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Upstream processing represents a critical phase in biomanufacturing, wherein biological systems such as microorganisms, mammalian cells, or insect cells are cultivated to produce therapeutic proteins, vaccines, enzymes, or other biologically derived products. This phase encompasses all steps from the selection and genetic manipulation of the production organism to the cultivation of cells in bioreactors under tightly controlled environmental conditions.Host Selection and Genetic OptimizationThe...

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A Microfluidic Platform for High-throughput Single-cell Isolation and Culture
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Microwell engineering characterization for mammalian cell culture process development.

Timothy A Barrett1, Andrew Wu, Hu Zhang

  • 1Department of Biochemical Engineering, University College London, UK.

Biotechnology and Bioengineering
|September 10, 2009
PubMed
Summary
This summary is machine-generated.

Shaken microwell plates offer a cost-effective platform for cell culture optimization, yielding results comparable to shake flasks. This technology reduces scale and material needs for high-throughput cell line evaluation.

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Area of Science:

  • Biotechnology
  • Bioprocess Engineering
  • Cell Culture Technology

Background:

  • Shaken microplate formats present a potential platform for rapid cell culture optimization.
  • Microwell systems could provide early process design data cost-effectively with reduced material requirements if kinetics match shake flasks.

Purpose of the Study:

  • To engineer and characterize liquid mixing and gas-liquid mass transfer in microwell systems.
  • To assess the impact of these parameters on suspension cell culture (murine hybridoma producing IgG1).

Main Methods:

  • Engineering characterization of 24-well plates using Computational Fluid Dynamics (CFD) for energy dissipation (P/V) and shear rate.
  • Quantification of mixing time via iodine decolorization experiments.
  • Measurement of oxygen transfer rate (k(L)a) as a function of shaking frequency and liquid fill volume.

Main Results:

  • Predicted P/V ranged from 5 to 35 W m(-3); k(L)a values varied between 1.3 and 29 h(-1); mixing times ranged from 1.7 s to 3.5 h.
  • High shaking speeds (>250 rpm) negatively impacted hybridoma cell growth.
  • Oxygen-limited conditions occurred at low shaking speeds with high fill volumes (120 rpm, 2,000 microL).
  • Cell growth kinetics and antibody titers were comparable between microwell plates and shake flasks at matched energy dissipation rates.

Conclusions:

  • Shaken microwell plates provide reproducible and comparable data to shake flask systems.
  • This approach offers a significant decrease (≥30-fold) in scale and material requirements.
  • The technology, coupled with automation, enables high-throughput evaluation of cell lines under realistic suspension culture conditions.