利用基于铜纳米粒子修饰的普鲁士蓝的纳米酶的灵敏比色法测定碱性磷酸酶活性。

Sensitive colorimetric assay for the determination of alkaline phosphatase activity utilizing nanozyme based on copper nanoparticle-modified Prussian blue.

机构信息

Hunan Provincial Key Laboratory of Micro & Nano Materials Interface Science, College of Chemistry and Chemical Engineering, Central South University, Changsha, 410083, Hunan, China.

Eye Center of Xiangya Hospital, Central South University, Changsha, 410011, Hunan, China.

出版信息

Anal Bioanal Chem. 2021 Jun;413(15):3955-3963. doi: 10.1007/s00216-021-03347-y. Epub 2021 Apr 22.

DOI:10.1007/s00216-021-03347-y

PMID:33885935

Abstract

Nanozyme based on Prussian blue nanocubes (PB NCs) loaded with copper nanoparticles (Cu@PB NCs) was synthesized. The peroxidase (POD)-like activity of Cu@PB NCs was studied and utilized for detecting the activity of alkaline phosphatase (ALP). The Cu@PB NCs possess higher POD-like activity compared with PB NCs and natural horseradish peroxidase (HRP) due to the loading of copper nanoparticles. 3,3',5,5'-Tetramethylbenzidine (TMB) can be oxidized to oxTMB in the presence of Cu@PB NCs and HO, generating blue-colored compound, while introduction of pyrophosphate (PPi) leads to the POD-like activity of Cu@PB NCs decreased obviously. In the presence of ALP, PPi was hydrolyzed and then the POD-like activity of Cu@PB NCs was restored. So, according to the change of the POD-like activity of Cu@PB NCs, a sensitive colorimetric assay for ALP activity was reported. The limit of detection of the assay is 0.08 mU/mL, with linear range from 0.1 to 50 mU/mL. In addition, the assay was also applied for screening the inhibitors of ALP. Nanozyme based on Prussian blue nanocube (PB NCs) loaded with copper nanoparticles was synthesized and utilized for detecting the activity of alkaline phosphatase (ALP).

摘要

基于负载铜纳米粒子的普鲁士蓝纳米立方（PB NCs）的纳米酶被合成。研究了 Cu@PB NCs 的过氧化物酶（POD）样活性，并将其用于检测碱性磷酸酶（ALP）的活性。由于负载了铜纳米粒子，Cu@PB NCs 具有比 PB NCs 和天然辣根过氧化物酶（HRP）更高的 POD 样活性。在 Cu@PB NCs 和 HO 的存在下，3,3',5,5'-四甲基联苯胺（TMB）可以被氧化为 oxTMB，生成蓝色化合物，而焦磷酸（PPi）的引入导致 Cu@PB NCs 的 POD 样活性明显降低。在 ALP 的存在下，PPi 被水解，然后 Cu@PB NCs 的 POD 样活性得到恢复。因此，根据 Cu@PB NCs 的 POD 样活性的变化，报道了一种用于检测 ALP 活性的灵敏比色法。该测定的检测限为 0.08 mU/mL，线性范围为 0.1 至 50 mU/mL。此外，该测定还用于筛选 ALP 的抑制剂。