Assessment of the repeatability and reproducibility of hydrogen/deuterium exchange mass spectrometry measurements
Assessment of the repeatability and reproducibility of hydrogen/deuterium exchange mass spectrometry measurements
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DOI：
10.1002/rcm.3794
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发表时间：
2008-12-01
影响因子：
2
通讯作者：
O'Connor, Gavin
中科院分区：
文献类型：
--
作者：
Burkitt, William;O'Connor, Gavin
使用能够进行溶剂操作的XYZ机器人自动进样器和4.7 T傅里叶变换离子回旋共振（FT-ICR）质谱仪构建了进行自动氢/氘交换质谱测量的系统。该系统包括的功能，如一个'双柱'高效液相色谱（HPLC）设置的第一次演示，和一个新的消化策略。在蛋白质氢/氘交换测量的重复性和再现性方面，在2个月的时间内评估了系统的性能。对几个参数的氢交换测量的灵敏度进行了评估，这使得它们对重现性的影响进行了讨论。评估的参数包括HPLC柱和开关阀的温度、淬灭溶液的温度、移动的相的pH值、淬灭溶液的pH值、移动的相中使用的酸和使用的分析柱。版权所有（C）2008约翰威利父子有限公司
A system to perform automated hydrogen/deuterium exchange mass spectrometry measurements was constructed using an XYZ robotic autosampler that was capable of performing solvent manipulations and a 4.7 T Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometer. The system included features such as the first demonstration of a 'dual column' high-performance liquid chromatography (HPLC) setup, and a novel digestion strategy. The performance of the system, in terms of the repeatability and reproducibility of the measurement of protein hydrogen/deuterium exchange, was assessed over a 2-month period. The sensitivity of the measurement of hydrogen exchange towards several parameters was assessed, which allowed their impact on the reproducibility to be discussed. The parameters assessed were the temperature of the HPLC columns and switching valves, the temperature of the quench solutions, the pH of the mobile phase, the pH of the quenched solution, the acid used in the mobile phase and the analytical column used. Copyright (C) 2008 John Wiley & Sons, Ltd.