Molecularly imprinted thin membranes for selective removal of tetracycline from real water samples were prepared using the radiation-induced polymerization method. The chemical and physical characterization of the membranes was conducted by FTIR, XPS, AFM and SEM analyses. The effect of the template on the size and the size distribution of the pores in the membrane structure was investigated by positron annihilation lifetime spectroscopy (PALS) experiments. The presence of the template molecule causes the formation of larger cavities, which have a strong correlation to the molecular size of the template molecule. The density of the free volume holes in the imprinted network shows a significant increase due to the presence of the template molecule. The binding performances of the membranes were tested against various factors such as pH, time and initial concentration of template molecules. The specific selectivity of the membranes was investigated by cross-reactivity experiments. The binding capacity of the imprinted membranes was obtained at 14.5 mu mol g(-1), with the highest imprinting factor (2.84) for tetracycline. The imprinting factor was obtained at 1.44, 1.25 and 1.57 for oxytetracycline hydrochloride, doxycycline hyclate and chlortetracycline, respectively. The binding capability of the imprinted membranes in real water samples was promising for the application of the membranes as a filter material for removal of tetracycline with high binding capacity. Tetracycline binding percentage of the membranes was determined at 92.4, 81.1., 75.0 and 68.7 for ultra-pure water, tap water and natural water samples collected from different sources, respectively.