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Biosensor stability and performance are known to benefit from the nanoscale confinement of its immobilized protein or enzyme components due to its ability to mimic the natural environment of these biomolecules. In this study, the enzyme cholesterol oxidase (ChOx) was adsorbed on pores created on polystyrene-block-polymethyl methacrylate films. A combination of fluorescence microscopy and atomic force microscopy analysis reveals the presence of ChOx on the polystyrene domain pores. Absorbance measurements on a cholesterol solution exposed to the enzyme deposited on the block copolymer reveal the occurrence of catalysis. The results of this study demonstrate the feasibility of immobilizing enzymes in a polymer-derived bioinspired platform that can be utilized for biosensing applications.