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Related Concept Videos

Mass Analyzers: Common Types01:19

Mass Analyzers: Common Types

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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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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.
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Updated: Jun 18, 2025

An HS-MRM Assay for the Quantification of Host-cell Proteins in Protein Biopharmaceuticals by Liquid Chromatography Ion Mobility QTOF Mass Spectrometry
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Rapid assay development for low input targeted proteomics using a versatile linear ion trap.

Brian Searle1, Ariana Shannon1, Rachael Teodorescu1

  • 1Ohio State University.

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Summary
This summary is machine-generated.

This study introduces a cost-effective hybrid quadrupole-linear ion trap (LIT) mass spectrometry workflow for targeted proteomics. It enables rapid assay development from global data-independent acquisition, democratizing advanced proteomic analysis for limited cell populations.

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Area of Science:

  • Proteomics
  • Mass Spectrometry
  • Cellular Biology

Background:

  • High-mass accuracy instruments are typically required for studying limited cell populations in proteomics.
  • Triple quadrupoles are limited to targeted proteomics due to low-mass accuracy measurements.
  • Linear ion traps (LITs) provide a versatile, cost-effective option for both targeted and global proteomics.

Purpose of the Study:

  • To describe a workflow for developing targeted proteomics assays from global data-independent acquisition (DIA) measurements using a hybrid quadrupole-LIT instrument.
  • To demonstrate the capability of this workflow without requiring high-mass accuracy.
  • To showcase the instrument's utility for analyzing limited cell populations and low-level proteins.

Main Methods:

  • Utilized a new hybrid quadrupole-linear ion trap (LIT) instrument.
  • Developed an automated software approach for scheduling parallel reaction monitoring (PRM) assays.
  • Performed global data-independent acquisition (DIA) for assay development.
  • Quantified low-level proteins in a 1 ng background proteome.

Main Results:

  • Achieved consistent quantification across three orders of magnitude in a matched-matrix background.
  • Demonstrated quantitative linearity below two orders of magnitude for low-level proteins like transcription factors and cytokines, without stable isotope-labeled standards.
  • Showed consistency between proteins in CD4+ and CD8+ T cell subsets using LIT-based proteomics and high-dimensional flow cytometry from a 1 ng sample.

Conclusions:

  • Hybrid quadrupole-LIT instruments offer an economical solution for targeted proteomics.
  • This workflow democratizes mass spectrometry for diverse laboratory settings, especially for limited sample sizes.
  • The method enables sensitive quantification of low-abundance proteins crucial for cellular studies.