Polypyrrole oligosaccharide array and surface plasmon resonance imaging for the measurement of glycosaminoglycan binding interactions.
In order to construct tools able to screen oligosaccharide-protein interactions, we have developed a polypyrrole-based oligosaccharide chip constructed via a copolymerization process of pyrrole and pyrrole-modified oligosaccharide. For our study, GAG (glycosaminoglycans) or GAG fragments, which are involved in many fundamental biological processes, were modified by the pyrrole moiety on their reducing end and then immobilized on the chip. The parallel binding events on the upperside of the surface can be simultaneously monitored and quantified in real time and without labeling by surface plasmon resonance imaging (SPRi). We show that electrocopolymerization of the oligosaccharide-pyrrole above a gold surface enables the covalent immobilization of multiple probes and the subsequent monitoring of their binding capacities using surface plasmon resonance imaging. Moreover, a biological application was made involving different GAG fragments and different proteins, including stromal cell-derived factor-1alpha (SDF-1alpha), interferon-gamma (IFN-gamma), and monoclonal antibody showing different affinity pattern.
Mercey
Emilie
E
CREAB, UMR 5819 (CEA, CNRS, UJF), INAC CEA Grenoble; 17, Rue des Martyrs 38054 Grenoble Cedex 9, France.
Sadir
Rabia
R
Maillart
Emmanuel
E
Roget
André
A
Baleux
Françoise
F
Lortat-Jacob
Hugues
H
Livache
Thierry
T
D054377:Chemokine CXCL12 / Q000032:analysis; D006025:Glycosaminoglycans / Q000032:analysis*; D006493:Heparin / Q000032:analysis; D007371:Interferon-gamma / Q000032:analysis; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D009844:Oligosaccharides / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D011506:Proteins / Q000032:analysis*; D011758:Pyrroles / Q000737:chemistry*; D020349:Surface Plasmon Resonance / Q000379:methods*
D054377:Chemokine CXCL12; D006025:Glycosaminoglycans; D009844:Oligosaccharides; D011108:Polymers; D011506:Proteins; D011758:Pyrroles; C067635:polypyrrole; D007371:Interferon-gamma; D006493:Heparin