Sandwich-type electrochemical immunosensor for carcinoembryonic antigen detection based on the cooperation of a gold–vertical graphene electrode and gold@silica–methylene blue

In this study, a sandwich-type electrochemical (EC) immunosensor was proposed to detect a carcinoembryonic antigen (CEA) based on Au-graphene and Au@SiO2-methylene blue (MB). The Au nanoparticles (NPs)-vertical graphene (VG) electrode efficiently amplifies the response signal by immobilizing a large amount of the coating antibody (Ab) and is characterized by excellent electrocatalytic activity. The MB nanodot-loaded Au@SiO2 carriers with core-shell nanostructure and detection Ab were used to construct the Ab-Au@SiO2-MB label, which improved the sensitivity due to the high EC signal of MB nanodots and the high labeling effect between the detection Ab and MB probe. A novel double-Ab sandwich strategy was developed to further improve the sensitivity and stability based on the same specificity of the coating and detection Abs for the recognition of CEA. Under optimal conditions, the developed EC sensor exhibited a wide linear range from 1 fg mL-1 to 100 ng mL-1, with an ultralow detection limit of 0.8 fg mL-1 (S/N = 3). The feasibility in the clinical application of the EC sensor was verified by the in vitro detection of CEA in human serum.