Ultrasensitive Electrochemical Immunoassays of IgG and CA125 Based on Glucose Oxidase-Catalyzed Signal Amplification with Gold Staining.

Herein, we propose an ultrasensitive electrochemical immunosensor based on glucose oxidase labeling and enzyme-catalyzed Au staining. In brief, the primary antibody (Ab1), bovine serum albumin, an antigen and then a bionanocomposite that contains a second antibody (Ab2), poly(3-anilineboronic acid) (PABA), Au nanoparticles (AuNPs) and glucose oxidase (GOx) are modified on a glassy carbon electrode coated with multiwalled carbon nanotubes, yielding a corresponding sandwich-type immunoelectrode. In the presence of glucose, a chemical reduction of NaAuCl4 by enzymatically generated H2O2 can precipitate a lot of gold on the Ab2-PABA-AuNPs-GOx immobilized immunoelectrode. In situ anodic stripping voltammetry (ASV) detection of gold in 8 μL 1.0 M aqueous HBr-Br2 is conducted for the antigen assay, and the ASV detection process takes approximately 6 min. This method is employed for the assay of human immunoglobulin G (IgG) and human carbohydrate antigen 125 (CA125), which demonstrates exceptional sensitivity, high selectivity and fewer required reagents/samples. The achieved limits of detection (S/N = 3) by the method are 0.25 fg mL-1 for IgG (approximately equivalent to containing 1 IgG molecule in the 1 microlitre of the analytical solution) and 0.1 nU mL-1 for CA125, which outperforms many previously reported results.