LC/MS at the whole protein level: Studies of biomolecular structure and interactions using native LC/MS and cross-path reactive chromatography (XP-RC) MS
LC/MS at the whole protein level: Studies of biomolecular structure and interactions using native LC/MS and cross-path reactive chromatography (XP-RC) MS
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DOI：
10.1016/j.ymeth.2018.04.019
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发表时间：
2018-07-15
期刊：
影响因子：
4.8
通讯作者：
Lipatnikov, Andrei N.
中科院分区：
文献类型：
--
作者：
Kaltashov, Igor A.;Pawlowski, Jake W.;Lipatnikov, Andrei N.
将液相色谱（LC）与电喷雾离子化（ESI）连接以实现在线MS检测最初使用反相LC来实现，在过去三十年中，反相LC仍然是用于LC/MS和LC/MS/MS研究蛋白质结构的默认色谱类型。相比之下，其他类型的LC作为ESI MS前端的优点，特别是那些允许生物聚合物高阶结构在整个分离过程中得到保留的优点，直到最近才得到相对较少的赞赏。然而，在过去的几年里，在生物分析和生物物理界，对所谓的“天然”（“非变性”可能是一个更合适的形容词）LC/MS和LC/MS/MS分析的兴趣急剧增加。本文综述了这一领域的最新进展，重点是尺寸排阻和离子交换色谱作为LC/MS蛋白质表征的前端平台。还讨论了在天然LC/MS分析中整合化学反应所带来的好处，包括气相中的离子化学（例如，用于表征高度非均相生物聚合物的有限电荷减少）和溶液相反应（使用最近引入的技术交叉路径反应色谱法）。
Interfacing liquid chromatography (LC) with electrospray ionization (ESI) to enable on-line MS detection had been initially implemented using reversed phase LC, which in the past three decades remained the default type of chromatography used for LC/MS and LC/MS/MS studies of protein structure. In contrast, the advantages of other types of LC as front-ends for ESI MS, particularly those that allow biopolymer higher order structure to be preserved throughout the separation process, enjoyed relatively little appreciation until recently. However, the past few years witnessed a dramatic surge of interest in the so-called "native" (with "non-denaturing" being perhaps a more appropriate adjective) LC/MS and LC/MS/MS analyses within the bioanalytical and biophysical communities. This review focuses on recent advances in this field, with an emphasis on size exclusion and ion exchange chromatography as front-end platforms for protein characterization by LC/MS. Also discussed are the benefits provided by the integration of chemical reactions in the native LC/MS analyses, including both ion chemistry in the gas phase (e.g., limited charge reduction for characterization of highly heterogeneous biopolymers) and solution-phase reactions (using the recently introduced technique cross-path reactive chromatography).