Competitive inhibition of heparinase by persulfonated glycosaminoglycans: a tool to detect heparin contamination
Research article published in Analytical chemistry (2011)
Abstract
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.
Summary
Heparin and the low molecular weight heparins are extensively used as medicinal products to prevent and treat the formation of venous and arterial thrombi.
Why This Matters for Hirudotherapy
This study examined whether enzymatic depolymerization using bacterially derived heparinases could detect oversulfated chondroitin sulfate (OSCS) contamination in heparin products, following the 2008 adverse events associated with contaminated heparin lots. The investigators found that persulfonated glycosaminoglycans were effective inhibitors of heparinase I (IC50 values approximately 0.5–2 μg/mL) and developed a heparinase-digestion assay with size-exclusion HPLC that could routinely detect less than 0.1% (w/w) OSCS in heparin. For ASH/hirudotherapy, this work underscores the critical importance of rigorous purity testing for parenteral anticoagulant products—a principle directly relevant to leech-derived therapeutics such as hirudin, where contamination could similarly cause serious adverse reactions. The caveat is that this study specifically addresses heparin quality control methodology and does not directly evaluate leech secretome components or their clinical applications.
Citation
Competitive inhibition of heparinase by persulfonated glycosaminoglycans: a tool to detect heparin contamination
Aich U et al. · Analytical chemistry, 2011
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