Polysaccharide microarrays for polysaccharide/growth factor interaction studies

Clement L. Roux 1,3Remi Desmet 1Latifa Dahri-Correia 2Hartmut F. Hildebrand 3Oleg Melnyk 1

1. Faculté de Médecine (GRB), 1, place de Verdun, Lille 59045, France
2. Biodex, 637, rue des Aulnois, Saint Amand-les-Eaux 59230, France
3. Institut de Biologie de Lille (IBL), 1 rue du professeur Calmette, Lille 59021, France

Abstract

Sulfated polysaccharides are known to potentiate the in-vitro activity of certain proteins, such as growth factors. We have in our hands a library of 115 dextrans functionalised with various amounts of carboxymethyl, benzylamide and sulfate moieties (DMCBSu, Dextran methylcarboxylate benzylamide sulfate)1 that mimic the properties of heparin. In order to identify rapidly sets of compounds that bind growth factors, we developed microarrays on which these large polymers (above 40 kDa) are printed and immobilised by physisorption.

The library of DMCBSu is designed in such a way that it gives us an insight into which substitution patterns are most favorable for the interactions with given proteins. This identification method has already been validated for interactions with platelet derived growth factor-BB2. In order to evaluate the affinity of each dextrans with Bone morphogenetic protein-23, a growth factor involved in osteoinduction and bone regeneration, microarrays were incubated with a solution of the cytokine. Binding intensities were determined with a fluorescence-based detection technique, through consecutive incubations with biotinylated anti-BMP2, and Cy3-labelled streptavidin. After semi-automated data analysis, this strategy enabled us to isolate groups of compounds with different affinities for BMP2, which could be employed in various biomedical applications. Compounds isolated in such a way will soon be tested for their ability to potentiate the action of BMP-2 in-vitro.

1 Huynh et al, Carbohydr Res. 2001, 332, 75-83.

2 Carion et al, Chembiochem. 2006, 7, 817-826.

3 Chen et al, Growth Factors. 2004, 4, 233-41.

 

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Presentation: Poster at E-MRS Fall Meeting 2006, Symposium J, by Clement L. Roux
See On-line Journal of E-MRS Fall Meeting 2006

Submitted: 2006-06-08 08:13
Revised:   2009-06-07 00:44