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

机译:聚吡咯寡糖阵列和表面等离振子共振成像用于测量糖胺聚糖结合相互作用

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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.
机译:为了构建能够筛选寡糖蛋白质相互作用的工具,我们开发了一种通过吡咯和吡咯修饰的寡糖的共聚过程构建的基于聚吡咯的寡糖芯片。在我们的研究中,涉及许多基本生物学过程的GAG(糖胺聚糖)或GAG片段在其还原端被吡咯部分修饰,然后固定在芯片上。可以同时实时监控和定量表面上侧的平行结合事件,而无需通过表面等离振子共振成像(SPRi)进行标记。我们表明,金表面上的寡糖吡咯的电共聚使多个探针能够共价固定,并随后使用表面等离子体共振成像监测其结合能力。此外,进行了涉及不同GAG片段和不同蛋白质的生物学应用,包括基质细胞衍生因子1α(SDF-1 alpha),干扰素-γ(IFN-γ)和显示不同亲和力模式的单克隆抗体。

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