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Improving the Sensitivity for Quantifying Heparan Sulfate from Biological Samples.

Zhangjie Wang1, Vijay M Dhurandhare1,2, Cressida A Mahung3

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Summary

A new method using AMQC labeling allows sensitive quantification of heparan sulfates (HSs) in biological samples. This technique aids in disease prognosis by profiling HS levels in animal models of liver and burn injuries.

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Area of Science:

  • Glycoscience
  • Analytical Chemistry
  • Biomarker Discovery

Background:

  • Heparan sulfates (HSs) are crucial glycans involved in numerous biological processes, including cell regulation and disease.
  • Accurate quantification and structural analysis of HS are vital for understanding its in vivo functions.
  • Current methods for HS analysis lack the sensitivity and reliability needed for comprehensive investigation.

Purpose of the Study:

  • To develop a novel, sensitive, and reliable quantitative technique for analyzing heparan sulfate (HS) disaccharides.
  • To introduce a new labeling reagent, 6-amino-N-(2-diethylamino)ethyl quinoline-2-carboamide (AMQC), for HS analysis.
  • To apply the developed method for HS profiling in animal models to assess disease prognosis.

Main Methods:

  • Development of a new labeling reagent, AMQC, for heparan sulfate disaccharides.
  • Analysis of AMQC-conjugated disaccharides using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in positive mode.
  • Utilizing 13C-labeled HS disaccharide internal standards for accurate quantification and composition determination.

Main Results:

  • The AMQC labeling significantly enhanced the sensitivity of HS disaccharide analysis by LC-MS/MS.
  • The method enabled precise determination of HS amount and disaccharide composition from single histological slides.
  • HS levels were successfully profiled in plasma/serum and tissues/organs of mouse models for liver and burn injuries.

Conclusions:

  • The developed AMQC-based LC-MS/MS method provides a sensitive and reliable tool for HS analysis.
  • This technique can profile HS levels in various biological samples, aiding in disease prognosis.
  • The findings suggest HS may play a role in disease pathogenesis and could serve as potential diagnostic biomarkers.