Alternative To A Three-Step Monoclonal Antibody Purification Process: A Mixed-Mode Approach

Monoclonal antibodies (mAbs) continue to increase in prevalence, resulting in manufacturers striving to improve their purification process speed and efficiency. Purification of mAbs typically consists of three steps, involving a Protein A-based capture followed by intermediate and final polishing steps. Mixed Mode chromatography has become a topic of discussion for manufacturer looking to reduce the number of steps in their mAb purification processes.

Three purification processes were recently compared to determine their strengths and shortcomings.

• A three-step mAb purification process using IEX resins following Protein A capture
• A two-step mAb process using a hydrophobic anion exchange (AEX) mixedmode resin
• And a two-step approach using a calcium affinity cation exchange (CEX) mixed-media

These processes were evaluated for yield and purity of mAb purification and demonstrate that a mAb can be effectively purified with high yield and purity via two-step workflows with different types of mixed-mode resins.