Recently, certain lots of heparin have been associated with an acute, rapid onset of serious side effects indicative of an allergic-type reaction. To identify potential causes for this sudden rise in side effects, we examined lots of heparin that correlated with adverse events using orthogonal high-resolution analytical techniques. Through detailed structural analysis, the contaminant was found to contain a disaccharide repeat unit of glucuronic acid linked β1→3 to a β-N-acetylgalactosamine. The disaccharide unit has an unusual sulfation pattern and is sulfated at the 2-O and 3-O positions of the glucuronic acid as well as at the 4-O and 6-O positions of the galactosamine. Given the nature of this contaminant, traditional screening tests cannot differentiate between affected and unaffected lots. Our analysis suggests effective screening methods that can be used to determine whether or not heparin lots contain the contaminant reported here.
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The authors would like to thank Sucharita Roy for work on the chemical synthesis of oversulfated chondroitin sulfate standards, Scott Bailey for assistance in the analysis of selected samples by one- and two-dimensional NMR and Laura Citterio for work in various separation and analytical steps. We also thank James Anderson, Josh Sorafine and Andre Jones for design of composition experiments and HPLC analysis of the heparinase-digested samples. Finally, we thank Ada Ziolkowski for help with manuscript preparation. This work was supported in part by US National Institute of General Medical Sciences grants GM57073 (R.S.) and GM38060 (R.J.L).
R.L. and R.S. have been directors of Momenta Pharmaceuticals since 2001 and own stock in the company, and both of them have served as scientific advisors to Scientific Protein Labs (SPL) since the end of February 2008. R.J.L. has served as a scientific advisor to Baxter International since mid-February 2008.
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Guerrini, M., Beccati, D., Shriver, Z. et al. Oversulfated chondroitin sulfate is a contaminant in heparin associated with adverse clinical events. Nat Biotechnol 26, 669–675 (2008). https://doi.org/10.1038/nbt1407
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