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Tandem Mass Spectrometry01:21

Tandem Mass Spectrometry

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Tandem mass spectrometry is a technique that uses multiple mass analyzers in series to obtain a higher selectivity and signal-to-noise ratio for the analyte. Instruments with multiple analyzers separated by an interaction cell enable secondary fragmentation and selected study of the fragment ions.
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The quadrupole mass analyzer consists of four cylindrical metal rods arranged in a diamond carrying a DC voltage and a radio-frequency AC voltage. The motion of ions through the quadrupole depends on the field strength, causing only ions of a certain m/z to resonate successfully and strike the detector at a given field strength. Though the transmission rate for these analyzers is high, the exact elemental composition of the sample is not determined because of low resolution; however, they are...
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Measurement of Ultrafast Vibrational Coherences in Polyatomic Radical Cations with Strong-Field Adiabatic Ionization
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Accelerating covalent binding studies: Direct mass shift measurement with acoustic ejection and TOF-MS.

Markus Stoeckli1, Han Wang2, Dieter Staab1

  • 1Novartis Pharma AG, Biomedical Research, Novartis Campus, Switzerland.

SLAS Technology
|October 25, 2024
PubMed
Summary

This study introduces a faster method for tracking chemical reactions using acoustic ejection and mass spectrometry, enabling real-time analysis of compound-protein binding without lengthy sample preparation. This innovation significantly boosts assay throughput for drug discovery.

Keywords:
Biomedical researchCovalent bindingElectrospray, HTSMass spectrometryNative MSScreening

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Area of Science:

  • Analytical Chemistry
  • Biochemistry
  • Mass Spectrometry

Background:

  • Traditional methods for tracking chemical reactions, like liquid chromatography-mass spectrometry (LC-MS), require extensive sample pre-treatment (desalting) due to incompatible buffers.
  • This pre-treatment step is time-consuming, with analysis times ranging from seconds to minutes per sample, limiting overall assay throughput and broad applicability.
  • Direct mass spectrometry (MS) injection is desirable for faster analysis but is hindered by buffer incompatibilities in existing assays.

Purpose of the Study:

  • To develop and validate a streamlined workflow for real-time measurement of covalent binding kinetics.
  • To overcome the throughput limitations of traditional LC-MS methods for chemical reaction monitoring.
  • To demonstrate the application of a novel acoustic ejection (AE) interface coupled with a time-of-flight mass spectrometer (MS) for direct sample analysis.

Main Methods:

  • Integration of an acoustic ejection (AE) interface with a time-of-flight mass spectrometer (MS) for direct sample introduction.
  • Development of a complete workflow encompassing assay setup, sample analysis, and data evaluation for covalent binding studies.
  • Utilized the SCIEX Echo® MS+ system with the ZenoTOF 7600 system for kinetic analysis of covalent binding.

Main Results:

  • The AE-MS system enables direct sample injection at rates up to one sample per second, drastically reducing analysis time compared to traditional methods.
  • Successfully established a complete workflow for real-time measurement of covalent binding of compounds to proteins.
  • This represents the first reported use of the SCIEX Echo® MS+ system with the ZenoTOF 7600 system for studying covalent binding kinetics.

Conclusions:

  • Combining acoustic ejection with time-of-flight mass spectrometry offers a direct, high-throughput alternative for monitoring chemical reactions, specifically covalent binding.
  • The developed workflow eliminates the need for sample pre-treatment, significantly accelerating analysis and enabling real-time kinetic studies.
  • This technological advancement holds promise for broader application in drug discovery and chemical biology research due to its speed and efficiency.