A tandem mass spectrometer for improved transmission and analysis of large macromolecular assemblies
A tandem mass spectrometer for improved transmission and analysis of large macromolecular assemblies
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DOI：
10.1021/ac0110552
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发表时间：
2002-03-15
影响因子：
7.4
通讯作者：
Robinson, CV
中科院分区：
文献类型：
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作者：
Sobott, F;Hernández, H;Robinson, CV
我们报告的设计和第一个应用程序的串联质谱仪（四极杆飞行时间质谱仪）优化传输和分析的大分子组件。已经发现，仔细控制仪器的不同泵送阶段中的压力梯度对于检测大分子颗粒是必不可少的。然而，这样的组件难以通过串联MS方法分析，因为它们产生高于m/z 3000-4000的信号，这是商业四极杆的极限。通过将四极杆的频率降低到300 kHz并将其用作窄带质量过滤器，我们表明可以在窄至22 m/z单位的窗口中从m/z 22 000处的单峰中分离离子。使用碘化铯团簇信号，我们表明，在飞行时间（TOF）分析仪的质量范围扩展到超过m/z 90 000，在理论上超过m/z 150 000。我们还表明，该仪器的分辨率大于3000 m/z 44 500。串联质谱的能力，说明从异源寡聚体组装之间形成的四聚体甲状腺素运载蛋白，甲状腺素，视黄醇结合蛋白，和视黄醇分离组件。在四极杆中m/z 5340处的单个电荷状态的隔离和随后在充气碰撞室中的碰撞诱导解离（CID）导致从单个亚基和亚复合物形成离子，通过TOF分析仪中的质量和电荷来识别。
We report the design and first applications of a tandem mass spectrometer (a quadrupole time-of-flight mass spectrometer) optimized for the transmission and analysis of large macromolecular assemblies. Careful control of the pressure gradient in the different pumping stages of the instrument has been found to be essential for the detection of macromolecular particles. Such assemblies are, however, difficult to analyze by tandem-MS approaches, because they give rise to signals above m/z 3000-4000, the limit for commercial quadrupoles. By reducing the frequency of the quadrupole to 300 kHz and using it as a narrow-band mass filter, we show that it is possible to isolate ions from a single peak at m/z 22 000 in a window as narrow as 22 m/z units. Using cesium iodide cluster signals, we show that the mass range in the time-of-flight (TOF) analyzer extends beyond m/z 90 000, in theory to more than m/z 150 000. We also demonstrate that the resolution of the instrument is greater than 3000 at m/z 44 500. Tandem-MS capabilities are illustrated by separating components from heterooligomeric assemblies formed between tetrameric transthyretin, thyroxine, retinol-binding protein, and retinol. Isolation of a single charge state at m/z 5340 in the quadrupole and subsequent collision-induced dissociation (CID) in the gas-filled collision cell leads to the formation of ions from individual subunits and subcomplexes, identified by their mass and charge in the TOF analyzer.