Performance optimization of continuous countercurrent tangential chromatography for antibody capture.

Area of Science:

• Biotechnology
• Chemical Engineering
• Bioprocess Engineering

Background:

• Conventional packed bed protein A chromatography has limitations for monoclonal antibody purification.
• Continuous countercurrent tangential chromatography (CCTC) presents a promising alternative for antibody purification.
• Optimization of CCTC is crucial for industrial applications.

Purpose of the Study:

• To optimize continuous countercurrent tangential chromatography (CCTC) for monoclonal antibody purification.
• To evaluate the impact of various parameters on CCTC performance, including yield, impurity removal, productivity, and buffer usage.
• To provide a framework for designing and optimizing CCTC systems.

Main Methods:

• Process development and CCTC experiments using bench-scale data.
• Modeling CCTC for protein A capture of monoclonal antibodies from Chinese Hamster Ovary cell culture feedstocks.
• Assessment of resin binding capacity, kinetics, staging strategy, and buffer recycling.

Main Results:

• Optimal staging in the binding step improved yield and increased system productivity by 8-16%.
• Increased stages in wash/elution steps reduced buffer usage by ~40% and improved impurity removal by ~2 logs.
• Buffer recycling in wash/regeneration steps further reduced buffer usage by ~35%.
• Smaller particle size resins increased CCTC productivity by 2.5-fold (up to 190 g mAb/L resin/hr).

Conclusions:

• CCTC optimization significantly enhances monoclonal antibody purification efficiency.
• Strategic staging, buffer recycling, and resin selection are key to maximizing CCTC performance.
• Optimized CCTC provides a robust framework for industrial-scale antibody capture applications.