Rapid purification of lysozyme by mixed-mode adsorption chromatography in stirred fluidized bed

The commercial STREAMLINE Direct HST adsorbent was employed to investigate the adsorption characteristics of lysozyme in complex chicken egg white (CEW). The effects of operating parameters, including adsorption pH, ionic strength, and hydrophobicity of liquids on binding capacity, were examined using a well-mixed contactor. To determine the elution strategy, the experiments were carried out in a small packed bed. The lysozyme was completely eluted by 0.5 M NaCl in 40 mM carbonate buffer (pH 12) at a high liquid velocity of 200 cm/h. The effect of rotating speed on fluidization characteristics was further investigated by using a stirred fluidized bed process. Smoother fluidization was observed when the rotating speed reached 200 rpm and lysozyme was directly recovered from highly viscous CEW in a single step with a high yield of 94.3% and a purification factor of 15.7.