The breakthrough curve performance of lysozyme and bovine serum albumin to immobilized Cibacron 3GA using different solid supports such as gel beads and membrane disks was investigated in this work. The effects of flow rate and different module designs were also studied. Variation in the flow rate was found to be insignificant for the column process in both nonadsorption and single-protein experiments, but it affected the elution peak height for membrane disks. The peak height decreased with increasing flow rate. As for the effect of different designs, a long column and a wide membrane stack induced a broader breakthrough performance. The performance varied with different solid supports in two-protein experiments. Competitive adsorption occurred for a gel bead column, and the breakthrough curve performance resembled the prediction of combining local equilibrium theory and the extra module effect. The affinity strenth to the gel bead support is in the order lysozyme > BSA dimer > BSA monomer. As for membrane performance, two BSA solutes did not adsorb onto membranes, so as simple separation of lysozyme from BSA, instead of displacement phenomena, was observed.