Structural characterization of heparins from different commercial sources
- 20 September 2011
- journal article
- research article
- Published by Springer Nature in Analytical and Bioanalytical Chemistry
- Vol. 401 (9), 2793-2803
- https://doi.org/10.1007/s00216-011-5367-7
Abstract
Seven commercial heparin active pharmaceutical ingredients and one commercial low molecular weight from different manufacturers were characterized with a view profiling their physicochemical properties. All heparins had similar molecular weight properties as determined by polyacrylamide gel electrophoresis (M N, 10–11 kDa; M W, 13–14 kDa; polydispersity (PD), 1.3–1.4) and by size exclusion chromatography (M N, 14–16 kDa; M W, 21–25 kDa; PD, 1.4–1.6). one-dimensional 1H- and 13C-nuclear magnetic resonance (NMR) evaluation of the heparin samples was performed, and peaks were fully assigned using two-dimensional NMR. The percentage of glucosamine residues with 3-O-sulfo groups and the percentage of N-sulfo groups and N-acetyl groups ranged from 5.8–7.9%, 78–82%, to 13–14%, respectively. There was substantial variability observed in the disaccharide composition, as determined by high performance liquid chromatography (HPLC)-mass spectral analysis of heparin lyase I–III digested heparins. Heparin oligosaccharide mapping was performed using HPLC following separate treatments with heparin lyase I, II, and III. These maps were useful in qualitatively and quantitatively identifying structural differences between these heparins. The binding affinities of these heparins to antithrombin III and thrombin were evaluated by using a surface plasmon resonance competitive binding assay. This study provides the physicochemical and activity characterization necessary for the appropriate design and synthesis of a generic bioengineered heparin.Keywords
This publication has 28 references indexed in Scilit:
- Impact of Autoclave Sterilization on the Activity and Structure of Formulated HeparinJournal of Pharmaceutical Sciences, 2011
- Ultra-performance ion-pairing liquid chromatography with on-line electrospray ion trap mass spectrometry for heparin disaccharide analysisAnalytical Biochemistry, 2011
- Control of the heparosan N-deacetylation leads to an improved bioengineered heparinApplied Microbiology and Biotechnology, 2011
- Mass balance analysis of contaminated heparin productAnalytical Biochemistry, 2011
- Heparin Mapping Using Heparin Lyases and the Generation of a Novel Low Molecular Weight HeparinJournal of Medicinal Chemistry, 2010
- E. coli K5 fermentation and the preparation of heparosan, a bioengineered heparin precursorBiotechnology & Bioengineering, 2010
- Orthogonal analytical approaches to detect potential contaminants in heparinProceedings of the National Academy of Sciences, 2009
- Solution Structures of Chemoenzymatically Synthesized Heparin and Its PrecursorsJournal of the American Chemical Society, 2008
- Oversulfated chondroitin sulfate is a contaminant in heparin associated with adverse clinical eventsNature Biotechnology, 2008
- Engineering sulfotransferases to modify heparan sulfateNature Chemical Biology, 2008