Development Of A Two-Column Antibody Downstream Process

The current standard monoclonal antibody purification process is typically composed of three column steps – protein A chromatography, then cation exchange chromatography, followed by anion exchange chromatography or multimodal chromatography. Protein A serves as a robust capture step, but it is also a key purification step. various washing conditions were investigated with a DOE based approach to optimize protein A chromatography. In this study, addition of Triton X-100 to the wash buffers was explored to reduce impurities. In addition, the optimized protein A column method was also shown to be a robust viral clearance step. A second CEX chromatography step such as Poros XS® was also optimized with a DOE based approach. In summary, a two-column downstream process for some antibodies was developed instead of a three-column process, offering some advantages including reduced manufacturing time, reduced cost of goods, and manufacturing simplicity.