Grafting of polymers via ring-opening polymerization for electrochemical assay of alkaline phosphatase activity

Abstract

As an important hydrolytic enzyme, abnormal activity of alkaline phosphatase (ALP) is closely associated with a variety of diseases. It has been identified as an important diagnostic indicator for clinical hepatobiliary and bone diseases. Herein, a novel electrochemical sensor based on signal amplification strategy through ring-opening polymerization (ROP) has been developed to assay of ALP activity. First of all, 3-mercaptopropanoic acid (MPA) was employed as a cross-linking agent to attach O-phosphoethanolamine to the electrode surface via amide bond. Then, ALP catalyzed the hydrolysis of phosphate monoester structures to hydroxyl groups, which could initiate ROP reaction. The polymer grafted on the electrode surface contains a large number of ferrocene electroactive molecules, which effectively increased the signal output of the electrochemical sensor and improved the sensitivity of ALP activity detection. Under optimum conditions, this electrochemical sensor rendered a satisfactory linear dependence over the range from 20 to 120 mU mL−1, with a low detection limit of 0.66 mU mL−1. Furthermore, this strategy presented satisfactory selectivity and interference resistance in human serum sample, and compared with clinical data, the relative error of the results obtained by this method was less than 5%. Thus, this method showed considerable potential for the detection of ALP activity in clinical application.