Coupling High-Field Asymmetric Waveform Ion Mobility Spectrometry with Capillary Zone Electrophoresis-Tandem Mass Spectrometry for Top-Down Proteomics,Analytical Chemistry

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Coupling High-Field Asymmetric Waveform Ion Mobility Spectrometry with Capillary Zone Electrophoresis-Tandem Mass Spectrometry for Top-Down Proteomics
Analytical Chemistry ( IF 6.7 ) Pub Date : 2023-05-30 , DOI: 10.1021/acs.analchem.3c00551
Tian Xu ₁ , Qianjie Wang ₁ , Qianyi Wang ₁ , Liangliang Sun ₁

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Capillary zone electrophoresis-tandem mass spectrometry (CZE-MS/MS) has emerged as an essential technique for top-down proteomics (TDP), providing superior separation efficiency and high detection sensitivity for proteoform analysis. Here, we aimed to further enhance the performance of CZE-MS/MS for TDP via coupling online gas-phase proteoform fractionation using high-field asymmetric waveform ion mobility spectrometry (FAIMS). When the compensation voltage (CV) of FAIMS was changed from −50 to 30 V, the median mass of identified proteoforms increased from less than 10 kDa to about 30 kDa, suggesting that FAIMS can efficiently fractionate proteoforms by their size. CZE-FAIMS-MS/MS boosted the number of proteoform identifications from a yeast sample by nearly 3-fold relative to CZE-MS/MS alone. It particularly benefited the identification of relatively large proteoforms, improving the number of proteoforms in a mass range of 20–45 kDa by 6-fold compared to CZE-MS/MS alone. FAIMS fractionation gained nearly 20-fold better signal-to-noise ratios of randomly selected proteoforms than no FAIMS. We expect that CZE-FAIMS-MS/MS will be a useful tool for further advancing the sensitivity and coverage of TDP. This work shows the first example of coupling CE with ion mobility spectrometry (IMS) for TDP.

中文翻译：

高场不对称波形离子淌度谱与毛细管区带电泳-串联质谱联用，实现自上而下的蛋白质组学

毛细管区带电泳-串联质谱 (CZE-MS/MS) 已成为自上而下蛋白质组学 (TDP) 的一项重要技术，为蛋白质型分析提供卓越的分离效率和高检测灵敏度。在这里，我们的目标是通过使用高场不对称波形离子迁移谱 (FAIMS) 耦合在线气相蛋白质形式分馏，进一步提高 TDP 的 CZE-MS/MS 性能。当 FAIMS 的补偿电压 (CV) 从 -50 V 变为 30 V 时，所鉴定的蛋白质型的中值质量从小于 10 kDa 增加到约 30 kDa，这表明 FAIMS 可以根据蛋白质型的大小有效地分级。与单独使用 CZE-MS/MS 相比，CZE-FAIMS-MS/MS 将酵母样品中蛋白质型的鉴定数量提高了近 3 倍。它特别有利于鉴定相对较大的蛋白质形式，与单独的 CZE-MS/MS 相比，质量范围为 20-45 kDa 的蛋白质形式数量提高了 6 倍。 FAIMS 分级分离随机选择的蛋白质形式的信噪比比没有 FAIMS 提高了近 20 倍。我们预计 CZE-FAIMS-MS/MS 将成为进一步提高 TDP 灵敏度和覆盖范围的有用工具。这项工作展示了将 CE 与离子迁移谱 (IMS) 耦合用于 TDP 的第一个示例。

更新日期：2023-05-30

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