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Related Experiment Video

Updated: Jul 11, 2026

A Quantitative Glycomics and Proteomics Combined Purification Strategy
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A Quantitative Glycomics and Proteomics Combined Purification Strategy

Published on: March 8, 2016

Non-gel-based dual 18O labeling quantitative proteomics strategy.

Huiling Liu1, Yangjun Zhang, Lingyan Meng

  • 1State Key Lab of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, 33 Zhongguancun Life Park Road, Beijing 102206, China.

Analytical Chemistry
|September 18, 2007
PubMed
Summary

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A novel dual (18)O labeling strategy using DTPA dianhydride enhances proteomic quantitation. This non-gel-based method improves accuracy and confidence by minimizing isotope exchange and MS interference.

Area of Science:

  • Proteomics
  • Analytical Chemistry
  • Biochemistry

Background:

  • Accurate protein quantitation is crucial for proteomic analysis.
  • Existing (18)O labeling methods have limitations, including back-exchange and isotope peaks.
  • Non-gel-based strategies are desirable for streamlined workflows.

Purpose of the Study:

  • To develop a novel non-gel-based, dual (18)O labeling strategy for improved protein quantitation.
  • To utilize diethylenetriamine-N,N,N', N' ',N' '-pentaacetic acid (DTPA) dianhydride for chemical (18)O labeling.
  • To combine chemical and enzyme-catalyzed (18)O labeling for enhanced accuracy.

Main Methods:

  • A dual (18)O labeling strategy employing DTPA dianhydride was developed.
  • Method 1: Chemical (18)O labeling via peptide amine acylation and hydrolysis.

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Last Updated: Jul 11, 2026

A Quantitative Glycomics and Proteomics Combined Purification Strategy
11:38

A Quantitative Glycomics and Proteomics Combined Purification Strategy

Published on: March 8, 2016

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
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Published on: November 15, 2017

Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level
08:29

Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level

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  • Method 2: Combined chemical and enzyme-catalyzed (18)O labeling of peptide carboxyl-termini.

    • Main Results:

    • DTPA acylation was rapid, specific, and generated y-series ions in MS/MS.
    • Both (18)O labeling methods demonstrated accuracy within 0.1-10 (16)O/(18)O ratios and <20% deviation.
    • The first method prevented (18)O to (16)O back-exchange, and the second avoided MS isotope peaks.

    Conclusions:

    • The developed dual (18)O labeling strategy significantly improves peptide quantitation accuracy.
    • The non-gel-based approach offers advantages over current proteolytic methods.
    • Combining chemical and proteolytic (18)O labeling enhances confidence in quantitative proteomic results.