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基于新型铵离子选择性铜-聚苯胺纳米复合材料的肌氨酸酐和尿素生物传感器。

Creatinine and urea biosensors based on a novel ammonium ion-selective copper-polyaniline nano-composite.

机构信息

Laboratory of Biomolecular Electronics, Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine, 03680 Kyiv, Ukraine; Biosensors & Bioelectronics Centre, IFM, Linköping University, 58183, Sweden.

Biosensors & Bioelectronics Centre, IFM, Linköping University, 58183, Sweden.

出版信息

Biosens Bioelectron. 2016 Mar 15;77:505-11. doi: 10.1016/j.bios.2015.10.009. Epub 2015 Oct 9.

Abstract

The use of a novel ammonium ion-specific copper-polyaniline nano-composite as transducer for hydrolase-based biosensors is proposed. In this work, a combination of creatinine deaminase and urease has been chosen as a model system to demonstrate the construction of urea and creatinine biosensors to illustrate the principle. Immobilisation of enzymes was shown to be a crucial step in the development of the biosensors; the use of glycerol and lactitol as stabilisers resulted in a significant improvement, especially in the case of the creatinine, of the operational stability of the biosensors (from few hours to at least 3 days). The developed biosensors exhibited high selectivity towards creatinine and urea. The sensitivity was found to be 85 ± 3.4 mAM(-1)cm(-2) for the creatinine biosensor and 112 ± 3.36 mAM(-1)cm(-2) for the urea biosensor, with apparent Michaelis-Menten constants (KM,app), obtained from the creatinine and urea calibration curves, of 0.163 mM for creatinine deaminase and 0.139 mM for urease, respectively. The biosensors responded linearly over the concentration range 1-125 µM, with a limit of detection of 0.5 µM and a response time of 15s. The performance of the biosensors in a real sample matrix, serum, was evaluated and a good correlation with standard spectrophotometric clinical laboratory techniques was found.

摘要

本文提出了一种新型铵离子特异性铜-聚苯胺纳米复合材料作为基于水解酶的生物传感器的换能器。在这项工作中,选择肌酸脱氨酶和脲酶的组合作为模型系统,以展示构建尿素和肌酸生物传感器的原理。酶的固定化被证明是生物传感器发展的关键步骤;使用甘油和山梨醇作为稳定剂可显著提高生物传感器的操作稳定性(从几个小时至少提高到 3 天),特别是在肌酸的情况下。所开发的生物传感器对肌酸和尿素表现出高选择性。研究发现,肌酸生物传感器的灵敏度为 85 ± 3.4 mAM(-1)cm(-2),尿素生物传感器的灵敏度为 112 ± 3.36 mAM(-1)cm(-2),从肌酸脱氨酶和脲酶的肌酐和尿素校准曲线中获得的表观米氏常数(KM,app)分别为 0.163 mM 和 0.139 mM。生物传感器在 1-125 µM 的浓度范围内呈线性响应,检测限为 0.5 µM,响应时间为 15s。在实际样本基质血清中评估了生物传感器的性能,发现与标准分光光度临床实验室技术具有良好的相关性。

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