The separation of two similarly sized proteins, bovine serum albumin (BSA) and bovine hemoglobin (Hb) was carried out using a new type of ion-exchange mixed-matrix adsorber membranes. The adsorber membranes were prepared by incorporation of various types of Lewatit ion-exchange resins into an ethylene-vinyl alcohol copolymer porous structure. The obtained heterogeneous matrices, composed of solid particles surrounded by the polymeric film, display high static and dynamic protein adsorption capacities. The effect of operational parameters such as filtration flow-rate, pH, and ionic strength on the protein separation performances was investigated for cation- as well as anion-exchange adsorber membranes. An average separation factor was calculated by numerical integration of the protein concentration in the permeate curve during the filtration run. High average separation factor values were obtained for BSA-Hb separation at physiological ionic strength with a filtration flow-rate up to 20 l/h per m2, until the protein breakthrough point at 10% of the feed concentration.