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A High-throughput Automated Platform for the Development of Manufacturing Cell Lines for Protein Therapeutics
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High-throughput downstream process development for cell-based products using aqueous two-phase systems (ATPS) - A

Sarah Zimmermann1, Christian Scheeder1, Philipp K Zimmermann1

  • 1Karlsruhe Institute of Technology (KIT), Institute of Process Engineering in Life Science, Section IV: Biomolecular Separation Engineering (MAB), Karlsruhe, Germany.

Biotechnology Journal
|November 25, 2016
PubMed
Summary
This summary is machine-generated.

Aqueous two-phase systems (ATPS) offer a scalable method for cell purification. High-throughput screening and modeling effectively predict cell partitioning, aiding downstream process development for cell therapeutics.

Keywords:
Aqueous two-phase systemsCell cycle analysisCountercurrent distributionHigh-throughput screeningLabel-free cell separation

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Area of Science:

  • Biotechnology
  • Process Engineering
  • Cell Biology

Background:

  • Downstream processing of cell-based products is crucial for clinical development.
  • Aqueous two-phase systems (ATPS) provide a gentle, scalable, and cost-effective method for cell purification.
  • Robotic screening platforms enable high-throughput analysis of cell partitioning in ATPS.

Purpose of the Study:

  • To apply a robotic screening platform for high-throughput cell partitioning analysis in ATPS.
  • To design and validate a cell purification strategy using high-throughput screening (HTS) data and countercurrent distribution (CCD) modeling.
  • To investigate the influence of cell cycle phase and surface properties on cell separation.

Main Methods:

  • Utilized a robotic screening platform for high-throughput cell partitioning analysis in ATPS.
  • Developed a purification strategy based on HTS data and CCD modeling.
  • Experimentally validated the CCD model for cell purification.
  • Analyzed cell partitioning behavior of two model cell lines (HL-60 and L929).

Main Results:

  • Demonstrated excellent congruence between CCD model predictions and experimental data.
  • Confirmed that CCD models combined with HTS data are powerful tools for downstream process development.
  • Identified cell cycle phase as a significant factor influencing cell partitioning.
  • Determined that cell type-specific surface properties are the primary drivers of charge-dependent separation.

Conclusions:

  • The developed strategy using HTS and CCD modeling is effective for designing robust cell purification processes.
  • Cell surface properties play a key role in charge-dependent separation, while cell cycle phase also influences partitioning.
  • Maintaining constant growth conditions is advisable for robust purification process design.