基于 H2O2 敏感纳米簇的荧光生物传感器用于乙酰胆碱的检测。
A fluorometric biosensor based on H2O2-sensitive nanoclusters for the detection of acetylcholine.
机构信息
Key Laboratory of Chemical Biology & Traditional Chinese Medicine Research, Ministry of Education, Hunan Normal University, Changsha 410081, China.
Key Laboratory of Chemical Biology & Traditional Chinese Medicine Research, Ministry of Education, Hunan Normal University, Changsha 410081, China.
出版信息
Biosens Bioelectron. 2014 Sep 15;59:289-92. doi: 10.1016/j.bios.2014.03.054. Epub 2014 Apr 2.
We have constructed a simple, cost-effective, label-free, selective and sensitive biosensor based on the fluorescence quenching of protein-stabilized nanoclusters (NCs) for the detection of acetylcholine (ACh). In this assay, acetylcholineaterase (AChE) catalyzes the hydrolysis of ACh to form choline, which in turn is oxidized by choline oxidase (ChOx) to generate H2O2 which induces the fluorescence quenching of NCs. By virtue of this specific response, the detection limit of a fluorescent biosensor for determination of ACh was 5 pM (signal/noise=3), and the linear range was 0.1-20 nM. In addition, we use the proposed method to detect ACh in human blood, in which the concentration of ACh is ca. 6.83 nM, and the recoveries of the spiked sample range from 93% to 102%. The result agree with a commercial assay kit, suggesting that the excellent biosensor can be used in the practice detection of ACh.
我们构建了一种简单、经济、无标记、选择性和灵敏的基于蛋白质稳定纳米团簇（NCs）的荧光猝灭的生物传感器，用于检测乙酰胆碱（ACh）。在该测定中，乙酰胆碱酯酶（AChE）催化 ACh 的水解形成胆碱，胆碱继而被胆碱氧化酶（ChOx）氧化生成 H2O2，诱导 NCs 的荧光猝灭。凭借这种特异性反应，荧光生物传感器测定 ACh 的检测限为 5 pM（信号/噪声=3），线性范围为 0.1-20 nM。此外，我们使用所提出的方法检测人血液中的 ACh，其中 ACh 的浓度约为 6.83 nM，加标样品的回收率范围为 93%至 102%。结果与商业测定试剂盒一致，表明该出色的生物传感器可用于 ACh 的实际检测。
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