A novel bifunctional MOF-encapsulated cobalt-doped carbon dots nanozyme (Co-CD/PMOF) with excellent peroxidasemimic catalytic activity and fluorescence property was synthesized and employed to fabricate a chemiluminescence/fluorescence (CL/FL) dualmode immunosensor for AFB1 detection. Co-CD/PMOF could catalyze the luminol/H 2 O 2 system to generate robust and long-lasting CL signals due to the slow diffusion effect and continuous generation of •OH, O 2•− , and 1 O 2 species. Differing from traditional flash-type CL emissions, this glow-type CL emission is helpful to fabricate a sensitive and accurate CL sensing platform. Then the CL/FL dual-mode detection of AFB1 was developed using antibody-functionalized Co-CD/PMOF as the signalamplifying nanoprobe. The CL mode assay based on indirect competitive immune principle was carried out on a chemiluminescence optical fiber platform, where the AFB1-OVA-functionalized optical fiber probe was employed for biorecognition, separation, and signal conducting. The AFB1 detection range and LOD were 0.63−69.36 ng/mL and 0.217 ng/mL, respectively. Using AFB1 antibodyfunctionalized immunomagnetic beads for capturing and separation, the FL mode detection of AFB1 was established based on the sandwich immune principle. A linear range of 0.54−51.91 ng/mL and a LOD of 0.027 ng/mL were obtained. This work designed a sensitive, rapid, and reliable nanozyme-powered dual-mode assay strategy and provided technical support in the field of environmental monitoring and food safety.