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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.

Sarah Zimmermann1, Sarah Gretzinger2, Marie-Luise Schwab3

  • 1Karlsruhe Institute of Technology (KIT), Institute of Process Engineering in Life Science, Section IV: Biomolecular Separation Engineering (MAB), Engler-Bunte-Ring 3, 76131 Karlsruhe, Germany.

Journal of Chromatography. A
|August 29, 2016
PubMed
Summary
This summary is machine-generated.

Automated screening of aqueous two-phase systems (ATPS) enables efficient cell separation for cell-based therapeutics. This high-throughput platform optimizes downstream processing, achieving high purity and recovery of specific cell populations.

Keywords:
Aqueous two-phase systems (ATPS)Downstream processing of cell-based productsHigh throughput screening (HTS)High-throughput flow cytometryLabel-free cell separation

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Area of Science:

  • Biotechnology
  • Cellular Biology
  • Process Engineering

Background:

  • Clinical development of cell-based therapeutics necessitates advanced downstream processing.
  • Aqueous two-phase systems (ATPS) offer label-free, scalable, and cost-effective cell separation.
  • High-throughput methods are crucial for optimizing ATPS for therapeutic applications.

Purpose of the Study:

  • To develop an automated robotic screening platform for high-throughput cell partitioning analysis in ATPS.
  • To investigate and optimize factors influencing cell separation in ATPS.
  • To assess the feasibility of separating specific cell populations for therapeutic applications.

Main Methods:

  • Development of an automated robotic screening system for ATPS.
  • High-throughput cell partitioning analysis using the HL-60 cell line.
  • Optimization of separation conditions and application of a counter-current distribution model.
  • Investigation of cell separation post-differentiation using DMSO.

Main Results:

  • The automated platform enables rapid and systematic investigation of ATPS parameters.
  • Optimized conditions were determined for HL-60 cell separation.
  • Successful separation of CD11b-positive and CD11b-negative HL-60 cells was demonstrated.
  • Modeling predicted >93% recovery of CD11b-positive cells with >99% purity using 30 transfers.

Conclusions:

  • The developed screening platform accelerates downstream process development for cell-based therapeutics.
  • ATPS, optimized via high-throughput screening, is a powerful tool for translational research.
  • This approach enables efficient purification of specific cell populations for therapeutic use.