Thesis presented November 17, 2005

Abstract:
The thesis goal is to design an oligosaccharide chip, compatible with surface plasmon resonance imaging process, where parallel binding events occurring on the top of the surface can be simultaneously monitored and quantified in real time. A model system using heparin as sugar to immobilize has been proposed. The modification of the sugar by a pyrrole group allows to immobilize the oligosaccharide covalently by an electrochemical process. Thus, different molecules containing a pyrrole group and a linker were synthesized. Then the chip was made by successive electrocopolymerizations. The spots can be analyzed by SEM and AFM, before testing the biological recognition between the modified oligosaccharides and the protein. The first experiments using fluorescent labels have allowed to validate the developed system and the suggested chemistry: the presence of negative spots confirm the specificity of the biological interactions detected by fluorescence. Thus, SPR imaging allows to confirm biological interactions sensitivity. Because of the measurements in real time, kinetics of association and dissociation phases can be studied. Following the optimization of the chemistry, this approach was finally applied to the study of different biological problems.

Keywords:
oligosaccharide interactions, protein, electrocopolymerization, SPR imaging, surface plasmon resonance

On-line thesis.