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Tandem mass spectrometry is a technique that uses multiple mass analyzers in series to obtain a higher selectivity and reduce chemical noise during analyte detection. Instruments with multiple analyzers separated by an interaction cell enable secondary fragmentation and selected study of the fragment ions.Secondary fragmentations occur in the interaction cell and can be induced by various factors. Fragmentation induced by collision with inert gases, such as N2, Ar, He, etc., is called...
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Analyzing Large Protein Complexes by Structural Mass Spectrometry
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Expanding the structural analysis capabilities on an Orbitrap-based mass spectrometer for large macromolecular

Kyle L Fort1, Michiel van de Waterbeemd, Dmitriy Boll

  • 1Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute of Pharmaceutical Sciences, Utrecht University, 3584 Utrecht, The Netherlands. a.j.r.heck@uu.nl.

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Native mass spectrometry is crucial for analyzing large biological molecules. This study details modifications to an Orbitrap mass spectrometer to improve its performance for complex analyses.

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Area of Science:

  • Biophysical Chemistry
  • Analytical Chemistry
  • Structural Biology

Background:

  • Native mass spectrometry (MS) is a powerful technique for characterizing macromolecular biological systems.
  • Increasing complexity and size of analytes necessitate advancements in instrument capabilities for accurate structural determination.

Purpose of the Study:

  • To describe modifications to an Orbitrap Q Exactive Plus mass spectrometer.
  • To enhance instrument performance for analyzing large and complex biomolecules.

Main Methods:

  • Detailed description of instrumental modifications to the Orbitrap Q Exactive Plus.
  • Implementation of advanced data acquisition and processing techniques.

Main Results:

  • Demonstrated increases in signal intensity for large analytes.
  • Improved mass resolution enabling finer structural details.
  • Expanded maximum m/z (mass-to-charge ratio) range for broader applicability.

Conclusions:

  • The described modifications significantly enhance the capabilities of the Orbitrap Q Exactive Plus for native MS.
  • These advancements facilitate more comprehensive structural analysis of large and complex biological systems.