Paper-based sensors open a new avenue to produce easy-to-handle, inexpensive and simple-use analytical devices for widespread application fields in biological systems involving diagnosis, drug discovery and biosensing. In this manuscript, we constructed a fluorescence sensor based on functionalized paper coupled with silver nanoparticles (AgNPs) etching for hydrogen peroxide (H₂O₂) determination in biological systems. A significant fluorescence signal was achieved by immobilizing CDs onto the surface of cellulose filter paper by covalent bonds. The CDs functionalized paper coupled with AgNPs composites platform (PCD/AgNPs) consisted of CDs functionalized paper (PCD) and a AgNPs layer which acted as a fluorescence quencher of CDs via a physical adsorption mechanism. With the introduction of H₂O₂, the AgNPs were etched accompanied by the fluorescence recovery. As the target, a good linear relationship of a fluorescence turn-on response toward H₂O₂ between 10 and 120 μM was obtained with a limit of detection of 1.38 μM in our sensor. The designed sensor demonstrated good specificity and would have broad application prospects in the detection of milk samples and determination of H₂O₂ released from cancer cells.