Developing　nanozyme-based　free　radical　scavenging　is　a　promising　signal　modulation　approach　for　ECL　sensing.　Nevertheless,　the　relatively　low　antioxidant　activity　and　inherent　pro-oxidant　activity　of　numerous　nanozymes　have　significantly　hindered　the　development　of　this　strategy.　Here　a　biofunctional　copper-based　metal-organic　framework　(CuMOF)　with　multiple　enzyme-mimicking　activities　was　employed　for　the　modulation　of　the　ECL　immunosensor,　guided　by　the　self-cascade　antioxidant　reaction.　The　inherent　SOD,　CAT,　and　the　capacity　to　eliminate　·OH　endow　CuMOF　with　powerful　synergistic　antioxidant　effects　while　little　pro-oxidant　activities　were　displayed,　enabling　efficient　scavenging　of　the　O2·-　produced　during　the　electrochemical　oxidation　of　H2O2.　Subsequently,　the　nanoconfinement　effect　of　the　layered　double　hydroxide　was　introduced　to　ensure　a　steady　ECL　signal.　The　suggested　ECL　immunosensor,　using　aflatoxin　B1　as　a　proof-of-concept　target,　demonstrated　a　detection　range　spanning　from　0.001　pg/mL　to　10　ng/mL,　with　the　detection　limit　calculated　to　be　0.18　fg/mL.　This　exceptional　achievement　greatly　broadens　the　range　of　possible　uses　for　nanozyme-based　radical　scavenging　modulated　ECL　analysis.　©　2024　American　Chemical　Society.