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Immobilization of Glucose Oxidase to Nanostructured Films of Polystyrene-block-poly(2-vinylpyridine)

机译:葡萄糖氧化酶固定到聚苯乙烯嵌段聚(2-乙烯基吡啶)的纳米结构膜上

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摘要

A critical step for the development of biosensors is the immobilization of the biorecognition element to the surface of a substrate. Among other materials that can be used as substrates, block copolymers have the untapped potential to provide significant advantages for the immobilization of proteins. To explore such possibility, this manuscript describes the fabrication and characterization of thin-films of polystyrene-block-poly(2-vinylpyridine) (PS-b-P2VP). These films were then used to investigate the immobilization of glucose oxidase, a model enzyme for the development of biosensors. According to the results presented, the nanoporous films can provide significant increases in surface area of the substrate and the immobilization of larger amounts of active enzyme. The characterization of the substrate-enzyme interface discussed in the manuscript aims to provide critical information about relationship between the surface (material, geometry, and density of pores), the protein structure, and the immobilization conditions (pH, ionic strength, and protein concentration) required to improve the catalytic activity and stability of the enzymes. A maximum normalized activity of 3300 ± 700 U m−2 was achieved for the nanoporous film of PS-b-P2VP.
机译:开发生物传感器的关键步骤是将生物识别元件固定在基质表面上。在可用作底物的其他材料中,嵌段共聚物具有未开发的潜力,可为固定蛋白质提供显着的优势。为了探索这种可能性,该手稿描述了聚苯乙烯嵌段聚(2-乙烯基吡啶)(PS-b-P2VP)薄膜的制造和表征。然后将这些薄膜用于研究葡萄糖氧化酶的固定化,葡萄糖氧化酶是用于开发生物传感器的模型酶。根据给出的结果,纳米多孔膜可以显着增加底物的表面积并固定大量的活性酶。本文中讨论的底物-酶界面的特征旨在提供有关表面(材料,几何形状和孔的密度),蛋白质结构和固定条件(pH,离子强度和蛋白质浓度)之间关系的关键信息。 ),以提高酶的催化活性和稳定性。 PS-b-P2VP纳米多孔膜的最大归一化活性为3300±700 U m −2

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