Fluorescent assay of alkaline phosphatase activity via atom transfer radical polymerization.

Abstract

A novel fluorescence assay is proposedthrough activators regenerated by electron transfer atom transfer radical polymerization (ARGET ATRP) strategy for alkaline phosphatase (ALP) activity detection. First of all, 2-bromo-2-methylpropionic acid (BMP) was employed as the initiator to modify on the surface of the magnetic nanoparticle (Fe3O4-MNP) by amide bonding. Then, ascorbic acid (AA) produced by ALP catalyzed the phosphate group removal from L-ascorbic acid 2-phosphate sesquimagnesium salt hydrate (AAPS), which underwent a redox reaction with Cu(II) and the product Cu(I) triggered the ARGET ATRP reaction. Finally, a strong fluorescent signal could be detected at 514nm due to numerous fluorescent monomers being grafted to the Fe3O4-MNPs surface (Ex = 490nm, Em = 514nm). Under optimal experimental conditions, the linear range of this fluorometric assay for ALP activity was 1-80mUmL-1, and the detection limit was 0.68mUmL-1. Themethod exhibited excellent selectivity and satisfactory results were obtained in the inhibition rate and human serum experiments. Therefore, this ALP activity detection strategy has great potential for clinically relevant disease detection and drug screening. A novel fluorescence strategy for alkaline phosphatase activity detection based on the dephosphorylation property of alkaline phosphatase and ARGET ATRP reaction.