Atomically Fe‐anchored MOF‐on‐MOF nanozyme with differential signal amplification for ultrasensitive cathodic electrochemiluminescence immunoassay

The successful application of electrochemiluminescence (ECL) in immunoassays for clinical diagnosis requires stable electrodes and high-efficient ECL signal amplification strategies. Herein, the authors discovered a new class of atomically dispersed peroxidase-like nanozymes with multiple active sites (CoNi-MOF@PCN-224/Fe), which significantly improved the catalytic performance and uncovered the underlying mechanism. Experimental studies and theoretical calculation results revealed that the nanozyme introduced a Fenton-like reaction into the catalytic system and the crucial synergistic effects of definite active moieties endow CoNi-MOF@PCN-224/Fe strong electron-withdrawing effect and low thermodynamic activation energy toward H₂O₂. Benefiting from the high peroxidase-like activity of the hybrid system, the resultant ECL electrode exhibited superior catalytic activity in the luminol-H₂O₂ system and resulted in an ≈17-fold increase in the ECL intensity. In addition, plasmonic Ag/Au core-satellite nanocubes (Ag/AuNCs) were designed as high-efficient co-reactant quenchers to improve the performance of the ECL immunoassay. On the basis of the differential signal amplification strategy (DSAS) proposed, the immunoassay displayed superior detection ability, with a low limit of detection (LOD) of 0.13 pg mL^-1 for prostate-specific antigen (PSA). The designed atomically anchored MOF-on-MOF nanozyme and DSAS strategy provides more possibilities for the ultrasensitive detection of disease markers in clinical diagnosis.