Integrated continuous biomanufacturing on pilot scale for acid-sensitive monoclonal antibodies

Area of Science:

• Biotechnology
• Biopharmaceutical Manufacturing
• Process Engineering

Background:

• Acid-sensitive monoclonal antibodies (mAbs) are prone to aggregation at low pH, complicating their purification.
• Current bioprocessing methods often struggle to maintain the integrity of these sensitive molecules.

Purpose of the Study:

• To develop and demonstrate a pilot-scale integrated continuous bioprocess (ICB) for acid-sensitive mAbs.
• To ensure high recovery yields and purity while preventing antibody aggregation.

Main Methods:

• High cell density perfusion culture integrated with downstream processing.
• Downstream steps included Protein A chromatography (Zca ligand), virus inactivation, and ion-exchange chromatography.
• Mild pH conditions were maintained throughout the downstream process.
• Process integration and automation using Orbit software.

Main Results:

• Achieved high recovery yields (~90%) and productivity (~1 g/L/day).
• Maintained stable glycosylation patterns and efficiently removed impurities (host cell proteins, DNA).
• Negligible antibody aggregate levels were detected due to mild process conditions.

Conclusions:

• The developed ICB is suitable for purifying acid-sensitive mAbs at pilot scale.
• This work enables future industrial-scale continuous biomanufacturing of diverse antibody types.
• The mild conditions are key to preventing aggregation in sensitive biologics.