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A facile and ingenious method to chemical etching-coordinating a metal-organic framework (MOF) followed by an annealing treatment was proposed to prepare Co₃O₄ nanoparticles uniformly dispersed in rational porous carbon nano-boxes (Co₃O₄@CNBs), which was further used to detect H₂O₂ released from living cells. The Co₃O₄@CNBs H₂O₂ sensor delivers much higher sensitivity than non-etching/coordinating Co₃O₄, offering a limit of detection of 2.32 nM. The wide working range covers 10 nM-359 <inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mi mathvariant="sans-serif">μ</mi></semantics></math></inline-formula>M H₂O₂, while possessing good selectivity and excellent reproducibility. Moreover, this biosensor was used to successfully real-time detect H₂O₂ released from living cells, including both healthy and tumor cells. The excellent performance holds great promise for Co₃O₄@CNBs’s applications in electrochemical biomimetic sensing, particularly real-time monitor H₂O₂ released from living cells.