Expanded bed adsorption (EBA) is a special chromatography technique with perfect classification of adsorbent particles in the column, thus the performance of protein adsorption in expanded beds is particular, obviously nonuniform and complex along the column. Detailed description of the complex adsorption kinetics of proteins in expanded bed is essential for better analyzing of adsorptive mechanisms, the design of chromatographic processes and the optimization of operation parameters of EBA processes. In this work, a theoretical model for the prediction of protein adsorption kinetics in expanded beds was developed by taking into account the classified distribution of adsorbent particles along the bed height, the nonuniform behaviors of axial liquid dispersion, the axial variation of local bed voidage as well as the axial changes of target component mass transfer. The model was solved using the implicit finite difference scheme combining with the orthogonal collocation method, and then applied to predict the breakthrough behaviors of bovine serum albumin (BSA) on StreamlineDEAEand lysozyme on Streamline SP along the bed height in expanded beds under various conditions. In addition, the experiments of front adsorption of BSA on Streamline DEAE at different axial column positions were carried out to reveal the adsorption kinetics of BSA along the bed height in a 20mm I.D. expanded bed, and the influences of liquid velocity and feed concentration on the breakthrough behaviors were also analyzed. The breakthrough behaviors predicted by the present model were compared with the experimental data obtained in this work and in the literature published. The agreement between the prediction and the experimental breakthrough curves is satisfied.