A colorimetric alkaline phosphatase biosensor based on p-aminophenol-mediated growth of silver nanoparticles.

Alkaline phosphatase (ALP) is an enzyme that catalyzes the dephosphorylation of proteins, nucleic acids, and biomolecules. It is a potential biomarker for diverse diseases such as breast cancer, osteopenia, and hepatobiliary. Herein, we developed a colorimetric sensor for the ALP assay based on its enzymatic activity to dephosphorylate the p-aminophenol phosphate (pAPP) into pAP. In a solution containing silver nanoparticles (AgNPs) and Ag ions prepared using a low concentration of NaBH, pAP mediates the growth of AgNPs by reducing the concentration of Ag ions to enhance the intensity of localized surface plasmon resonance as the pAPP cannot induce a reduction of the remaining Ag due to the masking of the hydroxyl with phosphate. The quantitative assay of the ALP was demonstrated via the colorimetric detection of the pAP-mediated growth of AgNPs in the presence of an ALP. The highly sensitive enzymatic growth of AgNPs provided a wider dynamic linear range of 0.5-225 U/L with a lower limit of detection of 0.24 U/L than that previously reported. The use of pAP resulted in excellent selectivity of the sensor for the ALP assay in human serum, yielding a high recovery rate and a high precision of 99.2 ± 1.5 % for the standard addition method.