Structural characterization of heparins from different commercial sources.
Anal Bioanal Chem
; 401(9): 2793-803, 2011 Nov.
Article
in En
| MEDLINE
| ID: mdl-21931955
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 (1)H- and (13)C-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.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Heparin
/
Magnetic Resonance Spectroscopy
/
Chemistry, Physical
/
Chromatography, High Pressure Liquid
/
Surface Plasmon Resonance
Type of study:
Prognostic_studies
Limits:
Animals
Language:
En
Journal:
Anal Bioanal Chem
Year:
2011
Document type:
Article
Affiliation country:
Estados Unidos