Hybrid-Massenspektrometer mit einem Gradienten Fokussierungssystem für die Flüssigkeitschromatographie

近年来，基于质谱的蛋白质组学日益成为许多生物学和医学问题的关键技术。应用质谱仪具有灵敏度高、测序速度快、质量精度高等特点，可采用碰撞诱导离解（CID）、高能c阱离解（HCD）和电子转移离解（ETD）等多种裂解技术。使用这种高分辨率的四极离子阱质谱仪，可以深入分析复杂的生物样品。除标准配置的高效液相色谱装置外，还应用了另一套液相色谱系统，该系统专为高样品通量和短梯度项目而设计。这个额外的色谱单位将在科隆建立系统医学方面发挥重要作用。质谱仪还允许通过ETD对完整的蛋白质进行碎片化。到目前为止，这种方法在科隆大学还不可行。不同类型的片段（CID， ETD）的组合也提供了一个最佳的表征交联肽的质谱-可切割的交联分子。与生物化学系密切合作，蛋白质复合物的结构将使用交联质谱（XL-MS）和低温电子显微镜的组合进行研究。CID碎片将使用离子阱质谱仪（LTQ-Orbitrap Discovery， 2008年购买）进行，该质谱仪已不再是最先进的。总的来说，提议的质谱仪将增加该设施的容量，并显着扩大质谱仪方法的范围。

In recent years, mass spectrometry-based proteomics has increasingly become the key technology for many biological and medical questions. The applied mass spectrometer possesses high sensitivity, fast sequencing speed, high mass accuracy and diverse fragmentation techniques such as collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD) and electron-transfer dissociation (ETD). Using this high-resolution quadrupole ion-trap mass spectrometer, complex biological samples can be analyzed in depth. In addition to the standard configuration with a high-performance liquid chromatography device, a further liquid chromatography system was applied for, which was designed for projects with high sample throughput and short gradients. This additional chromatography unit will play an important role in the establishment of systems medicine in Cologne. The mass spectrometer also allows the fragmentation of intact proteins via ETD. So far, this method is not feasible at the University of Cologne. The combination of different types of fragmentation (CID, ETD) also offers an optimal representation of cross-linked peptides by mass spectrometry-cleavable cross-linker molecules. In close cooperation with the biochemistry department, the structure of protein complexes will be investigated using a combination of cross-linking mass spectrometry (XL-MS) and cryo-electron microscopy. CID fragmentation will be performed with an ion trap mass spectrometer (LTQ-Orbitrap Discovery; purchased 2008), which is no longer state of the art. Overall, the proposed mass spectrometer will increase the capacity of the facility and significantly expand the range of mass spectrometric methods.