Pharmacy College, Henan University of Chinese Medicine, Zhengzhou, 450046, People's Republic of China.
School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing, 210094, People's Republic of China.
Mikrochim Acta. 2022 Feb 7;189(3):84. doi: 10.1007/s00604-022-05189-6.
A novel fluorescence assay is proposed through activators regenerated by electron transfer atom transfer radical polymerization (ARGET ATRP) strategy for alkaline phosphatase (ALP) activity detection. First of all, 2-bromo-2-methylpropionic acid (BMP) was employed as the initiator to modify on the surface of the magnetic nanoparticle (FeO-MNP) by amide bonding. Then, ascorbic acid (AA) produced by ALP catalyzed the phosphate group removal from L-ascorbic acid 2-phosphate sesquimagnesium salt hydrate (AAPS), which underwent a redox reaction with Cu(II) and the product Cu(I) triggered the ARGET ATRP reaction. Finally, a strong fluorescent signal could be detected at 514 nm due to numerous fluorescent monomers being grafted to the FeO-MNPs surface (E = 490 nm, E = 514 nm). Under optimal experimental conditions, the linear range of this fluorometric assay for ALP activity was 1-80 mU mL, and the detection limit was 0.68 mU mL. The method exhibited excellent selectivity and satisfactory results were obtained in the inhibition rate and human serum experiments. Therefore, this ALP activity detection strategy has great potential for clinically relevant disease detection and drug screening. A novel fluorescence strategy for alkaline phosphatase activity detection based on the dephosphorylation property of alkaline phosphatase and ARGET ATRP reaction.
通过电子转移原子转移自由基聚合（ARGET ATRP）策略提出了一种新颖的荧光分析方法，用于碱性磷酸酶（ALP）活性检测。首先，将 2-溴-2-甲基丙酸（BMP）用作引发剂，通过酰胺键修饰在磁性纳米粒子（FeO-MNP）表面上。然后，ALP 催化 L-抗坏血酸 2-磷酸半镁盐水合物（AAPS）中的磷酸基团去除，生成抗坏血酸（AA），该物质与 Cu(II) 发生氧化还原反应，产物 Cu(I) 引发 ARGET ATRP 反应。最后，由于大量荧光单体接枝到 FeO-MNPs 表面上，在 514nm 处可以检测到强荧光信号（E=490nm，E=514nm）。在最佳实验条件下，该荧光分析法用于 ALP 活性的线性范围为 1-80mU mL，检测限为 0.68mU mL。该方法表现出优异的选择性，在抑制率和人血清实验中均取得了令人满意的结果。因此，这种基于碱性磷酸酶的去磷酸化性质和 ARGET ATRP 反应的碱性磷酸酶活性检测策略具有很大的临床相关疾病检测和药物筛选潜力。
