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

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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 H2O2, 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.

Graphical abstract

Nanozyme based on Prussian blue nanocube (PB NCs) loaded with copper nanoparticles was synthesized and utilized for detecting the activity of alkaline phosphatase (ALP).

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Funding

The authors thank the Hunan Provincial Science and Technology Plan Project, China (No. 2019TP1001) and Innovation-Driven Project of Central South University (2020CX002) for the financial support of this work.

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Authors and Affiliations

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

    Shengnan Fan, Xingxing Jiang & Minghui Yang

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

    Xianggui Wang

  3. Hunan Key Laboratory of Ophthalmology, Changsha, 410078, Hunan, China

    Xianggui Wang

Authors
  1. Shengnan Fan
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  2. Xingxing Jiang
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  3. Minghui Yang
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  4. Xianggui Wang
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Corresponding authors

Correspondence to Minghui Yang or Xianggui Wang.

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All experiments were in accordance with the guidelines of the National Institute of Health, China, and approved by the Institutional Ethical Committee (IEC) of the Second Xiangya Hospital that attached to Central South University.

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Fan, S., Jiang, X., Yang, M. et al. Sensitive colorimetric assay for the determination of alkaline phosphatase activity utilizing nanozyme based on copper nanoparticle-modified Prussian blue. Anal Bioanal Chem 413, 3955–3963 (2021). https://doi.org/10.1007/s00216-021-03347-y

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  • DOI: https://doi.org/10.1007/s00216-021-03347-y

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