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基于吡咯并喹啉醌(PQQ)-葡萄糖脱氢酶的荧光传感器,用于通过适配智能手机的信号分析进行葡萄糖检测。

Fluorescent sensor based on pyrroloquinoline quinone (PQQ)-glucose dehydrogenase for glucose detection with smartphone-adapted signal analysis.

作者信息

Wells Paulina K, Melman Artem, Katz Evgeny, Smutok Oleh

机构信息

Department of Chemistry and Biomolecular Science, Clarkson University, Potsdam, NY, 13699, USA.

出版信息

Mikrochim Acta. 2022 Sep 6;189(10):371. doi: 10.1007/s00604-022-05466-4.

Abstract

A new nano-structured platform for fluorescent analysis using PQQ-dependent glucose dehydrogenase (PQQ-GDH) was developed, particularly using a smartphone for transduction and quantification of optical signals. The PQQ-GDH enzyme was immobilized on SiO nanoparticles deposited on glass microfiber filter paper, providing a high load of the biocatalytic enzyme. The platform was tested and optimized for glucose determination using a wild type of the PQQ-GDH enzyme. The analysis was based on the fluorescence generated by the reduced form of phenazine methosulfate produced stoichiometrically to the glucose concentration. The fluorescent signals were generated at separate analytical spots on the paper support under wavelength (365 nm) UV excitation. The images of the analytical spots, dependent on the glucose concentration, were obtained using a photo camera of a standard smartphone. Then, the images were processed and quantified using software installed in a smartphone. The developed biocatalytic platform is the first step to assembling a large variety of biosensors using the same platform functionalized with artificial allosteric chimeric PQQ-dependent glucose dehydrogenase activated with different analytes. The future combination of the artificial enzymes, the presently developed analytical platform, and signal processing with a smartphone will lead to novel point-of-care and end-user biosensors applicable to virtually all possible analytes.

摘要

开发了一种使用依赖于吡咯喹啉醌的葡萄糖脱氢酶(PQQ-GDH)进行荧光分析的新型纳米结构平台,特别是使用智能手机来传导和定量光信号。将PQQ-GDH酶固定在沉积在玻璃微纤维滤纸上的SiO纳米颗粒上,从而提供高负载的生物催化酶。使用野生型PQQ-GDH酶对该平台进行了葡萄糖测定测试和优化。该分析基于与葡萄糖浓度化学计量产生的硫酸甲酯吩嗪还原形式产生的荧光。在波长(365 nm)紫外线激发下,在纸质载体上的单独分析点产生荧光信号。使用标准智能手机的摄像头获取取决于葡萄糖浓度的分析点图像。然后,使用安装在智能手机中的软件对图像进行处理和定量。所开发的生物催化平台是使用由不同分析物激活的人工变构嵌合PQQ依赖性葡萄糖脱氢酶功能化的同一平台组装各种生物传感器的第一步。未来人工酶、目前开发的分析平台以及智能手机信号处理的结合将产生适用于几乎所有可能分析物的新型即时护理和终端用户生物传感器。

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