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一种用于选择性检测活细胞中 K⁺诱导释放的多巴胺的安培型纳米生物传感器。

An amperometric nanobiosensor for the selective detection of K⁺-induced dopamine released from living cells.

机构信息

Department of Chemistry, Institute of Biophysio Sensor Technology (IBST), Pusan National University, Busan 609-735, South Korea.

Department of Internal Medicine, Lee Gil Ya Cancer and Diabetes Institute, Gachon University of Medicine and Science, Incheon, South Korea.

出版信息

Biosens Bioelectron. 2015 Jun 15;68:421-428. doi: 10.1016/j.bios.2015.01.024. Epub 2015 Jan 12.

Abstract

A highly sensitive amperometric sensor has been studied for selective monitoring of K(+)-induced dopamine released from dopaminergic cells (PC12) which is based on an EDTA immobilized-poly(1,5-diaminonaphthalne) (poly-DAN) layer comprising graphene oxide (GO) and gold nanoparticles (GO/AuNPs). The integration of a negatively charged probe molecule on the poly-DAN/GO/AuNPs nanohybrid attained the signal enhancement to discriminate dopamine (DA) molecules from foreign species by catalytic effect and surface charge, and hydrogen bonding-based interactions with a probe molecule. The sensor performance and morphology were investigated using voltammetry, impedance spectrometry, SEM, and XPS. Experimental variables affecting the analytical performance of the sensor probe were optimized, and linear response was observed in the range of 10 nM-1 µM with a detection limit of 5.0 nM (±0.01) for DA. Then, the sensor was applied to monitor dopamine released from PC12 cells upon extracellular stimulation of K(+) ions. It was also confirmed that K(+)-induced dopamine release was inhibited by a calcium channel inhibitor (Nifidipine). The results demonstrated that the presented biosensor could be used as an excellent tool for monitoring the effect of exogenous agents on living cells and drug efficacy tests.

摘要

研究了一种基于包含氧化石墨烯(GO)和金纳米粒子(GO/AuNPs)的乙二胺四乙酸固定化-聚(1,5-二氨基萘)(poly-DAN)层的高灵敏度电流传感器,用于选择性监测多巴胺能细胞(PC12)中 K(+)-诱导的多巴胺释放。聚-DAN/GO/AuNPs 纳米杂化体上带负电荷的探针分子的整合通过催化作用和表面电荷以及与探针分子的氢键相互作用,实现了信号增强,从而可以区分多巴胺(DA)分子和外来物质。使用伏安法、阻抗谱法、SEM 和 XPS 研究了传感器的性能和形态。优化了影响传感器探针分析性能的实验变量,并观察到在 10 nM-1 µM 范围内呈线性响应,检测限为 5.0 nM(±0.01)。然后,将传感器用于监测 K(+)离子细胞外刺激后 PC12 细胞中多巴胺的释放。还证实钙通道抑制剂(硝苯地平)抑制 K(+)-诱导的多巴胺释放。结果表明,该生物传感器可作为监测外源性药物对活细胞的影响和药物疗效测试的有效工具。

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