Nägele Edgar, Vollmer Martin
Agilent Technologies R&D and Marketing GmbH & Co. KG, Hewlett-Packard-Str. 8, 76337 Waldbronn, Germany.
Rapid Commun Mass Spectrom. 2004;18(24):3008-14. doi: 10.1002/rcm.1723.
The major obstacle in the use of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) instruments in the analysis of complex proteome samples is the lack of a direct coupling of a highly resolving separation technique with the mass spectrometer itself. To overcome this drawback, a spotting device for capillary and nanoflow liquid chromatography (LC) with a special liquid deposition principle for lowest volumes was developed. The instrument is able to perform MALDI spotting in real time in order to deposit the LC run on the MALDI plate, and therefore couples the high resolution power of nano-RP-HPLC separation directly with MALDI-MS. This work describes the development and optimization of a method for spotting with online matrix addition, and illustrates its use in the analysis of a complex proteome sample.
在使用基质辅助激光解吸/电离质谱（MALDI-MS）仪器分析复杂蛋白质组样品时，主要障碍是缺乏高分辨率分离技术与质谱仪本身的直接耦合。为克服这一缺点，开发了一种用于毛细管和纳流液相色谱（LC）的点样装置，其具有特殊的液体沉积原理以实现最低体积。该仪器能够实时进行MALDI点样，以便将LC运行的产物沉积在MALDI板上，从而将纳米反相高效液相色谱（nano-RP-HPLC）分离的高分辨率能力直接与MALDI-MS耦合。本文描述了一种在线添加基质进行点样方法的开发和优化，并说明了其在分析复杂蛋白质组样品中的应用。
