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 NaBH₄, 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.