A simple method to measure dermatan sulfate at sub-microgram concentrations in plasma

Area of Science:

• Biochemistry
• Pharmacology
• Hematology

Background:

• Dermatan sulfate (DS) is a glycosaminoglycan with potential therapeutic applications.
• Accurate measurement of DS in biological fluids is crucial for research.
• Existing methods may lack sensitivity or specificity.

Purpose of the Study:

• To develop a simple and sensitive biological assay for quantifying dermatan sulfate (DS) in plasma.
• To establish the assay's reliability and applicability across different species.
• To assess the ex vivo anticoagulant effects of DS.

Main Methods:

• Assay principle: measuring residual thrombin activity after incubation with heparin cofactor II (HC II) in defibrinated plasma.
• Exploiting low ionic strength to enhance thrombin inhibition rate.
• Utilizing bentonite absorption to remove competing plasma inhibitors.

Main Results:

• A linear relationship was observed between DS concentrations (0-2 µg/ml) and the log of residual thrombin activity.
• The assay demonstrated a sensitivity limit of 0.1 µg/ml.
• The method showed acceptable reproducibility and sensitivity to other HC II activators like heparin.

Conclusions:

• A simple, sensitive, and reproducible biological assay for plasma dermatan sulfate (DS) has been developed.
• The assay is effective for human, rabbit, and rat plasma and aids in studying DS and related glycosaminoglycans.
• Dermatan sulfate exhibits efficacy in experimental thrombosis without significant ex vivo anticoagulant activity.