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

用于定量蛋白质组学的具有单细胞灵敏度的质谱系统

• 批准号: 518551069
• 金额: --
• 依托单位: Universität Hamburg
• 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/518551069?language=en
• 关键词: Mass; spectrometric; system; single; cell

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.

要求系统建立一个核心设施，以分析非常小的样品量，通常存在于单个单元格中。该系统由纳米 - 粉状压力液相色谱（Nano-UPLC）组成，包括自动采样器，以及具有电喷雾离子化（ESI）源的高分辨率串联质谱仪（HR-MSMS），一个离子迁移率模块以及必要的软件模块。还需要非常高的灵敏度来分析通过激光消融（3-MITI-LAB）源自3维组织的样品量。随着替换丢弃的纳米 - UPLC-MSM的替换，所请求的系统还将用于分析较大的样品量，以用于差异定量的自下而上蛋白质组学，包括基于标签和标签的差异，例如。 SILAC或16PLEX-TMT定量方法，用于分析蛋白质成型的定量自上而下的蛋白质组学以及不同类型的定量糖核酸。除了系统在核心设施中的运行外，完成80 - 90％的容量利用率外，HartmutSchlüter教授将使用该系统来开发和改进3 MITI-LAB技术以及蛋白质类型的分析。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（CTC）的蛋白质体。将来，每个组的仪器系统的容量利用率都不会超过10％。