Lysozyme adsorption and purification by expanded bed chromatography with a small-sized dense adsorbent

Lysozyme adsorption and purification by expanded bed chromatography of a customized Nd-Fe-B alloy-densified agarose (NFBA) gel modified with Cibacron Blue 3GA (CB) were investigated, and the results were compared with those obtained with CB-modified Streamline gel (CB-Streamline). The NFBA gel had a mean size of 102 mum and a mean density of 1.88 g/ml. The breakthrough behavior of lysozyme was modeled considering the dispersion of the liquid and solid phases and the diffusive mass transport of protein to the solid phase. Using independently determined parameters, the model prediction agreed reasonably well to the experimental data. Although the two dye-ligand adsorbents showed nearly the same static capacity, the dynamic binding capacity of the CB-NFBA gel was nearly twice that of the CB-Streamline gel. Moreover, lysozyme was purified from chicken egg white solution by the expanded beds with the two adsorbents, and the expanded bed with the CB-NFBA gel produced much larger purification factor than that with the CB-Streamline gel. All the results indicated that the small-sized dense medium CB-NFBA gel was more efficient as an expanded bed adsorbent. (C) 2002 Elsevier Science B.V. All rights reserved.