Electrochemical detection of alkaline phosphatase activity via atom transfer radical polymerization

Alkaline phosphatase (ALP) activity is a diagnostic indicator for a variety of clinical diseases. In this study, an electrochemical method for detecting ALP activity through activators regenerated by electron transfer atom transfer radical polymerization (ARGET ATRP) was developed. Specifically, 3-mercaptopropionic (MPA) was firstly fixed on the electrode through sulfur-gold bonding. Subsequently, α-bromophenylacetic acid (BPAA) as initiator was attached to MPA through the recognized carboxylate-Zr⁴⁺-phosphate chemistry. Finally, in the existence of ALP, L-Ascorbic acid 2-phosphate sesquimagnesium salt hydrate (AAPS) was hydrolyzed to produce ascorbic acid (AA) which participated in the ARGET ATRP reaction, grafting polymer containing plenty of ferrocene electroactive probes on the surface of electrode. Under optimal experimental conditions, this method had a linear scope of 20-200 mU mL^-1, and a limit of detection (LOD) of 1.64 mU mL^-1. In addition, the proposed method had good selectivity as well as anti-interference capability, with satisfactory results in inhibition rate and human serum experiments. By merits of good analytical performance, easy operation, and low cost, such a method for ALP activity detection has promising applications in ALP-related disease detection and inhibitor screening.