(312e) Enhancing Affinity Chromatography Resin Efficiency Using 3 Column Periodic Continuous Chromatography

Protein A Affinity Chromatography is widely used for purification of monoclonal antibodies from harvested cell culture fluid (HCCF). The HCCF is typically loaded on a single large Protein A affinity chromatography column in cycles until all of the HCCF is processed. Protein A resin costs are significant and can be reduced by operating the process continuously using multiple columns in lieu of one large column. In this study, a series of experiments were performed using three 1ml Mab Select SuRe columns on a modified AKTA system (3C PCC) from GE Healthcare. The columns were loaded individually until the 70% breakthrough point was achieved. The non-bound fluid from the column was then loaded onto the next column to capture the Mab, preventing any protein loss. At any given point, all three columns were in operation; either loading or washing, enabling a reduction in processing time. The product yield and quality were evaluated and compared to current process to determine the affect of using the continuous process. The continuous operation shows the potential to reduce both resin volume and buffer consumption, but the system hardware and the process are more complex than the current process. However, cost savings may make it an attractive option for both new and existing processes.