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Related Concept Videos

Downstream Processing01:29

Downstream Processing

Downstream processing begins once fermentation is complete and involves a series of steps to recover and purify products such as acids, vitamins, antibiotics, or proteins.Cell HarvestingFor example, for intracellular protein-based products, the first step is harvesting the cells. This is typically achieved using centrifugation or filtration to separate the cells from the liquid phase.Cell Disruption for Intracellular ProductsIf the target product is intracellular, the harvested cells must be...
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Downstream antibody purification using aqueous two-phase extraction.

Lisong Nathan Mao1, Jameson K Rogers, Matthew Westoby

  • 1Purification Process Development, Biogen IDEC Corporation, 5200 Research Place, San Diego, CA 92122, USA. nathan.mao@biogenidec.com

Biotechnology Progress
|September 21, 2010
PubMed
Summary
This summary is machine-generated.

Polyethylene glycol (PEG)-citrate aqueous two-phase systems (ATPS) effectively extract monoclonal antibodies (mAbs). Optimization using design of experiments (DOE) significantly reduced host cell proteins (HCPs) and impurities, enhancing antibody purity and recovery.

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Area of Science:

  • Bioseparations Engineering
  • Protein Purification
  • Biotechnology

Background:

  • Antibody production relies on efficient purification methods.
  • Aqueous two-phase systems (ATPS) offer a promising approach for bioseparations.
  • Polyethylene glycol (PEG)-citrate ATPS are explored for antibody extraction.

Purpose of the Study:

  • To investigate the extraction of monoclonal antibodies (mAbs) using PEG-citrate ATPS.
  • To identify optimal conditions for antibody partitioning and impurity removal.
  • To develop a scale-up model for continuous extraction processes.

Main Methods:

  • Utilized purified monoclonal antibody (mAb) studies to define operating ranges.
  • Employed statistical design of experiments (DOE) to analyze antibody and host cell protein (HCP) separation.
  • Applied a Steepest Descent algorithm for optimizing HCP partitioning and purity.

Main Results:

  • Antibody partitioning was nearly complete across examined operating conditions.
  • Optimal ATPS (14.0% PEG, 8.4% citrate, 7.2% NaCl, pH 7.2) yielded 89% recovery and 7.6-fold HCP reduction.
  • A separate system (15% PEG, 8% citrate, 15% NaCl, pH 5.5) reduced impurities to <0.5% with 95% recovery.

Conclusions:

  • PEG-citrate ATPS are effective for high-purity antibody extraction from cell culture media.
  • Optimized conditions significantly improve antibody purity and reduce host cell proteins.
  • A scale-up model for counter-current extraction suggests potential for continuous processing improvements.