Rate parameters and gradient correlations of proteins are very important to engineering investigations of gradient-elution chromatography. Before carrying out computer simulations for gradient process, these rate parameters and gradient correlations must be determined from experimental data. This work presents a systematic method for estimating and determining these parameters and correlations using experimental data and computer-simulation results based on isocratic runs. The values determined were then employed in gradient-elution studies by putting them into the rate model to perform computer simulations. Experimental runs under gradient conditions using the parameters and correlations just referred to were carried out in a DEAE Sepharose CL-6B ion-exchange chromatography. Comparisons between the experimental data and the simulation results show good agreement.