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Monoclonal antibody purification using cationic polyelectrolytes: an alternative to column chromatography
Thanmaya Peram1, Paul McDonald, Jayme Carter-Franklin
1Early Stage Purification, Process Research and Development, Genentech Inc., 1 DNA Way, South San Francisco, CA 94080, USA. peram.thanmaya@gene.com
Biotechnology Progress
|October 15, 2010
Summary
Cationic polyelectrolytes, like poly(arginine), can effectively precipitate impurities during monoclonal antibody purification, potentially replacing chromatography steps. This method offers consistent impurity clearance and antibody recovery across various conditions.
Area of Science:
- Biotechnology
- Chemical Engineering
- Biochemistry
Background:
- Monoclonal antibody (mAb) purification traditionally relies on chromatography.
- Developing alternative, cost-effective impurity removal methods is crucial.
- Cationic polyelectrolytes show promise for precipitating process-related impurities.
Purpose of the Study:
- Investigate cationic polyelectrolytes for impurity precipitation in mAb purification.
- Replace traditional chromatography steps with polyelectrolyte precipitation.
- Evaluate the impact of solution and polyelectrolyte properties on precipitation efficiency.
Main Methods:
- Studied precipitation using various cationic polyelectrolytes.
- Assessed effects of antibody isoelectric point, pH, ionic strength, and polyelectrolyte properties (molecular weight, pKa).
- Evaluated poly(arginine) precipitation as a flocculant and chromatography replacement.
Main Results:
- Increasing ionic strength reduced impurity precipitation at neutral pH.
- Higher polyelectrolyte molecular weight and pKa improved precipitation at higher ionic strength.
- Poly(arginine) demonstrated consistent host cell protein and DNA clearance, with high antibody recovery.
Conclusions:
- Poly(arginine) precipitation is a viable upstream alternative to anion exchange chromatography.
- This method achieves comparable impurity removal and antibody recovery.
- Cationic polyelectrolyte precipitation offers a scalable and efficient purification strategy.
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