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Open AccessFeature PaperArticle

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

by Anna Alekseeva 1,*,†, Elena Urso 2,*,†, Giulia Mazzini 1,‡ and Annamaria Naggi 1,2
1
Centro Alta Tecnologia Istituto di Ricerche Chimiche e Biochimiche G. Ronzoni Srl, via G. Colombo 81, 20133 Milan, Italy
2
Istituto di Ricerche Chimiche e Biochimiche G. Ronzoni, via G. Colombo 81, 20133 Milan, Italy
*
Authors to whom correspondence should be addressed.
Authors contributed equally to this work.
Present address: IRCCS Policlinico San Matteo, viale Golgi 19, 27100 Pavia, Italy; [email protected] (G.M.)
Academic Editors: Giangiacomo Torri, Jawed Fareed and Job Harenberg
Molecules 2019, 24(23), 4403; https://doi.org/10.3390/molecules24234403
Received: 13 November 2019 / Revised: 29 November 2019 / Accepted: 30 November 2019 / Published: 2 December 2019
(This article belongs to the Special Issue Emerging Frontiers in Glycosaminoglycans and Mimetics)
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. View Full-Text
Keywords: heparanase; heparin lyases; low molecular weight heparins; bovine mucosal heparin; antithrombin binding site; mass spectrometry heparanase; heparin lyases; low molecular weight heparins; bovine mucosal heparin; antithrombin binding site; mass spectrometry
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Alekseeva, A.; Urso, E.; Mazzini, G.; Naggi, A. Heparanase as an Additional Tool for Detecting Structural Peculiarities of Heparin Oligosaccharides. Molecules 2019, 24, 4403.

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