High-resolution ultra-high mass spectrometry: increasing the m/z range of protein analysis.
High-resolution ultra-high mass spectrometry: increasing the m/z range of protein analysis.
复制标题
高分辨率超高分辨率质谱：增加蛋白质分析的 m/z 范围。
DOI：
10.1002/pmic.201270136
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发表时间：
2012
期刊：
影响因子：
3.4
通讯作者：
Reilly,PeterTA
中科院分区：
文献类型：
--
作者：
Chen,Huijuan;Lee,Jeonghoon;Reilly,PeterTA
最近已经证明了跨越整个范围的完整单电荷蛋白质的高分辨率 TOF 质量分析的原理证明。通过内部校准，复杂分布中单个蛋白质的同位素分布中心可以定义在 0.5 Da 以内，甚至更好，最高可达 200 kDa。随着该技术成为主流，这一成就将对未来几年的常规蛋白质分析过程产生巨大影响。超高质量范围 (m/z> 20 000) 中实现高分辨率的最大障碍是膨胀引起的动能 (KE) 及其扩散。该问题的解决方案是将离子捕获在缓冲气体中，以便离子的运动可以完全由施加的场定义。如果这可以在不依赖质量的情况下完成，那么任何离子，无论大小，都可以进行高分辨率的质量分析。本文讨论了我们用来在真空中捕获大气中的单电荷蛋白质进入质量分析器之前的点的方法，以完全消除膨胀引起的 KE。然后，我们使用数字创建的四极波形将离子注入质量分析仪中具有受控 KE 量的准直插头中。捕获离子以消除膨胀引起的 KE，然后以电动方式操纵离子是在任何 m/z 值下进行高分辨率质量分析的关键步骤。将讨论该技术的影响。
The proof of principle for high‐resolution TOF mass analysis spanning the entire range of intact singly charged proteins has recently been demonstrated. The centers of the isotope distributions of individual proteins in a complex distribution can be defined to within 0.5 Da or better up to 200 kDa with internal calibration. This achievement will have an enormous effect on the process of routine protein analysis over the next few years as the technology mainstreams. The greatest obstacle to high‐resolution in the ultra‐high mass range (m/z> 20 000) is the expansion‐induced kinetic energy (KE) and its spread. The solution to this problem is to trap the ions in a buffer gas so that the motion of the ions can be completely defined by the applied fields. If this can accomplished without mass dependence, then any ion, regardless of size, can be mass analyzed with high resolution. This article discusses the methodology that we used to capture atmosphere sampled singly charged proteins in vacuum at a point just before they enter the mass analyzer to completely eliminate the expansion‐induced KE. We then used digitally created quadrupole waveforms to inject the ions into the mass analyzer in a well‐collimated plug with a controlled amount of KE. Trapping the ions to remove the expansion‐induced KE and then electrodynamically manipulating the ions are the key steps to high‐resolution mass analysis at any value ofm/z. The impact of this technology will be discussed.