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Spin Filter Perfusion System for High Density Cell Culture: Production of Recombinant Urinary Type Plasminogen Activator in CHO Cells

Bio/Technology volume 8, pages 54–58 (1990)Cite this article

Abstract

We have used a 20 liter stirred tank fermentor, equipped with a 127 mesh ethylene-tetraflouroethylene rotating screen for cell recycle, for the continuous production of recombinant single chain urokinase-type plasminogen activator (rscu-PA) from Chinese hamster ovary (CHO) cells. Viable cell densities between 60 and 74 million per ml were maintained at medium perfusion rates of 3.0 to 4.0 fermentor volumes per day. Cells were retained by the 120 micron nominal opening filter through the formation of “clumped” cell aggregates of 200 to 600 μm in size, which did not foul the filter. In 31 days of culture, a total of 51 grams of rscu-PA were produced in 1,000 liters of medium. The rscu-PA produced over the course of this continuous culture was purified and characterized both in vitro and in vivo and shown to be comparable to natural scu-PA produced from the transformed human kidney cell line, TCL-598.

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Author information

Author notes

  1. George C. Avgerinos

    Present address: Tufts University, Talbot Ave., Medford, MA, 02155

  2. Denis Drapeau

    Present address: Genetics Institute, 1 Burtt Road, Andover, MA, 01810

  3. Jeff S. Socolow

    Present address: Applied Biotechnology Inc., 8 Rogers Street, Cambridge, MA, 02139

  4. Robert J. Broeze: Corresponding author

Authors and Affiliations

  1. Collaborative Research Inc., 2 Oak Park, Bedford, MA, 01730

    Jen-i Mao & Kathy Hsiao

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  1. George C. Avgerinos
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  2. Denis Drapeau
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  3. Jeff S. Socolow
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  4. Jen-i Mao
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  5. Kathy Hsiao
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  6. Robert J. Broeze
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Avgerinos, G., Drapeau, D., Socolow, J. et al. Spin Filter Perfusion System for High Density Cell Culture: Production of Recombinant Urinary Type Plasminogen Activator in CHO Cells. Nat Biotechnol 8, 54–58 (1990). https://doi.org/10.1038/nbt0190-54

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  • DOI: https://doi.org/10.1038/nbt0190-54

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