A Tandem Mass Spectrometer for Improved Transmission and Analysis of Large Macromolecular Assemblies
Abstract
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 3,000-4,000, 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 3,000 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 5,340 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.
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Native MS instruments can transmit and detect high-m/z ions using commercially available time-of-flight, Orbitrap, and Fourier transform ion cyclotron resonance instruments.
“Today, this is possible using commercially available time-of-flight,33 Orbitrap,34 and Fourier transform ion cyclotron resonance (FTICR)35 instruments.”
From Higher-order structural characterisation of native proteins and complexes by top-down mass spectrometry (2020) · cites this paper as [33]
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- Authors
- Sobott, F., Hernández, H., McCammon, M. G., Tito, M. A., & Robinson, C. V.
- Affiliations
- University of Cambridge (GB)
- Venue
- Analytical Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2002-02-16
- Volume
- 74
- Issue
- 6
- Pages
- 1402-1407
- ISSN
- 0003-2700 (ISSN-L)
- 1520-6882
- OpenAlex
- W2066011752
- Cited by
- 511
- References
- 19
- Citations by year
- 2026: 4 · 2025: 11 · 2024: 11 · 2023: 18 · 2022: 11 · 2021: 26 · 2020: 21 · 2019: 20 · 2018: 18 · 2017: 27 · 2016: 25 · 2015: 29 · 2014: 34 · 2013: 26 · 2012: 24
- FWCI
- 8.67
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Analytical Chemistry (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Royal Society
- Medical Research Council
- Engineering and Physical Sciences Research Council
- Biotechnology and Biological Sciences Research Council
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolomics and Mass Spectrometry Studies
- Keywords
- tandem mass spectrometry, ion transmission, quadrupole frequency, prealbumin, thyroxine, retinol-binding protein, retinol, collision-induced dissociation
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