An impedance array biosensor for detection of multiple antibody–antigen interactions

Electrochemical impedance spectroscopy (EIS) combined with a gold electrode array was developed to detect multiple antibody–antigen interactions. Hepatitis B surface antigen (HBsAg), as a model sample, was employed to evaluate the characteristics of the biosensor. The array was fabricated by immobilizing antibodies on the self-assembled molecules surface of the electrodes. The surface characteristics of the array during the binding process including the antibody–antigen conjugation and the sandwich complex with HRP-labeled antibody, as well as the precipitation layer, were characterized by atomic force microscopy (AFM) and electrochemical impedance spectroscopy, respectively. A linear relationship between electron-transfer resistance and the concentrations of HBsAg ranged from 10 pg ml−1 to 1 ng ml−1 and the detection limit of 10 pg ml−1 was obtained. 100 pg ml−1 antigen samples, such as rat IgG, HBsAg and HBeAg, as well as the antigen mixture, were incubated with the relative antibody-modified electrodes on the array. No obvious cross-talk reaction was observed. All these results confirm the feasibility of applying electrochemical impedance spectroscopy to the electrode array.