Improved process design for monoclonal antibody charge variants separation with multicolumn counter-current solvent...

Area of Science:

• Biotechnology
• Chemical Engineering
• Chromatography

Background:

• Multicolumn counter-current solvent gradient purification (MCSGP) is effective for separating proteins with overlapping elution profiles.
• Discrepancies between batch process design and actual MCSGP conditions, due to changing flow rates and protein loads, reduce target protein purity and recovery.

Purpose of the Study:

• To develop an improved process design (reDesign) for MCSGP that generates more representative elution profiles.
• To enhance the separation performance of MCSGP by better mimicking continuous process conditions.

Main Methods:

• A novel reDesign strategy was implemented using the first-run MCSGP to capture actual continuous process elution profiles.
• The reDesign-based MCSGP (reMCSGP) was validated using a model protein mixture and monoclonal antibody (mAb) charge variants.

Main Results:

• For a model protein mixture, reMCSGP increased recovery from 83.6% to 97.8% while maintaining >95% purity.
• For mAb charge variants, reMCSGP achieved 93.9% recovery at >84% purity, outperforming standard MCSGP.

Conclusions:

• The reDesign strategy provides a more accurate elution profile that closely mirrors continuous process conditions.
• This enhanced process design significantly improves the separation performance, purity, and recovery in MCSGP.