基于 DNA 模板铜/银纳米簇的乙酰胆碱酯酶活性和抑制剂检测的荧光分析法。
A fluorometric assay for acetylcholinesterase activity and inhibitor detection based on DNA-templated copper/silver nanoclusters.
机构信息
State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.
出版信息
Biosens Bioelectron. 2013 Sep 15;47:345-9. doi: 10.1016/j.bios.2013.03.038. Epub 2013 Mar 27.
A novel label-free, rapid, cost-effective, and highly sensitive fluorometric sensor has been constructed for the detection of acetylcholinesterase (AChE) activity and its inhibitor based on the fluorescence quenching of DNA-templated copper/silver nanoclusters (DNA-Cu/AgNCs). In this assay, AChE catalyzes the hydrolysis of acetylthiocholine (ATCh) to form thiocholine which induces fluorescence quenching of DNA-Cu/AgNCs. The AChE activity could be detected as low as 0.05mU/mL and with a linear range from 0.05 to 2.0mU/mL. This assay offers a very convenient "mix and detect" approach for AChE activity. On the other hand, tacrine and organophosphorus pesticides (OPPs) were employed to inhibit the hydrolysis of ATCh, which could eliminate the fluorescence quenching of DNA-Cu/AgNCs. The IC50 of tacrine and methamidophos were estimated to be 16.9nM and 0.075mg/L, respectively. This method was also used to detect spiked OPPs in agricultural products successfully. The present work may expand the use of DNA-Cu/AgNCs to the field of enzyme sensors.
一种新型的无标记、快速、经济且高灵敏度的荧光传感器已被构建，用于基于 DNA 模板铜/银纳米团簇（DNA-Cu/AgNCs）的荧光猝灭来检测乙酰胆碱酯酶（AChE）活性及其抑制剂。在该测定中，AChE 催化乙酰硫代胆碱（ATCh）的水解，形成硫代胆碱，从而导致 DNA-Cu/AgNCs 的荧光猝灭。AChE 活性可低至 0.05mU/mL 检测，线性范围为 0.05 至 2.0mU/mL。该测定提供了一种非常方便的“混合检测”方法，用于检测 AChE 活性。另一方面，他克林和有机磷农药（OPPs）被用来抑制 ATCh 的水解，这可以消除 DNA-Cu/AgNCs 的荧光猝灭。他克林和甲胺磷的 IC50 分别估计为 16.9nM 和 0.075mg/L。该方法还成功地用于检测农产品中添加的 OPPs。本工作可能会将 DNA-Cu/AgNCs 的应用扩展到酶传感器领域。
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