Monolithic macroporous cryogel imprinted with protein G was prepared using a functional co-monomer of N-methacryloyl-L-phenylalanine and 2-hydroxyethyl methacrylate. The chemical structure of the cryogel prepared was studied by FTIR-spectroscopy and its porosity was analysed using scanning electron microscopy. The cryogel was used to purify protein G from recombinant Escherichia coli cell lysate and the effect of pH, temperature, ionic strength, flow rate, etc on the adsorption of protein G to the monolithic column have been investigated. The selectivity of the imprinted cryogel was studied using protein A and myoglobin. It was possible to capture about 9 mg of Protein G per g of the cryogel. (C) 2015 Elsevier B.V. All rights reserved.