Polypyrrole oligosaccharide array and surface plasmon resonance imaging for the measurement of glycosaminoglycan binding interaction

被引:57
|
作者
Mercey, Emilie [1 ]
Sadir, Rabia [2 ]
Maillart, Emmanuel [3 ]
Roget, Andre [1 ]
Baleux, Francoise [4 ]
Lortat-Jacob, Hugues [2 ]
Livache, Thierry [1 ]
机构
[1] INAC CEA Grenoble, CREAB, CEA,UMR 5819, CNRS,UJF, F-38054 Grenoble 9, France
[2] CEA, CNRS, UJF, Inst Biol Struct,UMR 5075, F-38027 Grenoble 1, France
[3] Genopt SA, Orsay, France
[4] Inst Pasteur, F-75015 Paris, France
关键词
D O I
10.1021/ac800226k
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
In order to construct tools able to screen oligosaccharideprotein interactions, we have developed a polypyrrolebased 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-1 alpha (SDF-1 alpha), interferon-gamma (IFN-gamma), and monoclonal antibody showing different affinity pattern.
引用
收藏
页码:3476 / 3482
页数:7
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