Application Note

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

Antibodies And Proteins GettyImages-1364886992

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.

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