Competitive inhibition of heparinase by persulfonated glycosaminoglycans: a tool to detect heparin contamination

Heparin and the low molecular weight heparins are extensively used as medicinal products to prevent and treat the formation of venous and arterial thrombi. In early 2008, administration of some heparin lots was associated with the advent of severe adverse effects, indicative of an anaphylactoid-like response. Application of orthogonal analytical tools enabled detection and identification of the contaminant as oversulfated chondroitin sulfate (OSCS) was reported in our earlier report. Herein, we investigate whether enzymatic depolymerization using the bacterially derived heparinases, given the structural understanding of their substrate specificity, can be used to identify the presence of OSCS in heparin. We also extend this analysis to examine the effect of other persulfonated glycosaminoglycans (GAGs) on the action of the heparinases. We find that all persulfonated GAGs examined were effective inhibitors of heparinase I, with IC(50) values ranging from approximately 0.5-2 μg/mL. Finally, using this biochemical understanding, we develop a rapid, simple assay to assess the purity of heparin using heparinase digestion followed by size-exclusion HPLC analysis to identify and quantify digestion products. In the context of the assay, we demonstrate that less than 0.1% (w/w) of OSCS (and other persulfonated polysaccharides) can routinely be detected in heparin.

Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.

Heparin and the low molecular weight heparins are extensively used as medicinal products to prevent and treat the formation of venous and arterial thrombi.

Explore how this research connects to clinical practice

More peer-reviewed studies in Antimicrobial Resistance.

• Novel Leech Antimicrobial Peptides — Hirunipins — Kumar et al. (2025)
• AIF-1 and RNASET2 Play Complementary Roles in the Innate Immune Response of Medicinal Leech — Baranzini N et al. (2019)
• Aeromonas septicemia after medicinal leech use following replantation of severed digits — Levine SM et al. (2009)
• Low-Level Antimicrobials in the Medicinal Leech Select for Resistant Pathogens — Beka L et al. (2018)
• Role of Antimicrobial Peptides in Immunity of Parasitic Leeches — Kaygorodova IA (2023)
• The elastase-inhibitor eglin has no effect in an ovine model of endotoxemia — Redl H et al. (1988)
• A novel Toll like receptor with two TIR domains (HcToll-2) is involved in regulation of antimicrobial peptide gene expression of Hyriopsis cumingii. — Ren Q et al. (2014)
• Molecular cloning and functional characterization of a short peptidoglycan recognition protein from triangle-shell pearl mussel (Hyriopsis cumingii). — Huang Y et al. (2018)
• Isolation, Characterization, and Evaluation of a Lytic Jumbo Phage Z90 Againstin American Eels (). — Zhang M et al. (2025)
• Elucidating environmental reservoir of antimicrobial resistance - a phenotypic characterization of gut microbiota from aquatic coleoptera in a low-anthropogenic impact zone. — Orhan F et al. (2025)

Browse all ASH-analysed research →