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.