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

需要一个用于分析通常存在于单细胞中的极小样本量的蛋白质组的核心设备的系统。该系统由包括自动进样器的纳米超高压液相色谱(Nano-UPLC)和带有电喷雾电离(ESI)源的高分辨率串联质谱仪(hr-MSMS)、离子迁移模块以及必要的软件模块组成。对于通过激光消融的三维组织采样(3-MITI-LAB)获得的非常少量的样本量的分析，也需要非常高的灵敏度。作为对废弃纳米UPLC-MSMS的替换购买，所请求的系统还将用于分析微克范围内的较大样本量，用于自下而上的差异定量蛋白质组学，包括无标记和基于标记的，例如SILAC或16plex-TMT定量方法，用于蛋白质形式分析的自顶向下的定量蛋白质组学，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，包括80%-90%的产能利用率外，Hartmut Schlüter教授小组还将使用该系统开发和改进3-MITI-LAB技术以及蛋白质组分析。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(CTCs)的蛋白质组。未来两个组每组的仪器系统容量利用率不超过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.