Porcine and Ovine Mucosal Heparins and Their Depolymerized Derivatives Are Comparable in Contrast to Their Bovine Equivalents
Authors: Bouchard Olivia , Lewis Mary , Hoppensteadt Debra , Maia Paula , de Castro Alice Silva , Kumar Emmanuele , Guler Nil , Jeske Walter , Kahn Daniel , Walenga Jeanine , Coyne Erwin , Fareed Jaweed 
Published: November 2023

Introduction

The currently used unfractionated heparin (UFH) and low molecular weight heparins (LMWH) are mostly derived from Porcine mucosal tissue. Since the technology to manufacture heparin has advanced and quality assurance practices are in place, improved products with high potency and purity are now available. The purpose of this study is to compare multiple individual batches of UFH obtained from Bovine, Ovine, and Porcine origin and their depolymerized product obtained by benzylation followed by alkaline hydrolysis representing enoxaparins.

Methods

The molecular profile of the heparins and enoxaparins from various sources were determined using the size exclusion method. The anticoagulant potency was measured using clot based methods such as aPTT and Thrombin Time. Chromogenic substrate based methods were used to determine the USP potency in terms of anti‐Xa and anti‐IIa activities (Hyphen Biomedical, Ohio, USA). The relative interaction of the heparins and enoxaparins with heparin induced thrombocytopenia (HIT) antibody induced aggregation of platelets were investigated using aggregometry.

Results

The molecular profiles of the Bovine, ovine, and porcine heparins and enoxaparins were almost comparable and ranged from 15–18 kDa. The global anticoagulant and amidolytic protease assays for the bovine heparin were consistently lower than porcine and ovine samples. In the purified system the Porcine and Ovine preparations consistently showed lower IC50 values for both the thrombin and Xa inhibition in contrast to the bovine heparin. Similar trends were observed in the anti IIa assays. The USP potency of 28 batches of porcine ranged from 170–200 U/mg, whereas the 21 batches of ovine heparins exhibited comparable potencies in the range of 160–210 U/mL. The USP potency of bovine mucosal heparin was much lower and ranged from 110–140 U/mg. The anti‐Xa – IIa ratio for the porcine and ovine heparins were comparable. However, the anti‐Xa – IIa ratio of bovine heparin was somewhat lower. The ovine and porcine enoxaparin exhibited comparable potencies which ranged from 94–110 U/mg whereas bovine enoxaparin was slightly lower, ranging from 80–87 U/mg. However the antiXa and anti‐IIa ratios of the enoxaparins derived from various species were comparable. In the HIT screening, there was no difference between the HIT responses in the heparins from different species. Similar results were obtained amongst enoxaparins of different origins.

Conclusions

These studies show that heparins from bovine, ovine, and porcine origin exhibit comparable molecular profiles. While the porcine and ovine heparins exhibit similar biological potencies, the bovine heparin was found to be weaker. The enoxaparins derived from these species also exhibit similar molecular profiles, however, in functional assays, they showed weaker potency. In the HIT screening profile, the heparins from all of these species produced stronger effects in comparison to their enoxaparin counterparts.