Laboratory of Biomolecular Electronics, Institute of Molecular Biology and Genetics, NAS of Ukraine, 150 Zabolotnogo str., Kyiv 03148, Ukraine; Institute of High Technologies, Taras Shevchenko National University of Kyiv, 64, Volodymyrska Str., Kyiv 01003, Ukraine.
Laboratory of Biomolecular Electronics, Institute of Molecular Biology and Genetics, NAS of Ukraine, 150 Zabolotnogo str., Kyiv 03148, Ukraine; Department of Mechanical Engineering, Iowa State University, Ames, IA 50011, United States.
Bioelectrochemistry. 2019 Aug;128:100-108. doi: 10.1016/j.bioelechem.2019.03.010. Epub 2019 Mar 31.
The work was aimed at the development of a biosensor array for the simultaneous determination of six solutes (glutamate, glucose, choline, acetylcholine, lactate, and pyruvate) in aqueous solutions. Enzymes selective for these substrates were immobilized on the surface of amperometric platinum disc electrodes and served as bioselective elements of a biosensor array. Direct enzymatic analysis by the developed biosensors provided high sensitivity to the tested substrates (limits of detection were 1-5 μM). The linear ranges of the biosensors were from 0.001-0.01 mM to 0.2-2.5 mM. The influence of solution pH, ionic strength and buffer capacity on the biosensor responses was investigated; the conditions for simultaneous operation of all the bioselective elements were optimized. The absence of any cross-influence of the substrates of enzymatic systems used was shown as well as a high selectivity of the biosensors and the absence of any impact of interfering substances (ascorbic acid, dopamine, cysteine, paracetamol). The developed biosensor array had good response reproducibility and storage stability. The array is suitable for rapid (0.5-1 min) and simple simultaneous determination of glutamate, glucose, choline, acetylcholine, lactate, and pyruvate in aqueous (biological) samples; furthermore, the creation of a single chip with six sensitive elements is possible as well as the addition of other biosensors.
本工作旨在开发一种用于同时测定水溶液中六种溶质（谷氨酸、葡萄糖、胆碱、乙酰胆碱、乳酸和丙酮酸）的生物传感器阵列。对这些底物具有选择性的酶固定在电流测定型铂圆盘电极的表面上，作为生物传感器阵列的生物选择性元件。通过所开发的生物传感器进行的直接酶分析对测试底物具有高灵敏度（检测限为 1-5μM）。生物传感器的线性范围从 0.001-0.01mM 到 0.2-2.5mM。研究了溶液 pH、离子强度和缓冲能力对生物传感器响应的影响；优化了所有生物选择性元件同时操作的条件。结果表明，所使用的酶系统的底物之间没有任何交叉影响，生物传感器具有高选择性，并且没有任何干扰物质（抗坏血酸、多巴胺、半胱氨酸、对乙酰氨基酚）的影响。所开发的生物传感器阵列具有良好的响应重现性和存储稳定性。该阵列适用于快速（0.5-1 分钟）和简单同时测定水溶液（生物）样品中的谷氨酸、葡萄糖、胆碱、乙酰胆碱、乳酸和丙酮酸；此外，有可能创建具有六个敏感元件的单个芯片，并且可以添加其他生物传感器。
