一种基于己糖激酶的新型电导生物传感器用于三磷酸腺苷的测定。
A novel conductometric biosensor based on hexokinase for determination of adenosine triphosphate.
作者信息
Kucherenko I S, Kucherenko D Yu, Soldatkin O O, Lagarde F, Dzyadevych S V, Soldatkin A P
机构信息
Institute of Molecular Biology and Genetics of National Academy of Sciences of Ukraine, 150 Zabolotny Str., Kyiv 03680, Ukraine; Taras Shevchenko National University of Kyiv, 64 Volodymyrska str., 01003 Kyiv, Ukraine; Institute of Analytical Sciences, UMR5280 CNRS/UCBL/ENS, 5 rue de la Doua, 69100, Villeurbanne, France.
Institute of Molecular Biology and Genetics of National Academy of Sciences of Ukraine, 150 Zabolotny Str., Kyiv 03680, Ukraine; Taras Shevchenko National University of Kyiv, 64 Volodymyrska str., 01003 Kyiv, Ukraine.
出版信息
Talanta. 2016 Apr 1;150:469-75. doi: 10.1016/j.talanta.2015.12.069. Epub 2015 Dec 28.
The paper presents a simple and inexpensive reusable biosensor for determination of the concentration of adenosine-5'-triphosphate (ATP) in aqueous samples. The biosensor is based on a conductometric transducer which contains two pairs of gold interdigitated electrodes. An enzyme hexokinase was immobilized onto one pair of electrodes, and bovine serum albumin-onto another pair (thus, a differential mode of measurement was used). Conditions of hexokinase immobilization on the transducer by cross-linking via glutaraldehyde were optimized. Influence of experimental conditions (concentration of magnesium ions, ionic strength and concentration of the working buffer) on the biosensor work was studied. The reproducibility of biosensor responses and operational stability of the biosensor were checked during one week. Dry storage at -18 °C was shown to be the best conditions to store the biosensor. The biosensor was successfully applied for measurements of ATP concentration in pharmaceutical samples. The proposed biosensor may be used in future for determination of ATP and/or glucose in water samples.
本文介绍了一种简单且成本低廉的可重复使用生物传感器，用于测定水性样品中腺苷 - 5'-三磷酸（ATP）的浓度。该生物传感器基于一个包含两对金叉指电极的电导式传感器。将己糖激酶固定在一对电极上，牛血清白蛋白固定在另一对电极上（因此，采用了差分测量模式）。优化了通过戊二醛交联将己糖激酶固定在传感器上的条件。研究了实验条件（镁离子浓度、离子强度和工作缓冲液浓度）对生物传感器工作的影响。在一周内检查了生物传感器响应的重现性和生物传感器的操作稳定性。结果表明，在 -18°C 下干燥储存是保存生物传感器的最佳条件。该生物传感器已成功应用于药物样品中 ATP 浓度的测量。所提出的生物传感器未来可用于测定水样中的 ATP 和/或葡萄糖。
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