A ratiometric fluorescence probe based on carbon dots for discriminative and highly sensitive detection of acetylcholinesterase and butyrylcholinesterase in human whole blood
A ratiometric fluorescence probe based on carbon dots for discriminative and highly sensitive detection of acetylcholinesterase and butyrylcholinesterase in human whole blood
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基于碳点的比例荧光探针，用于人全血中乙酰胆碱酯酶和丁酰胆碱酯酶的判别性和高灵敏度检测
DOI：
10.1016/j.bios.2019.02.031
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发表时间：
2019
影响因子：
12.6
通讯作者：
Hu Qin
中科院分区：
文献类型：
--
作者：
Xu Xiaoman;Cen Yao;Xu Guanhong;Wei Fangdi;Shi Menglan;Hu Qin
研制了一种基于碳点(CDS)的比率荧光探针，用于区分和高灵敏地检测人全血乙酰胆碱酯酶(AChE)和丁酰胆碱酯酶(BChE)活性。当邻苯二胺被Cu2+氧化时，其产物2，3-二氨基吩嗪(OxOPD)由于内滤波作用可有效猝灭硫化镉在460 nm处的荧光，并在570 nm处产生新的发射峰。AChE或BChE催化乙酰硫胆碱或丁基硫代胆碱的水解反应生成硫代胆碱，硫代胆碱的巯基强烈捕获Cu2+，抑制OPD的氧化，从而有效地保持了硫化镉的天然荧光发射。由此得到的荧光强度比作为该探针的信号输出，用于胆碱酯酶(CHS)活性检测。乙酰胆碱酯酶和胆碱酯酶活力分别在0.20~14.0U L−1和0.1U~5.0U L−1之间变化，检出限分别为0.1U L−1和0.04U L−1。此外，他克林和乙丙嗪抑制AChE和BChE的IC50值分别为29.8 nM和132.6 nM。并成功地将该探针用于人全血中AChE和BChE的鉴别测定。这些结果表明，所提出的策略为CHES的临床诊断和药物筛选提供了一个区分、敏感和稳健的分析平台。
A ratiometric fluorescence probe based on carbon dots (CDs) was developed for discriminative and highly sensitive detection of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activity in human whole blood. Wheno-phenylenediamine (OPD) was oxidized by Cu2+, the product 2,3-diaminophenazine (oxOPD) could effectively quench the fluorescence of CDs at 460 nm due to the inner filter effect and gave rise to a new emission peak at 570 nm. The AChE or BChE catalyzed hydrolysis reaction of acetylthiocholine or butyrylthiocholine to generate thiocholine, whose sulfhydryl group strongly captured Cu2+to inhibit the oxidization of OPD, thus effectively preserving the natural fluorescence emission of CDs. The resulting fluorescence intensity ratio served as the signal output of the probe for cholinesterases (ChEs) activity sensing. The activities of AChE and BChE were determined to range from 0.2 to 14.0 U L−1and from 0.1 to 5.0 U L−1, with detection limits of 0.1 U L−1and 0.04 U L−1, respectively. Additionally, the IC50of tacrine and ethopropazine for the inhibition of AChE and BChE were estimated to be 29.8 nM and 132.6 nM, respectively. Moreover, the probe was successfully applied to the discriminative determination of AChE and BChE in human whole blood without any pretreatment. These results suggested that the proposed strategy provided a discriminative, sensitive and robust analytical platform for ChEs clinical diagnostics and drug screening.