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Heparanase as an Additional Tool for Detecting Structural Peculiarities of Heparin Oligosaccharides

Due to the biological properties of heparin and low-molecular-weight heparin (LMWH), continuous advances in elucidation of their microheterogeneous structure and discovery of novel structural peculiarities are crucial. Effective strategies for monitoring manufacturing processes and assessment of mor...

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Autores principales: Alekseeva, Anna, Urso, Elena, Mazzini, Giulia, Naggi, Annamaria
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6930493/
https://www.ncbi.nlm.nih.gov/pubmed/31810297
http://dx.doi.org/10.3390/molecules24234403
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author Alekseeva, Anna
Urso, Elena
Mazzini, Giulia
Naggi, Annamaria
author_facet Alekseeva, Anna
Urso, Elena
Mazzini, Giulia
Naggi, Annamaria
author_sort Alekseeva, Anna
collection PubMed
description Due to the biological properties of heparin and low-molecular-weight heparin (LMWH), continuous advances in elucidation of their microheterogeneous structure and discovery of novel structural peculiarities are crucial. Effective strategies for monitoring manufacturing processes and assessment of more restrictive specifications, as imposed by the current regulatory agencies, need to be developed. Hereby, we apply an efficient heparanase-based strategy to assert the structure of two major isomeric octasaccharides of dalteparin and investigate the tetrasaccharides arising from antithrombin binding region (ATBR) of bovine mucosal heparin. Heparanase, especially when combined with other sample preparation methods (e.g., size exclusion, affinity chromatography, heparinase depolymerization), was shown to be a powerful tool providing relevant information about heparin structural peculiarities. The applied approach provided direct evidence that oligomers bearing glucuronic acid–glucosamine-3-O-sulfate at their nonreducing end represent an important structural signature of dalteparin. When extended to ATBR-related tetramers of bovine heparin, the heparanase-based approach allowed for elucidation of the structure of minor sequences that have not been reported yet. The obtained results are of high importance in the view of the growing interest of regulatory agencies and manufacturers in the development of low-molecular-weight heparin generics as well as bovine heparin as alternative source.
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spelling pubmed-69304932019-12-26 Heparanase as an Additional Tool for Detecting Structural Peculiarities of Heparin Oligosaccharides Alekseeva, Anna Urso, Elena Mazzini, Giulia Naggi, Annamaria Molecules Article Due to the biological properties of heparin and low-molecular-weight heparin (LMWH), continuous advances in elucidation of their microheterogeneous structure and discovery of novel structural peculiarities are crucial. Effective strategies for monitoring manufacturing processes and assessment of more restrictive specifications, as imposed by the current regulatory agencies, need to be developed. Hereby, we apply an efficient heparanase-based strategy to assert the structure of two major isomeric octasaccharides of dalteparin and investigate the tetrasaccharides arising from antithrombin binding region (ATBR) of bovine mucosal heparin. Heparanase, especially when combined with other sample preparation methods (e.g., size exclusion, affinity chromatography, heparinase depolymerization), was shown to be a powerful tool providing relevant information about heparin structural peculiarities. The applied approach provided direct evidence that oligomers bearing glucuronic acid–glucosamine-3-O-sulfate at their nonreducing end represent an important structural signature of dalteparin. When extended to ATBR-related tetramers of bovine heparin, the heparanase-based approach allowed for elucidation of the structure of minor sequences that have not been reported yet. The obtained results are of high importance in the view of the growing interest of regulatory agencies and manufacturers in the development of low-molecular-weight heparin generics as well as bovine heparin as alternative source. MDPI 2019-12-02 /pmc/articles/PMC6930493/ /pubmed/31810297 http://dx.doi.org/10.3390/molecules24234403 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Alekseeva, Anna
Urso, Elena
Mazzini, Giulia
Naggi, Annamaria
Heparanase as an Additional Tool for Detecting Structural Peculiarities of Heparin Oligosaccharides
title Heparanase as an Additional Tool for Detecting Structural Peculiarities of Heparin Oligosaccharides
title_full Heparanase as an Additional Tool for Detecting Structural Peculiarities of Heparin Oligosaccharides
title_fullStr Heparanase as an Additional Tool for Detecting Structural Peculiarities of Heparin Oligosaccharides
title_full_unstemmed Heparanase as an Additional Tool for Detecting Structural Peculiarities of Heparin Oligosaccharides
title_short Heparanase as an Additional Tool for Detecting Structural Peculiarities of Heparin Oligosaccharides
title_sort heparanase as an additional tool for detecting structural peculiarities of heparin oligosaccharides
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6930493/
https://www.ncbi.nlm.nih.gov/pubmed/31810297
http://dx.doi.org/10.3390/molecules24234403
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