Mercey Emilie, Sadir Rabia, Maillart Emmanuel, Roget André, Baleux Françoise, Lortat-Jacob Hugues, Livache Thierry
CREAB, UMR 5819 (CEA, CNRS, UJF), INAC CEA Grenoble; 17, Rue des Martyrs 38054 Grenoble Cedex 9, France.
Anal Chem. 2008 May 1;80(9):3476-82. doi: 10.1021/ac800226k. Epub 2008 Mar 19.
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.
为构建能够筛选寡糖-蛋白质相互作用的工具，我们开发了一种基于聚吡咯的寡糖芯片，该芯片通过吡咯与吡咯修饰的寡糖的共聚过程构建而成。在我们的研究中，参与许多基本生物学过程的糖胺聚糖（GAG）或GAG片段在其还原端通过吡咯部分进行修饰，然后固定在芯片上。表面上侧的平行结合事件可通过表面等离子体共振成像（SPRi）实时、无需标记地同时进行监测和定量。我们表明，在金表面上方进行寡糖-吡咯的电共聚能够实现多种探针的共价固定，并随后使用表面等离子体共振成像监测其结合能力。此外，还进行了一项生物学应用，涉及不同的GAG片段和不同的蛋白质，包括基质细胞衍生因子-1α（SDF-1α）、干扰素-γ（IFN-γ）以及显示出不同亲和力模式地单克隆抗体。
