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葡萄糖氧化酶固定于聚苯乙烯-嵌段-聚(2-乙烯基吡啶)纳米结构薄膜上。

Immobilization of glucose oxidase to nanostructured films of polystyrene-block-poly(2-vinylpyridine).

作者信息

Bhakta Samir A, Benavidez Tomas E, Garcia Carlos D

机构信息

Department of Chemistry, The University of Texas at San Antonio One UTSA Circle, San Antonio, TX 78249, USA.

Department of Chemistry, The University of Texas at San Antonio One UTSA Circle, San Antonio, TX 78249, USA.

出版信息

J Colloid Interface Sci. 2014 Sep 15;430:351-6. doi: 10.1016/j.jcis.2014.05.067. Epub 2014 Jun 12.

Abstract

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, 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.

摘要

生物传感器开发的关键步骤之一是将生物识别元件固定在基底表面。在可作为基底的其他材料中,嵌段共聚物在蛋白质固定方面具有尚未开发的潜力,能提供显著优势。为探索这种可能性,本论文描述了聚苯乙烯 - 嵌段 - 聚(2 - 乙烯基吡啶)(PS - b - P2VP)薄膜的制备与表征。然后用这些薄膜研究葡萄糖氧化酶的固定,葡萄糖氧化酶是生物传感器开发中的一种模型酶。根据所呈现的结果,纳米多孔薄膜能显著增加基底的表面积,并固定大量活性酶。论文中讨论的基底 - 酶界面表征旨在提供有关表面(材料、几何形状和孔隙密度)、蛋白质结构以及提高酶的催化活性和稳定性所需的固定条件(pH值和蛋白质浓度)之间关系的关键信息。PS - b - P2VP纳米多孔薄膜实现的最大归一化活性为3300±700 U m(-2)。

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