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首页> 外文期刊>The Analyst: The Analytical Journal of the Royal Society of Chemistry: A Monthly International Publication Dealing with All Branches of Analytical Chemistry >Graphene based electrochemical biosensor for label-free measurement of the activity and inhibition of protein tyrosine kinase
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Graphene based electrochemical biosensor for label-free measurement of the activity and inhibition of protein tyrosine kinase

机译：基于石墨烯的电化学生物传感器，用于无标记测量蛋白酪氨酸激酶的活性和抑制

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摘要

In this paper, we report a simple, ultrasensitive and label-free method to evaluate the activity of protein tyrosine (Tyr) kinase based on the electrochemical signal of Tyr residues at a graphene modified glassy carbon electrode. It was found that graphene could enhance the electrochemical response of Tyr through electrocatalytic oxidation reaction. After phosphorylation by kinase, the phosphorylated Tyr (pTyr) is electro-inactive and the electrochemical signal is reduced. Therefore, the electrochemical response of Tyr residues in peptides can be used as a signal reporter to assay kinase activity. In this study, using Src-catalyzed Tyr-phosphorylation as a model, the activity of kinase can be evaluated with a linear range from 0.26 to 33.79 nM and an extraordinarily low detection limit of 0.087 nM. Moreover, this electrochemical biosensor can also be utilized for monitoring the inhibition of kinase using 4-amino-5-(4-chlorophenyl)-7-(tert-butyl) pyrazolo [3,4-d] pyrimidine, a small molecule inhibitor. On the basis of the inhibitor concentration dependent Tyr oxidation signal, the IC_(50) value was estimated to be 99 nM.
机译：在本文中，我们报告了一种基于石墨烯修饰的玻碳电极上Tyr残基的电化学信号来评估蛋白酪氨酸（Tyr）激酶活性的简单，超灵敏且无标签的方法。发现石墨烯可以通过电催化氧化反应增强Tyr的电化学反应。通过激酶进行磷酸化后，磷酸化的Tyr（pTyr）失电，电化学信号降低。因此，肽中Tyr残基的电化学反应可用作信号报告子，以测定激酶活性。在这项研究中，使用Src催化的Tyr磷酸化作为模型，可以在0.26至33.79 nM的线性范围和0.087 nM的极低检测限范围内评估激酶的活性。此外，该电化学生物传感器还可以利用小分子抑制剂4-氨基-5-（4-氯苯基）-7-（叔丁基）吡唑并[3，4-d]嘧啶来监测激酶的抑制作用。根据抑制剂浓度依赖的Tyr氧化信号，IC_（50）值估计为99 nM。

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