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基于 GOx/HRP 的裸眼比色葡萄糖生物传感器的设计:稳定的静电纺丝共混壳聚糖-聚乙烯醇纳米纤维的制备。

Fabrication of stable electrospun blended chitosan-poly(vinyl alcohol) nanofibers for designing naked-eye colorimetric glucose biosensor based on GOx/HRP.

机构信息

Yıldız Technical University, Metallurgy and Materials Engineering Department, İstanbul 34210, Turkey.

Yıldız Technical University, Bioengineering Department, İstanbul 34210, Turkey.

出版信息

Int J Biol Macromol. 2021 Dec 1;192:999-1012. doi: 10.1016/j.ijbiomac.2021.10.048. Epub 2021 Oct 13.

Abstract

In this study, designing of a stable electrospun blended chitosan (CS)-poly(vinyl alcohol) (PVA) nanofibers for colorimetric glucose biosensing in an aqueous medium was investigated. CS and PVA solutions were blended to acquire an optimum content (CS/PVA:1/4) and electrospunned to obtain uniform and bead-free CS/PVA nanofiber structures following the optimization of the electrospinning parameters (33 kV, 20 cm, and 1.2 ml.h). Crosslinking process applied subsequently provided mechanically and chemically stable nanofibers with an average diameter of 378 nm. The morphological homogeneity, high fluid absorption ability (>%50), thermal (<230 °C) and morphological stability, surface hydrophilicity and degrability properties of cross-linked CS/PVA nanofiber demonstrated their great potential to be developed as an eye-readable strip for biosensing applications. The glucose oxidase (GOx) and horseradish peroxidase (HRP) was immobilized by physical adsorption on the cross-linked CS/PVA nanofiber. The glucose assay analysis by ultraviolet-visible (UV-Vis) spectrophotometry using the same enzymatic system of the proposed glucose strips in form of absorbance versus concentration plot was found to be linear over a glucose concentration range of 2.7 to 13.8 mM. The prepared naked eye colorimetric glucose detection strips, with lower detection limit of 2.7 mM, demonstrated dramatic color change from white (0 mM) to brownish-orange (13.8 mM). The developed cross-linked CS/PVA nanofiber strips, prepared by electrospinnig procedure, could be easily adapted to a color map, as an alternative material for glucose sensing. Design of a practical, low-cost, and environmental-friendly bio-based CS/PVA testing strips for eye readable detection were presented and suggested as an applicable medium for a wide range of glucose concentrations.

摘要

在这项研究中,设计了一种稳定的电纺共混壳聚糖(CS)-聚乙烯醇(PVA)纳米纤维,用于在水介质中进行比色葡萄糖生物传感。混合 CS 和 PVA 溶液以获得最佳含量(CS/PVA:1/4),并通过优化电纺参数(33 kV、20 cm 和 1.2 ml.h)获得均匀且无珠的 CS/PVA 纳米纤维结构。随后进行的交联过程提供了机械和化学稳定的纳米纤维,平均直径为 378 nm。交联 CS/PVA 纳米纤维的形态均匀性、高流体吸收能力(>50%)、热(<230°C)和形态稳定性、表面亲水性和可降解性特性表明,它们具有作为生物传感应用的眼可读条带开发的巨大潜力。葡萄糖氧化酶(GOx)和辣根过氧化物酶(HRP)通过物理吸附固定在交联的 CS/PVA 纳米纤维上。通过使用相同的酶系统,通过紫外-可见(UV-Vis)分光光度法对葡萄糖进行分析,以吸收度与浓度的关系图表示,在 2.7 至 13.8 mM 的葡萄糖浓度范围内呈线性关系。制备的裸眼比色葡萄糖检测条具有较低的检测限 2.7 mM,显示出从白色(0 mM)到棕橙色(13.8 mM)的剧烈颜色变化。通过静电纺丝工艺制备的交联 CS/PVA 纳米纤维条可以很容易地适应比色图谱,作为葡萄糖传感的替代材料。提出了一种实用、低成本、环保的基于生物的 CS/PVA 测试条的设计,用于肉眼可读的检测,并建议将其作为广泛葡萄糖浓度的适用介质。

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