Determination of alkaline phosphatase activity based on enzyme-triggered generation of a thiol and the fluorescence quenching of silver nanoclusters.
Mikrochim Acta
; 186(3): 180, 2019 02 15.
Article
em En
| MEDLINE
| ID: mdl-30771096
ABSTRACT
A fluorimetric method is described for the determination of alkaline phosphatase (ALP) activity. It is based on the use of polyethyleneimine-coated silver nanoclusters (AgNCs), which display an intense blue fluorescence peaking at 450 nm (under 375 nm excitation). ALP catalyzes the dephosphorylation of the thiophosphate amifostine to generate a thiol that binds to the AgNCs and causes its fluorescence to be quenched. Under the optimal experimental conditions, fluorescence linearly drops in the 0.08-2.0 U L-1 ALP activity range, and the limit of detection is 0.02 U L-1. The method was successfully applied to the determination of ALP activity in spiked human serum samples. Graphical abstract Alkaline phosphatase (ALP) catalyzes the degradation of amifostine with a generation a thiol product. The thiol quenches the fluorescence of silver nanoclusters, and a method for the detection of ALP down to 0.02 U L-1 was developed.
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Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Prata
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Fosfatase Alcalina
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Nanopartículas Metálicas
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Ensaios Enzimáticos
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Fluorometria
Limite:
Humans
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article