Heparin microarray-based specific interaction assay of marine sulfated polysaccharides with antithrombin Ⅲ

Area of Science:

• Biochemistry
• Pharmacology
• Materials Science

Background:

• Marine sulfated polysaccharides (SPs) are investigated as potential anticoagulant drugs.
• Efficient molecular-level methods are needed to assess their anticoagulant activity.
• Understanding SP interactions with antithrombin III (AT) is crucial for drug development.

Purpose of the Study:

• To develop a heparin microarray-based competitive strategy to evaluate specific interactions between SPs and AT.
• To determine the inhibitory concentrations (IC50) of various SPs against AT.
• To assess the versatility of the strategy for evaluating SP interactions with other proteins.

Main Methods:

• A competitive binding assay using heparin microarrays to measure AT displacement by SPs.
• Optimization of experimental conditions for the microarray assay.
• Validation using surface plasmon resonance (SPR) and established anticoagulants (heparin, enoxaparin).

Main Results:

• The developed method successfully determined IC50 values for three SPs: fucoidan (AnF), fucosylated chondroitin sulfate (FCSht), and its degraded form (S-dFCSht).
• AnF showed an IC50 of 50.55 ± 2.79 μg/mL, FCSht of 44.18 ± 4.05 μg/mL, and S-dFCSht of 16.99 ± 6.56 μg/mL.
• The strategy demonstrated versatility by assessing SP interactions with side effect-related proteins.

Conclusions:

• The heparin microarray-based competitive strategy provides an efficient method for evaluating anticoagulant activity of SPs at the molecular level.
• The method offers insights into both the efficacy and potential safety profiles of SPs.
• This strategy can be adapted for evaluating SP interactions with various anticoagulant targets by creating specialized glycan microarrays.