In this work, a chemiluminescent sensing paper has been developed using a peroxidase biomimetic metal−organic framework as a versatile host platform. For the first time, we have explored the use of in situ growth of Prussian Blue nanoparticles (PB-NPs) onto the MIL-101(Fe) structure for the assembly of a ready-to-use sensing paper. In situ growth of PB-NPs has been performed on the surface of the MIL-101(n) family. This novel composite, named PB-NPs@MIL-101(Fe), has been successfully used to develop a sensing paper for one-step detection of H 2 O 2 in real samples (commercial disinfectant solutions and tap water samples). The as-prepared material was fully characterized, including X-ray analysis, Fourier transform infrared, scanning and transmission electron microscopies, nitrogen isotherms, and elemental analysis. After the characterization, the analytical performance of the PB-NPs@MIL-101(Fe) sensing paper was evaluated. The low-cost sensor (0.15 euro per unit) was able to detect down to 8.2 μM (corresponding to 8.2 × 10 −11 mol) H 2 O 2 using only 10 μL of sample with satisfactory reproducibility (relative standard deviation 17%).