Alkaline phosphatase (ALP), an important hydrolase involved in dephosphorylation, is a common clinical indicator of many diseases. In the present study, we constructed a novel electrochemical sensor using amifostine as the substrate of ALP and activators regenerated by electron transfer atom transfer radical polymerization (ARGET ATRP) as a signal amplification strategy for sensitive determination of ALP activity. In particular, in the presence of ALP, the phosphate group of amifostine was hydrolyzed to form a sulfhydryl group, which could attach to a gold electrode via a sulfur-gold bond. Then, the initiator α-bromophenylacetic acid (BPAA) was linked to the hydrolysis product of amifostine through an amide bond, resulting in the production of electroactive polymer chains on the gold electrode by the monomer ferrocenylmethyl methacrylate (FMMA) via ARGET ATRP. Under optimal parameters, the electrochemical sensor demonstrated a limit of detection (LOD) of 1.71 mU mL-1 with a linear range of 5-100 mU mL-1. In addition to satisfactory selectivity, the potential application of this approach for ALP activity detection in human serum samples was demonstrated. Due to its efficiency, simplicity of operation, and cost-effectiveness, the proposed electrochemical sensor has great promise as a universal method for ALP assays and inhibitor screening.
Keywords: ATRP; Alkaline phosphatase; Amifostine; Biomarker; Electrochemical sensor.
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