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Related Concept Videos

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.
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 collision-induced...
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Mass Spectrometry: Complex Analysis01:21

Mass Spectrometry: Complex Analysis

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Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
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Mass Analyzers: Common Types01:19

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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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Mass Spectrometers01:16

Mass Spectrometers

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This lesson details the instrumentation of a mass spectrometer—a physical instrument to perform mass spectrometry on analyte molecules and record the characteristic mass spectra. This is achieved via three chief functions:
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Peptide Identification Using Tandem Mass Spectrometry01:33

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Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
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Updated: Jun 13, 2025

Label-Free Immunoprecipitation Mass Spectrometry Workflow for Large-scale Nuclear Interactome Profiling
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Standardized workflow for multiplexed charge detection mass spectrometry on orbitrap analyzers.

Pei Su1, John P McGee1,2,3, Michael A R Hollas3

  • 1Departments of Molecular Biosciences, Chemistry and Chemical and Biological Engineering and the Feinberg School of Medicine, Northwestern University, Evanston, IL, USA.

Nature Protocols
|January 3, 2025
PubMed
Summary

Individual ion mass spectrometry (I²MS) extends charge detection mass spectrometry to Orbitrap instruments. This method directly measures ion mass and charge, enabling mass spectral output without deconvolution for complex samples.

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Area of Science:

  • Analytical Chemistry
  • Biochemistry
  • Mass Spectrometry

Background:

  • Charge detection mass spectrometry (CDMS) is a specialized technique for analyzing individual ions.
  • Traditional CDMS is limited to custom-built instruments.
  • Orbitrap mass analyzers offer advanced capabilities for mass spectrometry.

Purpose of the Study:

  • To extend CDMS capabilities to Orbitrap analyzers, termed Individual ion mass spectrometry (I²MS).
  • To develop a standardized workflow for I²MS data acquisition and analysis.
  • To enable direct mass spectral output without deconvolution for complex biological samples.

Main Methods:

  • Protein sample preparation and signal attenuation for individual ion detection.
  • Charge calibration using standard proteins with known charge states.
  • Data analysis using STORIboard software for mass spectral output generation.

Main Results:

  • Simultaneous measurement of mass-to-charge ratio (m/z) and charge (z) for individual ions.
  • Direct generation of mass distribution profiles for heterogeneous samples.
  • Successful analysis of complex mixtures, including intact monoclonal antibodies and their glycosylation patterns.

Conclusions:

  • I²MS provides a powerful extension of CDMS for Orbitrap users.
  • The developed workflow facilitates the analysis of complex samples intractable by traditional methods.
  • I²MS enables detailed characterization of biomolecules like monoclonal antibodies and their modifications.