Molecularly imprinted polymers possessing synthetic recognition sites present great potential for various application areas, which require specific recognition of molecules. However various molecularly imprinted polymers (MIPs) reported previously have low affinity capacity and low binding kinetics due to weak interactions between template molecule and functional monomer in aqueous media and also high mass transfer resistance. The synthesis of MIPs onto nanomaterial surface via metal ion coordination interactions serves as an effective solution. In this work, we prepared thin molecularly imprinted polymer layer on bacterial cellulose nanofiber surface using Cu2+ chelating monomer, N-Methacryloyl-(L)-histidinemethylester (MAH) as a functional monomer orienting cytochrome c molecules. The conditions of polymerization and adsorption were optimized to obtain best affinity performance. Due to unique properties of bacterial cellulose nanofibers such as high surface-to-volume ratio and macro-porosity, the resultant cytochrome c imprinted nanofibers (Cyt c-MIP NFs) reducing diffusional resistances could achieve high recognition capacity and excellent binding kinetics. Cyt c-MIP NFs were also successfully applied to capture template proteins from real sample. This novel procedure presents simple, cheap and stable fabrication strategy for efficient separation of proteins. (C) 2015 Published by Elsevier Ltd.