Using polyurethane (PU) as the carrier, membrane protein as the template, NVinylformamide (NVF) as the monomer, and 1,4-Butanediol diglycidyl ether (BDDE) as the crosslinker, in the presence of gelatin, and initiated by ultraviolet radiation, the hydrogel PUNVF-Gelatin was prepared. The effects of monomer concentration, crosslinker concentration, and gelatin concentration on the adsorption capacity for the membrane protein and the imprinting efficiency were studied. The results showed that the maximum adsorption capacity for the membrane protein on the PU-grafted imprinted hydrogel was achieved when the monomer mass fraction was 5%, the crosslinker mass fraction was 3%, and the gelatin mass fraction was 0.6%.