Fluorinert, an oxygen carrier, improves cell culture performance in deep square 96-well plates by facilitating oxygen transfer

Authors

  • Aaron Meyer,

    1. Protein Expression Technologies, Biotechnology Process Development, Merck Research Laboratories, 1011 Morris Avenue, Union, NJ 07083
    Current affiliation:
    1. Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 01239.
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    • Aaron Meyer and Russell G. G. Condon contributed equally to the project

  • Russell G. G. Condon,

    1. Protein Expression Technologies, Biotechnology Process Development, Merck Research Laboratories, 1011 Morris Avenue, Union, NJ 07083
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    • Gregory Keil,

      1. Protein Expression Technologies, Biotechnology Process Development, Merck Research Laboratories, 1011 Morris Avenue, Union, NJ 07083
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    • Nikita Jhaveri,

      1. Protein Expression Technologies, Biotechnology Process Development, Merck Research Laboratories, 1011 Morris Avenue, Union, NJ 07083
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    • Zhong Liu,

      1. Protein Expression Technologies, Biotechnology Process Development, Merck Research Laboratories, 1011 Morris Avenue, Union, NJ 07083
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    • Yung-Shyeng Tsao

      Corresponding author
      1. Protein Expression Technologies, Biotechnology Process Development, Merck Research Laboratories, 1011 Morris Avenue, Union, NJ 07083
      • Protein Expression Technologies, Biotechnology Process Development, Merck Research Laboratories, 1011 Morris Avenue, Union, NJ 07083
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    Abstract

    In bioprocess development, the 96-well plate format has been widely used for high-throughput screening of production cell line or culture conditions. However, suspension cell cultures in conventional 96-well plates often fail to reach high cell density under normal agitation presumably due to constraints in oxygen transfer. Although more vigorous agitation can improve gas transfer in 96-well plate format, it often requires specialized instruments. In this report, we employed Fluorinert, a biologically inert perfluorocarbon, to improve oxygen transfer in 96-well plate and to enable the growth of a Chinese Hamster Ovary cell line expressing a recombinant monoclonal antibody. When different amounts of Fluorinert were added to the cell culture medium, a dose-dependent improvement in cell growth was observed in both conventional and deep square 96-well plates. When sufficient Fluorinert was present in the culture, the cell growth rate, the peak cell density, and recombinant protein production levels achieved in deep square 96-wells were comparable to cultures in ventilated shake flasks. Although Fluorinert is known to dissolve gases such as oxygen and CO2, it does not dissolve nor extract medium components, such as glucose, lactate, or amino acids. We conclude that mixing Fluorinert with culture media is a suitable model for miniaturization of cell line development and process optimization. Proper cell growth and cellular productivity can be obtained with a standard shaker without the need for any additional aeration or vigorous agitation. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2012

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