Sandwich-type electrochemical immunosensor for sensitive detection of CEA based on the enhanced effects of Ag NPs@CS...

An ultrasensitive sandwich-type electrochemical immunosensor was designed by using gold nanoparticles (Au NPs) as the substrate material and microporous carbon spheres (CS) loading silver nanoparticles (Ag NPs) spaced Hemin/reduced graphene oxide (Hemin/rGO) porous composite materials (Ag NPs@CS-Hemin/rGO) as the detection antibodies (Ab₂) label for detecting carcinoembryonic antigen (CEA). The Au NPs with good electrical conductivity and biocompatibility could accelerate the electron transfer on the electrode interface and enhance the load capacity of capture antibodies (Ab₁). Hemin is peroxidase-like substance which has excellent catalytic ability for H₂O₂ reduction but easy to molecular aggregation and oxidative self-destruction. Reduced graphene oxide (rGO) is a good supporting material for Hemin to mitigate this disadvantage. CS loading Ag NPs (Ag NPs@CS) as the spacer inserts into Hemin/rGO sheet can overcome the irreversible stacking of rGO, and form complex porous structure which exposes more active sites of Hemin. Moreover, Ag NPs loaded on CS also has catalytic ability for H₂O₂ reduction. Thus the Ag NPs@CS-Hemin/rGO used as the Ab₂ label has a large working surface area and high utilization rate, which heightens the catalytic ability for H₂O₂ reduction to amplify the current signal effectually. The current signal and the logarithm of CEA concentration presented a wide linear response range of 20 fg/mL to 200 ng/mL, and the detection limit of CEA was 6.7 fg/mL. Furthermore, the designed immunosensor exhibited a good reproducibility, selectivity and stability, which confirms a broad development prospect when applying it in early clinical detection.