Impact of autoclave sterilization on the activity and structure of formulated heparin

Loading...
Thumbnail Image
Authors
Beaudet, Julie M.
Weyers, Amanda
Solakyildirim, Kemal
Yang, Bo
Takieddin, Majde
Mousa, Shaker
Zhang, Fuming
Linhardt, Robert J.
Issue Date
2011-01-01
Type
Article
Language
ENG
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
Research Projects
Organizational Units
Journal Issue
Alternative Title
Abstract
The stability of a formulated heparin was examined during its sterilization by autoclaving. A new method to follow loss in heparin binding to the serine protease inhibitor, antithrombin III, and the serine protease, thrombin, was developed using a surface plasmon resonance competitive binding assay. This loss in binding affinity correlated well with loss in antifactor IIa (thrombin) activity as well as antifactor Xa activity as measured using conventional amidolytic assays. Autoclaving also resulted in a modest breakdown of the heparin backbone as confirmed by a slight reduction in number-averaged and weight-averaged molecular weight and an increase in polydispersity. Although no clear changes were observed by nuclear magnetic resonance spectroscopy, disaccharide composition analysis using high-performance liquid chromatography-electrospray ionization-mass spectrometry suggested that loss of selected sulfo groups had taken place. It is this sulfo group loss that probably accounts for a decrease in the binding of autoclaved heparin to antithrombin III and thrombin as well as the observed decrease in its amidolytic activity.
Description
Journal of Pharmaceutical Science, 100, 3396–3404
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
Full Citation
Impact of autoclave sterilization on the activity and structure of formulated heparin, J. M. Beaudet, A. Weyers, K. Solakyildirim, B. Yang, M. Takieddin, S. Mousa, F. Zhang, R. J. Linhardt, Journal of Pharmaceutical Science, 100, 3396–3404, 2011.
Publisher
Elsevier
Terms of Use
Journal
Volume
Issue
PubMed ID
DOI
ISSN
15206017
223549
EISSN