Novel micro-bioreactor high throughput technology for cell culture process development: Reproducibility and scalability assessment of fed-batch CHO cultures

Authors

  • Ashraf Amanullah,

    Corresponding author
    1. Vaccine & Biologics Process Development, Merck Research Laboratories, Merck & Co. Inc. West Point, Pennsylvania, 19486
    Current affiliation:
    1. Genentech Inc., I Antibody Way, Oceanside, CA 92056. telephone: 760-231-3050; fax: 760-231-2465.
    • Vaccine & Biologics Process Development, Merck Research Laboratories, Merck & Co. Inc. West Point, Pennsylvania, 19486.
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  • Jose Manuel Otero,

    1. Vaccine & Biologics Process Development, Merck Research Laboratories, Merck & Co. Inc. West Point, Pennsylvania, 19486
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  • Mark Mikola,

    1. Vaccine & Biologics Process Development, Merck Research Laboratories, Merck & Co. Inc. West Point, Pennsylvania, 19486
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  • Amy Hsu,

    1. Bioprocess R&D, Merck Research Laboratories, Merck & Co. Inc., Rahway, New Jersey 07065
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  • Jinyou Zhang,

    1. Vaccine & Biologics Process Development, Merck Research Laboratories, Merck & Co. Inc. West Point, Pennsylvania, 19486
    Current affiliation:
    1. Immunomedics, Inc., 300 American Road, Morris Plains, NJ 07950.
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  • John Aunins,

    1. Vaccine & Biologics Process Development, Merck Research Laboratories, Merck & Co. Inc. West Point, Pennsylvania, 19486
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  • H. Brett Schreyer,

    1. Seahorse Bioscience Inc., 16 Esquire Road, Billerica, Massachusetts 01862
    Current affiliation:
    1. Genomatica, Inc., 10520 Wateridge Circle, San Diego, CA 92121.
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  • James A. Hope,

    1. Seahorse Bioscience Inc., 16 Esquire Road, Billerica, Massachusetts 01862
    Current affiliation:
    1. JHope Bioprocess Consultants, PO Box 81, Accord, MA 02018.
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  • A. Peter Russo

    1. Seahorse Bioscience Inc., 16 Esquire Road, Billerica, Massachusetts 01862
    Current affiliation:
    1. Merck & Co. Inc., Rahway, NJ 07065.
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Abstract

With increasing timeline pressures to get therapeutic and vaccine candidates into the clinic, resource intensive approaches such as the use of shake flasks and bench-top bioreactors may limit the design space for experimentation to yield highly productive processes. The need to conduct large numbers of experiments has resulted in the use of miniaturized high-throughput (HT) technology for process development. One such high-throughput system is the SimCell™ platform, a robotically driven, cell culture bioreactor system developed by BioProcessors Corp. This study describes the use of the SimCell™ micro-bioreactor technology for fed-batch cultivation of a GS-CHO transfectant expressing a model IgG4 monoclonal antibody. Cultivations were conducted in gas-permeable chambers based on a micro-fluidic design, with six micro-bioreactors (MBs) per micro-bioreactor array (MBA). Online, non-invasive measurement of total cell density, pH and dissolved oxygen (DO) was performed. One hundred fourteen parallel MBs (19 MBAs) were employed to examine process reproducibility and scalability at shake flask, 3- and 100-L bioreactor scales. The results of the study demonstrate that the SimCell™ platform operated under fed-batch conditions could support viable cell concentrations up to least 12 × 106 cells/mL. In addition, both intra-MB (MB to MB) as well as intra-MBA (MBA to MBA) culture performance was found to be highly reproducible. The intra-MB and -MBA variability was calculated for each measurement as the coefficient of variation defined as CV (%) = (standard deviation/mean) × 100. The % CV values for most intra-MB and intra-MBA measurements were generally under 10% and the intra-MBA values were slightly lower than those for intra-MB. Cell growth, process parameters, metabolic and protein titer profiles were also compared to those from shake flask, bench-top, and pilot scale bioreactor cultivations and found to be within ±20% of the historical averages. Biotechnol. Bioeng. 2010; 106: 57–67. © 2010 Wiley Periodicals, Inc.

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