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Updated: Jun 6, 2026

Protease- and Acid-catalyzed Labeling Workflows Employing 18O-enriched Water
09:43

Protease- and Acid-catalyzed Labeling Workflows Employing 18O-enriched Water

Published on: February 20, 2013

Accelerated 18O-labeling in urinary proteomics.

Håvard Loftheim1, Anders Asberg, Léon Reubsaet

  • 1Department of Pharmaceutical Chemistry, School of Pharmacy, University of Oslo, P.O. Box 1068 Blindern, NO-0316 Oslo, Norway.

Journal of Chromatography. A
|November 25, 2010
PubMed
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This study optimized oxygen-18 (18O) labeling for faster peptide analysis in proteomics. An integrated method using immobilized trypsin achieved efficient labeling within 3.5 hours, improving sample throughput for biomarker discovery.

Area of Science:

  • Proteomics
  • Analytical Chemistry
  • Biochemistry

Background:

  • Proteolytic (18)O-labeling is crucial for quantitative proteomics.
  • Optimizing labeling efficiency and speed is essential for high-throughput analysis.
  • Current methods can be time-consuming and involve multiple sample transfer steps.

Purpose of the Study:

  • To enhance the efficiency and speed of proteolytic (18)O-labeling.
  • To develop an integrated approach for tryptic digestion and (18)O/(16)O-labeling.
  • To validate the method in complex biological matrices for biomarker discovery.

Main Methods:

  • Utilized tryptic digestion of bovine serum albumin (BSA) and cytochrome c peptides.
  • Employed immobilized trypsin for both in-solution digestion and subsequent (18)O/(16)O-labeling.

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Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
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Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

Published on: November 15, 2017

Related Experiment Videos

Last Updated: Jun 6, 2026

Protease- and Acid-catalyzed Labeling Workflows Employing 18O-enriched Water
09:43

Protease- and Acid-catalyzed Labeling Workflows Employing 18O-enriched Water

Published on: February 20, 2013

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
10:37

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

Published on: November 15, 2017

  • Integrated digestion (pH 8) and labeling (pH 6) steps without sample transfer.

    • Main Results:

    • Achieved complete (18)O-labeling within 2 hours at pH 6 in initial experiments.
    • Developed an integrated approach completing digestion and labeling in 3.5 hours with high (18)O/(16)O-ratios (0.96±0.13).
    • Validated the method in spiked urine samples and a full-scale bottom-up experiment, showing good performance (ratios 0.83±0.13 and 0.91±0.27).

    Conclusions:

    • The integrated approach using immobilized trypsin significantly simplifies sample handling and reduces reaction time.
    • This optimized method demonstrates high efficiency and accuracy for (18)O/(16)O-labeling in proteomics.
    • The approach is suitable for complex matrices like urine, supporting biomarker discovery applications.