Ultrahigh-Throughput Intact Protein Analysis with Acoustic Ejection Mass Spectrometry
Ultrahigh-Throughput Intact Protein Analysis with Acoustic Ejection Mass Spectrometry
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DOI：
10.1021/jasms.2c00276
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发表时间：
2022-12-05
影响因子：
3.2
通讯作者：
McLaren, David G.
中科院分区：
文献类型：
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作者：
Zacharias, Adway O.;Liu, Chang;McLaren, David G.
随着药物发现越来越需要分析大量共价修饰剂和蛋白质治疗剂，对高通量完整蛋白质分析的需求一直在上升。液相色谱-质谱（LC MS）是迄今为止用于生物制药行业中表征蛋白质的主要分析工具。然而，LC-MS的速度阻止了大规模样品组的分析（一天内>1000）。声喷射质谱（AEMS）是近年来发展起来的一种基于电喷雾离子化（ESI）-MS的分析平台，具有分析速度快、数据质量高等优点。自引入以来，该技术已应用于许多领域，主要集中在高通量药物发现和开发中的小分子分析。在这里，我们探讨AEMS的应用程序，以高通量完整的蛋白质分析的蛋白质范围内的分子量从17至150 kDa的原型高分辨率四极杆飞行时间（HR QTOF）为基础的AEMS系统。在该平台上获得的数据质量与LC-MS相当，而分析速度显着提高到每个样品一秒。这种超高通量完整蛋白质分析平台具有广泛应用于药物发现的潜力。
The need for high-throughput intact protein analysis has been rising as drug discovery increasingly requires the analysis of large sets of covalent modifiers and protein therapeutics. Liquid chromatography-mass spectrometry (LC MS) is the primary analytical tool used to date to characterize proteins within the biopharmaceutical industry. However, the speed of LC-MS prevents the analysis of large-scale sample sets (>1000 within a day). Acoustic ejection mass spectrometry (AEMS) has recently been established as an electrospray ionization (ESI)-MS based platform with both fast analytical throughput and high data quality. Since its introduction, this technology has been applied in numerous fields with a primary focus on small-molecule analysis in high-throughput drug discovery and development. Here we explore the application of AEMS to high-throughput intact protein analysis for proteins ranging in molecular weight from 17 to 150 kDa on a prototype high-resolution quadrupole time-of flight (HR QTOF) based AEMS system. Data quality obtained on this platform is comparable to LC-MS, while the analysis speed is significantly improved to one-second-per-sample. This ultrahigh-throughput intact protein analysis platform has the potential to be used broadly in drug discovery.