In this study, we introduced a new strategy to design interface imprinted polymers for a novel aspect of molecular imprinting technique, utilization of sacrificial metal oxide particles. In the first step, bovine hemoglobin (BHb) was adsorbed on zinc oxide (ZnO) particles, which were then used to synthesize polyacrylic acid-based molecular imprinting membrane by bulk polymerization in the presence of ethylene glycol dimethacrylate as a cross-linking agent. After polymerization terminated, BHb-ZnO particles were removed to leave effective imprint sites onto the bulk polymeric network which is responsible for the formation of template orientation. The characterization of membranes was investigated by using Fourier transform infrared (FTIR), Raman spectroscopy (RS), scanning electron microscopy (SEM), surface area measurements (BET analyses) and thermogravimetric analysis (TGA). The interface molecularly imprinted membranes (iMIMs) have a relatively high specific rebinding capacity of 65.98 mg/g and excellent selectivity towards BHb with a separation factor of 6.78. The equilibrium adsorption isotherms fitted well to Langmuir isotherms (R-2 = 0.9944) and the value of adsorption capability (Q(max)) and equilibrium constant (b) were estimated to be 73.53 mg/g and 1.36 mg/mL for the iMIM, respectively. The kinetics of adsorption fitted best to pseudo-second order (R-2 = 0.9912). The ZnO particles were used not only to ensure the preservation of the imprint cavities in the polymer network but also to lead to high template removal and better rebinding kinetics. This novel design with multiple recognition sites is quite simple and suitable for the separation of biomacromolecules.