Characterization of the Time-Domain Isotopic Beat Patterns of Monoclonal Antibodies in Fourier Transform Mass Spectrometry,ACS Environmental Au

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Characterization of the Time-Domain Isotopic Beat Patterns of Monoclonal Antibodies in Fourier Transform Mass Spectrometry
ACS Environmental Au ( IF 6.7 ) Pub Date : 2022-05-31 , DOI: 10.1021/jasms.1c00336
Konstantin O. Nagornov ₁ , Anton N. Kozhinov ₁ , Natalia Gasilova ₂ , Laure Menin ₂ , Yury O. Tsybin ₁

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The time-domain transients in the Fourier transform mass spectrometry (FTMS) analysis of monoclonal antibodies (mAbs) are known to exhibit characteristic isotopic beat patterns. These patterns are defined by the isotopic distributions of all gaseous mAb ions present in the FTMS mass analyzer, originating from single or multiple charge states, and from single or multiple proteoforms. For an isolated charge state of a single proteoform, the mAb isotopic beat pattern resembles narrow splashes of signal amplitude (beats), spaced periodically in the time-domain transient, with broad (often exceeding 1 s) “valleys” between them. Here, we reinforce the importance of isotopic beat patterns for the accurate interpretation and presentation of FTMS data in the analysis of mAbs and other large biopolymers. An updated, mAb-grade version of the transient-mediated FTMS data simulation and visualization tool, FTMS Simulator is introduced and benchmarked. We then apply this tool to evaluate the charge-state dependent characteristics of isotopic beats in mAbs analyses with modern models of Orbitrap and ion cyclotron resonance (ICR) FTMS instruments, including detection of higher-order harmonics. We demonstrate the impact of the isotopic beat patterns on the analytical characteristics of the resulting mass spectra of individual and overlapping mAb proteoforms. The results reported here detail highly nonlinear dependences of resolution and signal-to-noise ratio on the time-domain transient period, absorption or magnitude mode spectra representation, and apodization functions. The provided description and the demonstrated ability to routinely conduct accurate simulations of FTMS data for large biopolymers should aid the end-users of Orbitrap and ICR FTMS instruments in the analysis of mAbs and other biopolymers, including viruses.

中文翻译：

傅里叶变换质谱法中单克隆抗体时域同位素跳动模式的表征

已知单克隆抗体 (mAb) 的傅里叶变换质谱 (FTMS) 分析中的时域瞬变表现出特征同位素搏动模式。这些模式由 FTMS 质量分析器中存在的所有气态 mAb 离子的同位素分布定义，源自单个或多个电荷状态，以及来自单个或多个 proteoforms。对于单个蛋白质形式的孤立电荷状态，mAb 同位素拍动模式类似于信号幅度（拍）的窄飞溅，在时域瞬态中周期性地间隔，它们之间具有宽（通常超过 1 秒）“谷”。在这里，我们强调同位素搏动模式对于在 mAb 和其他大型生物聚合物分析中准确解释和呈现 FTMS 数据的重要性。一个更新的，mAb 级版本的瞬态介导 FTMS 数据模拟和可视化工具，FTMS Simulator 被引入和基准测试。然后，我们应用这个工具来评估 mAbs 分析中同位素跳动的电荷状态相关特征，使用现代模型的 Orbitrap 和离子回旋共振 (ICR) FTMS 仪器，包括检测高次谐波。我们展示了同位素节拍模式对单个和重叠 mAb 蛋白型的质谱分析特征的影响。此处报告的结果详细说明了分辨率和信噪比对时域瞬态周期、吸收或幅度模式光谱表示以及变迹函数的高度非线性依赖性。

更新日期：2022-05-31

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