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Related Experiment Video

Updated: Apr 20, 2026

Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry
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Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry

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Protein quantification using a cleavable reporter peptide.

Elodie Duriez1, Stephane Trevisiol, Bruno Domon

  • 1Luxembourg Clinical Proteomics Center, CRP-Santé , 1 A-B rue Thomas Edison, Strassen 1445, Luxembourg.

Journal of Proteome Research
|November 21, 2014
PubMed
Summary
This summary is machine-generated.

A new method rapidly recalibrates stable isotope labeled peptides for accurate protein quantification. This approach uses concatenated polypeptide standards for reliable biomarker measurement in complex samples.

Keywords:
calibrationcleavable reporter peptidemass spectrometrypeptide/protein quantificationstable isotope labeled peptidesstandard recalibrationtargeted proteomics

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Mass Spectrometry

Background:

  • Peptide and protein quantification using isotope dilution and mass spectrometry is crucial for biomarker measurement and systems biology.
  • The accuracy of these quantitative assays relies heavily on the quality of stable-isotope labeled standards, including purity, stability, and concentration determination.
  • Existing methods can be limited by biases in internal standard quality.

Purpose of the Study:

  • To develop a rapid, cost-efficient method for recalibrating stable isotope labeled peptides.
  • To improve the accuracy and reliability of protein quantification assays.
  • To introduce a novel concatenated polypeptide standard for enhanced quantitative analysis.

Main Methods:

  • A single liquid chromatography-mass spectrometry (LC-MS) analysis was developed for recalibration.
  • The method utilizes the equimolar release of a protein reference peptide and a universal reporter peptide from a concatenated polypeptide standard during trypsinization.
  • This approach serves as a surrogate for the protein of interest and allows for simultaneous recalibration.

Main Results:

  • The study demonstrates a rapid and cost-efficient recalibration of stable isotope labeled peptides.
  • Concatenated polypeptide standards were shown to generate high-quality and accurate quantitative data.
  • The method was successfully applied to quantify clinically important proteins in urine samples, showing comparable results to conventional methods.

Conclusions:

  • The developed method offers a significant improvement for the accurate quantification of peptides and proteins.
  • Concatenated polypeptide standards provide a reliable and efficient tool for recalibrating stable isotope labeled peptides.
  • This approach enhances the application of quantitative mass spectrometry in clinical and systems biology research.