Please use this identifier to cite or link to this item: https://repositorio.unifesp.br/handle/11600/30737
Title: Development of an enzyme-linked immunosorbent assay (ELISA)-like fluorescence assay to investigate the interactions of glycosaminoglycans to cells
Authors: Boucas, Rodrigo Ippolito [UNIFESP]
Trindade, Edvaldo S. [UNIFESP]
Tersariol, Ivarne L. S. [UNIFESP]
Dietrich, Carl P. [UNIFESP]
Nader, Helena B. [UNIFESP]
Universidade Federal de São Paulo (UNIFESP)
Univ Fed Parana
Univ Mogi das Cruzes
Keywords: glycosaminoglycans
biotinylation procedures
fluorescence
endothelial cells
smooth muscle cells
Issue Date: 23-Jun-2008
Publisher: Elsevier B.V.
Citation: Analytica Chimica Acta. Amsterdam: Elsevier B.V., v. 618, n. 2, p. 218-226, 2008.
Abstract: Sulfated glycosaminoglycans were labeled with biotin to study their interaction with cells in culture. Thus, heparin, heparan sulfate, chondroitin 4-sulfate, chondroitin 6-sulfate and dermatan sulfate were labeled using biotin-hydrazide, under different conditions. the structural characteristics of the biotinylated products were determined by chemical (molar ratios of hexosamine, uronic acid, sulfate and biotin) and enzymatic methods (susceptibility to degradation by chondroitinases and heparitinases). the binding of biotinylated glycosaminoglycans was investigated both in endothelial and smooth muscle cells in culture, using a novel time resolved fluorometric method based on interaction of europium-labeled streptavidin with the biotin covalently linked to the compounds. the interactions of glycosaminoglycans were saturable and number of binding sites could be obtained for each individual compound. the apparent dissociation constant varied among the different glycosaminoglycans and between the two cell lines, the interactions of the biotinylated glycosaminoglycans with the cells were also evaluated using confocal microscopy. We propose a convenient and reliable method for the preparation of biotinylated glycosaminoglycans, as well as a sensitive non-competitive fluorescence-based assay for studies of the interactions and binding of these compounds to cells in culture. (c) 2008 Elsevier B.V. All rights reserved.
URI: http://repositorio.unifesp.br/handle/11600/30737
ISSN: 0003-2670
Other Identifiers: http://dx.doi.org/10.1016/j.aca.2008.04.059
Appears in Collections:Em verificação - Geral

Files in This Item:
There are no files associated with this item.


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.