Ratiometric electrochemiluminescent strategy regulated by electrocatalysis of palladium nanocluster for immunosensing

This work designed a novel ratiometric electrochemiluminescence (ECL) immunosensing approach based on two different ECL emitters: CdS quantum dots (QDs) as cathodic emitter and luminol as anodic emitter. The ECL immunosensor was constructed by a layer-by-layer modification of CdS QDs, Au nanoparticles and capture antibody on a glassy carbon electrode. With hydrogen peroxide as ECL coreactant, the immunosensor showed a cathodic ECL emission of CdS QDs at −1.5 V (vs Ag/AgCl) in air-saturated pH 8.0 buffer. Upon the formation of sandwich immunoassay, the lumiol/palladium nanoclusters (Pd NCs)@graphene oxide probe was introduced to the electrode. Therefore, the cathodic ECL intensity decreased and luminol anodic ECL emission was appeared at +0.3 V (vs Ag/AgCl) owing to the competition of the coreactant of hydrogen peroxide. Using carcino-embryonic antigen as model, this ratiometric ECL strategy could be used for immunoassay with a linear range of 1.0–100 pg mL−1 and a detection limit of 0.62 pg mL−1. The enhanced ratiometric ECL signal resulted from the high density and excellent electrocatalysis of the loaded Pd NCs. The immunosensor exhibited good stability and acceptable fabrication reproducibility and accuracy, showing a great promising for clinical application. This electrocatalysis-regulated ratiometric ECL provides a new concept for ECL measurement, and could be conveniently extended for detection of other protein biomarkers.