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用于谷氨酸测量的酶修饰微电极:表征与应用

Enzyme-modified microelectrodes for measurement of glutamate: Characterization and applications.

作者信息

Alyamni Nadiah, Cook Clarice, Abot Jandro L, Zestos Alexander G

机构信息

Department of Chemistry, American University, Washington, D.C.

Department of Biomedical Engineering, Catholic University, Washington, D.C.

出版信息

Electroanalysis. 2025 Mar;37(3). doi: 10.1002/elan.12041. Epub 2025 Mar 17.

Abstract

Glutamate is a critical neurotransmitter in the central nervous system that plays a key role in numerous physiological processes and neurological disorders. Traditional methods of glutamate detection have low spatiotemporal resolution, while electrochemical methods are limited due to glutamate not being readily redox active at unmodified carbon electrode surfaces. This study presents the development of a glutamate oxidase-modified microelectrode for the sensitive, real-time detection of glutamate using fast-scan cyclic voltammetry (FSCV) with a triangle waveform. Here, we employed a chitosan-hydrogel coating to immobilize glutamate oxidase onto carbon fiber microelectrodes, enabling selective metabolism of glutamate to hydrogen peroxide. The metabolism to hydrogen peroxide facilitates indirect detection with high sensitivity across a concentration range relevant to physiological concentrations. We utilized FSCV for detection, which enhanced temporal resolution and chemical selectivity, allowing for the co-detection of glutamate with other neurotransmitters such as dopamine and norepinephrine. We performed proof-of-concept validation and testing utilizing both biological fluids and complex food samples, demonstrating the enzyme-modified microelectrode's broad applicability in clinical diagnostics and food quality assessment. The sensor showed excellent stability, resistance to fouling, and retained over 90% of its initial response after multiple uses. This work highlights the potential of this biosensor as a versatile tool for minimally invasive, biocompatible, rapid, and accurate glutamate measurement in a wide variety of samples for a diverse set of applications.

摘要

谷氨酸是中枢神经系统中的一种关键神经递质,在众多生理过程和神经疾病中发挥着关键作用。传统的谷氨酸检测方法时空分辨率较低,而电化学方法由于谷氨酸在未修饰的碳电极表面不易发生氧化还原反应而受到限制。本研究介绍了一种用于谷氨酸氧化酶修饰的微电极的开发,该微电极使用三角波快速扫描循环伏安法(FSCV)对谷氨酸进行灵敏、实时检测。在此,我们采用壳聚糖水凝胶涂层将谷氨酸氧化酶固定在碳纤维微电极上,使谷氨酸选择性代谢为过氧化氢。代谢为过氧化氢有助于在与生理浓度相关的浓度范围内进行高灵敏度的间接检测。我们利用FSCV进行检测,提高了时间分辨率和化学选择性,从而能够与多巴胺和去甲肾上腺素等其他神经递质共同检测谷氨酸。我们使用生物体液和复杂的食品样品进行了概念验证和测试,证明了酶修饰微电极在临床诊断和食品质量评估中的广泛适用性。该传感器表现出优异的稳定性、抗污染性,并且在多次使用后仍保留了超过90%的初始响应。这项工作突出了这种生物传感器作为一种通用工具的潜力,可用于在各种样品中进行微创、生物相容、快速且准确的谷氨酸测量,以满足多种应用需求。

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