An electrochemical impedimetric immunosensor was constructed based on a hierarchical nanostructured CoCo Prussian blue analogue entrapped by a Zr-based porphyrin MOF (denoted by [email protected]) for the sensitive detection of deoxynivalenol (DON). Given that [email protected] demonstrated a hierarchical nanostructure, large specific surface area, and the synergistic effect between mixed metal valence states (Co2+/Co3+) and Zr clusters, it thus displayed a high binding interaction with the DON-targeted antibody. Compared with CoCoPBA and PCN-221, [email protected] showed a superior stabilization ability toward the antibody-antigen complex in aqueous solution. Under optimum conditions, the [email protected] impedimetric immunosensor exhibited a good linear range from 1 fg mL−1 to 1 ng mL−1, and an ultralow detection limit of 0.14 fg mL−1, accompanied by high selectivity, reproducibility, stability, and applicability in foodstuffs. This method allows the integration of MOFs with cascading properties for applications in the biosensing and food safety fields.