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3D 双金属 Au/Pt 纳米花修饰的针型微电极用于活细胞分泌的 ATP 的直接原位监测。

3D bimetallic Au/Pt nanoflowers decorated needle-type microelectrode for direct in situ monitoring of ATP secreted from living cells.

机构信息

Biosensor National Special Laboratory, Key Laboratory of Biomedical Engineering of Ministry of Education, College of Biomedical Engineering and Instrument Science, Innovation Center for Minimally Invasive Technique and Device, Zhejiang University, Hangzhou, 310027, PR China.

Biosensor National Special Laboratory, Key Laboratory of Biomedical Engineering of Ministry of Education, College of Biomedical Engineering and Instrument Science, Innovation Center for Minimally Invasive Technique and Device, Zhejiang University, Hangzhou, 310027, PR China.

出版信息

Biosens Bioelectron. 2020 Apr 1;153:112019. doi: 10.1016/j.bios.2020.112019. Epub 2020 Jan 11.

Abstract

Adenosine triphosphate (ATP) plays a crucial role in energy metabolism and extracellular purinergic signaling. A 3D bimetallic Au/Pt nanoflowers decorated ATP microelectrode biosensor prepared by facile and effective template-free electrodeposition was firstly reported, realizing local detection of cellular ATP secretion. The ATP biosensor was developed by co-immobilization of glucose oxidase and hexokinase, exhibiting long-term stability (79.39 ± 9.15% of its initial value remained after 14 days at 4 °C) and high selectivity with a limit of detection down to 2.5 μM (S/N = 3). The resulting ATP biosensor was then used for direct in situ monitoring of ATP secreted from living cells (PC12) with the stimulation of high K solutions. The obtained current was about 21.6 ± 3.4 nA (N = 6), corresponding to 12.2 ± 2.8 μM ATP released from cells, right in the micromolar range and consistent with the suggested levels. The 3D bimetallic Au/Pt nanoflowers possess excellent catalytic activity and large electroactive surface area, contributing to enzymatic activity preservation and long-term stability. This work provides a promising platform for long-time monitoring of other neurotransmitters and secretions in cellular glycolysis and apoptosis processes in the future.

摘要

三磷酸腺苷(ATP)在能量代谢和细胞外嘌呤能信号转导中起着至关重要的作用。本文首次报道了一种通过简便有效的无模板电沉积制备的 3D 双金属 Au/Pt 纳米花修饰的 ATP 微电极生物传感器,实现了对细胞 ATP 分泌的局部检测。该 ATP 生物传感器通过共固定葡萄糖氧化酶和己糖激酶来构建,具有长期稳定性(在 4°C 下放置 14 天后仍保持初始值的 79.39±9.15%)和高选择性,检测限低至 2.5 μM(S/N=3)。然后,将所得的 ATP 生物传感器用于通过高 K 溶液刺激对来自活细胞(PC12)分泌的 ATP 进行直接原位监测。获得的电流约为 21.6±3.4 nA(N=6),对应于从细胞释放的 12.2±2.8 μM ATP,正好在微摩尔范围内,与建议水平一致。3D 双金属 Au/Pt 纳米花具有出色的催化活性和较大的电化学活性表面积,有助于保持酶的活性和长期稳定性。这项工作为未来在细胞糖酵解和细胞凋亡过程中对其他神经递质和分泌物进行长时间监测提供了一个有前途的平台。

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