Crude Heparin Preparations Unveil the Presence of Structurally Diverse Oversulfated Contaminants

Abstract

Nowadays, pharmaceutical heparin is purified from porcine and bovine intestinal mucosa. In the past decade there has been an ongoing concern about the safety of heparin, since in 2008, adverse effects associated with the presence of an oversulfated chondroitin sulfate (OSCS) were observed in preparations of pharmaceutical porcine heparin, which led to the death of patients, causing a global public health crisis. However, it has not been clarified whether OSCS has been added to the purified heparin preparation, or whether it has already been introduced during the production of the raw heparin. Using a combination of different analytical methods, we investigate both crude and final heparin products and we are able to demonstrate that the sulfated contaminants are intentionally introduced in the initial steps of heparin preparation. Furthermore, the results show that the oversulfated compounds are not structurally homogeneous. In addition, we show that these contaminants are able to bind to cells in using well known heparin binding sites. Together, the data highlights the importance of heparin quality control even at the initial stages of its production.

Anticoagulation Strategies During ECMO Application

“…In addition to the risk of HIT and the shortcomings of heparin as an anticoagulant, the contamination crisis during heparin use in 2008 has led to investigations into nonanimal-derived heparin-like anticoagulant sources [19].…”

Pharmacology of Heparin and Related Drugs: An Update

“…Since the episode of contamination of pharmaceutical heparin with OSCS in 2007-2008 that led to serious adverse events associated with the clinical use of certain heparin preparations, including fatalities (Chess et al, 2012), development of new methods for the assessment of heparin continues (Devlin et al, 2019). It has now been established that the contaminant OSCS was added at an early stage of heparin manufacture, so methods applicable to the efficient screening of crude heparin samples rather than API and final product are particularly useful (Mauri et al, 2017b;Mendes et al, 2019) in ensuring such contamination does not occur in the future.…”

Cell-surface glycosaminoglycans regulate the cellular uptake of charged polystyrene nanoparticles

“…Chondroitinase ABC was obtained from Seikagaku Corporation (Tokyo, Japan), whereas Heparitinase II was pre-pared from Flavobacterium heparinum as reported elsewhere. 80,81 Chlorpromazine hydrochloride, 5-(N-ethyl-Nisopropyl)amiloride, PP2, nystatin, filipin, dynasore, polyinosinic acid and polyadenylic acid were from Sigma-Aldrich, while Alexa Fluor 488-labeled transferrin, FITC-labeled 70 kDa dextran, BODIPY FL C5-lactosylceramide/BSA complex (LacCer), and Alexa Fluor 488-labeled wheat germ agglutinin were from Thermo Fisher Scientific. All remaining reagents were from Sigma-Aldrich.…”