Analytical Aspects of Hydrogen Exchange Mass Spectrometry.
Analytical Aspects of Hydrogen Exchange Mass Spectrometry.
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DOI：
10.1146/annurev-anchem-062011-143113
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Wales TE
中科院分区：
文献类型：
--
作者：
Engen JR;Wales TE
回顾了通过质谱法测量氢交换的分析方面。描述了氢交换中分析选择性的性质，然后回顾了在限制性交换淬灭条件下完成裂解、分离和质谱测量所需的分析工具。与依赖于峰强度或面积测量的分析定量相反，氢交换质谱中的定量取决于测量相对于未氘化或氘化对照的质量变化，从而产生介于零和可掺入的最大氘量之间的值。可靠的定量取决于实验保真度，为了实现高测量重现性，在样品制备和分析过程中必须控制大量实验变量。该方法还报告样本的重要定性方面，包括构象异质性和群体动态。
The analytical aspects of measuring hydrogen exchange by mass spectrometry are reviewed. The nature of analytical selectivity in hydrogen exchange is described followed by review of the analytical tools required to accomplish fragmentation, separation, and the mass spectrometry measurements under restrictive exchange quench conditions. In contrast to analytical quantitation that relies on measurements of peak intensity or area, quantitation in hydrogen exchange mass spectrometry depends on measuring a mass change with respect to an undeuterated or deuterated control, resulting in a value between zero and the maximum amount of deuterium that could be incorporated. Reliable quantitation is a function of experimental fidelity and to achieve high measurement reproducibility, a large number of experimental variables must be controlled during sample preparation and analysis. The method also reports on important qualitative aspects of the sample, including conformational heterogeneity and population dynamics.