Electrochemical Immunoassay for Detecting Hippuric Acid Based on the Interaction of Osmium-Antigen Conjugate Films with Antibody on Screen Printed Carbon Electrodes

摘要

An electrochemical immunoassay based on osmium-hippuric acid (HA) conjugate films onto the electrode is presented for the detection of urinary HA. This is the first report on the use of the oxidative electropolymerization of 5-amino-1,10-phenanthroline (5-NH2-phen) for immobilizing an antigen, osmium-conjugated HA. As a redox mediator, [Os(5-amino-1,10-phenanthroline)2(4-aminomethylpyridine-HA)Cl]+/2+ (Os-phen-HA) was successfully synthesized and electropolymerized onto the screen-printed carbon electrodes (SPCEs). The interaction between osmium-HA conjugate films and antibody-HA (anti-HA) was performed by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The electrical signals were linearly proportional to urinary HA in the range of 0.1-5.0 mg/mL, which is sufficient for use as an immunosensor using a cutoff concentration of 2.0 mg/mL in urine samples. The proposed electrochemical immunoassay method can be extended to various applications for detecting a wide range of different small antigens in the health care area.