College of Food Science and Engineering, Jilin University, Changchun 130025, China.
College of New Energy and Environment, Jilin University, Changchun 130012, China.
J Mater Chem B. 2019 Nov 14;7(42):6508-6514. doi: 10.1039/c9tb01390c. Epub 2019 Oct 2.
In this work, a fluorometric and colorimetric analysis of alkaline phosphatase (ALP) activity was developed based on nanozymes. The nanozymes were composed of nucleotides (ATP, ADP and AMP) coordinated with copper ions. All three kinds of nanozymes (ATP-Cu, ADP-Cu and AMP-Cu) exhibited polyphenol oxidase (PPO)-mimic activity by catalyzing a chromogenic reaction of 2,4-dichlorophenol (2,4-DP) and 4-aminoantipyrine (4-AP). However, there were obvious differences in the PPO-like activity and the fluorescence of the three nanozymes produced from the same concentration of nucleotides (keeping the concentration of Cu unchanged at 5 mM). The catalytic activities of produced ADP-Cu and AMP-Cu were obviously higher than that of ATP-Cu at a certain nucleotide concentration of 3 mM. In addition, when ATP was hydrolyzed into ADP and AMP by ALP, more nanozymes were produced and the catalytic activity of the system was enhanced, which resulted in an obvious increase of the colorimetric signal. The signal intensity was proportional to ALP concentration in the range of 0-30 U L, and the detection limit for ALP was 0.3 U L from the colorimetric detection. Moreover, the fluorescence intensity of the produced nanozymes was also proportional to the ALP concentration in the range of 1-30 U L and the detection limit was 0.45 U L from the fluorescence detection. A fluorometric and colorimetric sensing ALP method was thus established. The method showed a high selectivity for ALP activity compared with proteins, amino acids and other interference components. Furthermore, the proposed method was also used to detect ALP activity in human serum samples, which showed great potential for diagnostic and practical purposes.
在这项工作中，基于纳米酶发展了一种碱性磷酸酶（ALP）活性的荧光和比色分析。纳米酶由核苷酸（ATP、ADP 和 AMP）与铜离子配位组成。三种纳米酶（ATP-Cu、ADP-Cu 和 AMP-Cu）均通过催化显色反应 2,4-二氯苯酚（2,4-DP）和 4-氨基安替比林（4-AP）表现出多酚氧化酶（PPO）模拟活性。然而，从相同浓度的核苷酸（保持 Cu 浓度不变为 5 mM）产生的三种纳米酶的 PPO 样活性和荧光存在明显差异。在一定的核苷酸浓度 3 mM 下，ADP-Cu 和 AMP-Cu 产生的催化活性明显高于 ATP-Cu。此外，当 ALP 将 ATP 水解为 ADP 和 AMP 时，会产生更多的纳米酶，从而增强了系统的催化活性，导致比色信号明显增加。在 0-30 U L 范围内，信号强度与 ALP 浓度成正比，比色检测的 ALP 检测限为 0.3 U L。此外，所产生的纳米酶的荧光强度也与 ALP 浓度在 1-30 U L 范围内成正比，荧光检测的 ALP 检测限为 0.45 U L。因此建立了一种荧光和比色检测 ALP 的方法。与蛋白质、氨基酸和其他干扰成分相比，该方法对 ALP 活性具有很高的选择性。此外，该方法还用于检测人血清样品中的 ALP 活性，具有很大的诊断和实际应用潜力。
