A new immunosensing method by galactoseoxidase-mediated electrocatalysis using a virtualbeaker array

We report a novel and convenient method for the determination of glycoproteins, especially antibodies, using galactose oxidase (GAO) on the basis of the contents of galactosyl and N-acetylgalactosaminyl residues in carbohydrate chains of glycoproteins. GAO converts galactose residues to their corresponding aldehyde and H₂O₂, the latter being electroactive and quantifiable by DC amperometry. The method does not require processes such as antibody labeling or the use of enzyme-tagged secondary antibodies. For an array-type immunosensing, the platform surface for antigen immobilization was specially designed by using differentiated surface wetting property of hydrophobic and hydrophilic patterns. We patterned the hydrophobic surface of the poly(dimethylsiloxane) substrate by microcontact printing with the poly(amidoamine) dendrimer ink, providing hydrophilic patterns on a hydrophobic base substrate. By applying aqueous solution on the patterned surface, an array of free-standing water droplets was made. With the prepared virtual beaker array, electrochemical immunosensing was performed by using anti-dinitrophenyl-IgG as a model target protein. From immunoassay with GAO-mediated electrocatalysis, a good correlation in amperometric signal with the target IgG was registered. The total assay time was about 20 min, including antibody recognition and signal registration.