Fluorometric determination of the activity of alkaline phosphatase based on the competitive binding of gold nanoparticles and pyrophosphate to CePO4:Tb nanorods
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A fluorometric method is described for the determination of the activity of alkaline phosphatase (ALP). It relies on the competition between gold nanoparticles (AuNPs) and pyrophosphate (PPi) for the coordination sites on the surface of CePO4:Tb nanorods. The green fluorescence of the CePO4:Tb is reduced in the presence of AuNPs due to fluorescence resonance energy transfer (FRET), but can be restored on addition of PPi due to the stronger affinity of PPi to the CePO4:Tb. In the presence of ALP, PPi is hydrolyzed to form phosphate which has much weaker affinity for the CePO4:Tb. Hence, the AuNPs will reassemble on the CePO4:Tb, and fluorescence is reduced. Fluorescence drops linearly in the 0.2 to 100 U·L−1 activity range, and the detection limit is 60 mU·L−1 (at S/N = 3). The method does not require any modification of the surface of the CePO4:Tb and is highly sensitive and selective. The inhibition of ALP activity by Na3VO4 was also studied. In our perception, the method may find application in the diagnosis of ALP-related diseases, in screening for inhibitors, and in studies on ALP-related functions in biological systems.
KeywordsFluorescence resonance energy transfer Fluorescence quenching Dephosphorylation Phosphate Rare-earth nanoparticle Affinity Alkaline phosphatase inhibitor
This work is supported by the National Natural Science Foundation of China (21775065, 21675078).
Compliance with Ethical Standards
The author(s) declare that they have no competing interests.
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