Displacement Chromatography of Proteins Under Elevated Flow Rate and Crossing Isotherm Conditions

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Abstract
Displacement chromatography was employed for the preparative‐scale separation of four‐component protein mixtures. The feed components were both purified and concentrated during the separation process. Protein displacement was successfully carried out at elevated flow rates with minimal effect on bioproduct purity resulting in high bioproduct throughput. The effect of “crossing” adsorption isotherms on the displacement chromatographic process was investigated. Under adsorptive conditions where the protein isotherms cross, the displacement system was unable to separate the feed components. On the other hand, subtle changes in the adsorption conditions readily “separated” the protein isotherms, enabling the purification of the proteins of interest by displacement chromatography. This paper extends the scope of biopolymer displacement chromatography to more complex feed mixtures and elevated flow rates, and demonstrates how protein isotherms can be manipulated to effect the efficient purification of biopolymers by displacement chromatography.