types quantitative glycomics. Beside the operation of the system in the Core Facility, comprising 80 – 90 % capacity utilization, the group of Prof. Hartmut Schlüter will use the system for developing and improving the 3-MiTi-LAb technology as well as the analysis of proteoforms. The Institute of Tumor Biology, headed by Prof. Klaus Pantel, will analyze the proteomes of circulating tumor cells (CTCs). Both groups in the future will not exceed 10 percent capacity utilization of the instrument system per each group.