State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences , Changchun, Jilin 130022, China.
University of Science and Technology of China , Hefei, Anhui 230026, China.
ACS Appl Mater Interfaces. 2018 Feb 21;10(7):6560-6569. doi: 10.1021/acsami.7b18816. Epub 2018 Feb 9.
As an essential and universal hydrolase, alkaline phosphatase (ALP) has been identified as a crucial indicator of various diseases. Herein, we, for the first time, expanded the application of fluorescent polydopamine (F-PDA) nanoparticles to nanoquencher-based biosensing system, as well as discovered the reversible quenching effect of manganese dioxide (MnO) nanosheets on the fluorescence of F-PDA nanoparticles and intensively confirmed the quenching mechanism of Förster resonance energy transfer by using transmission electron microscopy, UV-vis, Fourier transform infrared spectroscopy, and fluorescence lifetime experiments. By means of the ALP-triggered generation of ascorbic acid (AA) from the substrate ascorbic acid 2-phosphate, the AA-triggered reduction of MnO nanosheets to Mn, as well as the clear quenching mechanism of F-PDA nanoparticles by MnO nanosheets, we have developed a label-free, low-cost, visual, and facile synthetic fluorescent biosensor for convenient assay of ALP activity. The fluorescent bioassay shows a good linear relationship from 1 to 80 mU/mL (R = 0.999), with a low detection limit of 0.34 mU/mL, and the excellent applicability in human serum samples demonstrates potential applications in clinical diagnosis and biomedical research.
作为一种必需且普遍的水解酶,碱性磷酸酶(ALP)已被确定为各种疾病的重要指标。在此,我们首次将荧光聚多巴胺(F-PDA)纳米粒子的应用扩展到基于荧光猝灭剂的生物传感系统,并发现了二氧化锰(MnO)纳米片对 F-PDA 纳米粒子荧光的可逆猝灭效应,并通过透射电子显微镜、紫外-可见吸收光谱、傅里叶变换红外光谱和荧光寿命实验,深入证实了Förster 共振能量转移的猝灭机制。通过 ALP 从作为基质的抗坏血酸 2-磷酸中引发抗坏血酸(AA)的生成,AA 引发 MnO 纳米片还原为 Mn,以及 MnO 纳米片对 F-PDA 纳米粒子的明显猝灭机制,我们开发了一种无标记、低成本、可视化且简便的合成荧光生物传感器,用于方便测定 ALP 活性。荧光生物测定法显示出 1 至 80 mU/mL(R = 0.999)的良好线性关系,检测限低至 0.34 mU/mL,在人血清样本中的优异适用性证明了其在临床诊断和生物医学研究中的潜在应用。