Electrochemical immunosensors for the simultaneous detection of two tumor markers

The microfabrication of electrochemical immunosensors for the simultaneous detection of two protein analytes is described. The sensors consisted of two iridium oxide electrodes (1-mm diameter) patterned on a glass substrate. Capture antibodies were immobilized on the porous iridium oxide electrodes by covalent attachment using (3-aminopropyl)triethoxysilane and glutaraldehyde. The spatial separation of the electrodes (2.5 mm) enabled simultaneous electrochemical immunoassays to be conducted without cross-talk between the electrodes. Proteins were measured using electrochemical ELISA, and detection was achieved by electrochemically oxidizing alkaline phosphatase-generated hydroquinone. Sensors for the simultaneous detection of goat IgG and mouse IgG, and for the tumor markers CEA and AFP, were developed. The sensors had detection limits of 1, 2, 1.2, and 1 ng/mL for goat IgG, mouse IgG, CEA, and AFP, respectively.