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Accelerating perfusion process optimization by scanning non-steady-state responses

Authors

  • Sumitra Angepat,

    1. Michael Smith Laboratories, University of British Columbia, 2185 East Mall, Vancouver, British Columbia V6T 1Z4, Canada
    2. Department of Chemical and Biological Engineering, University of British Columbia, 2216 Main Mall, Vancouver, British Columbia V6T 1Z4, Canada
    Current affiliation:
    1. Abbott Bioresearch Center, 100 Research Drive, Worcester, MA 01605.
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  • Volker M. Gorenflo,

    1. Michael Smith Laboratories, University of British Columbia, 2185 East Mall, Vancouver, British Columbia V6T 1Z4, Canada
    2. Department of Chemical and Biological Engineering, University of British Columbia, 2216 Main Mall, Vancouver, British Columbia V6T 1Z4, Canada
    Current affiliation:
    1. Sanofi Pasteur, 1755 Steeles Avenue West, Toronto, ON M2R 3T4, Canada.
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  • James M. Piret

    Corresponding author
    1. Michael Smith Laboratories, University of British Columbia, 2185 East Mall, Vancouver, British Columbia V6T 1Z4, Canada
    2. Department of Chemical and Biological Engineering, University of British Columbia, 2216 Main Mall, Vancouver, British Columbia V6T 1Z4, Canada
    • Michael Smith Laboratories, University of British Columbia, 2185 East Mall, Vancouver, British Columbia V6T 1Z4, Canada; telephone: 604-822-5835; fax: 604-822-2114
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Abstract

Perfusion processes provide consistent culture conditions, high productivity and low product residence times. However, process development can be slow due to the 1 week or more required to reach each steady state. The objective of this work was to accelerate process development in perfusion cultures by scanning non-steady-state transient responses to qualitatively predict steady-state performance. The method was tested using a shift in temperature every 3 days, scanned down by steps of 2°C from 37°C to 31°C, then scanned up to 37°C. Higher t-PA concentrations were predicted at lower temperatures, confirmed by subsequent pseudo-steady-state results. In most cases, transient values on the 3rd day were in close concordance with pseudo-steady-state values. To further accelerate process development, transient scanning was applied to small-scale, non-instrumented cultures. Similar results were obtained, although quantitative t-PA values were 15–30 times lower than in high cell density perfusion cultures. The method was further explored by investigating 1 day transient shifts in temperature where more variability was observed, suggesting that the cells were still adapting to the new environment. Nonetheless, the overall response again qualitatively predicted the pseudo-steady-state temperature response. Use of transient scanning in conjunction with pseudo-steady-state verification and refinement of optimal results could reduce process development time to a third or less of comparable steady-state-based optimization.

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