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Automation of high CHO cell density seed intensification via online control of the cell specific perfusion rate and its impact on the N-stage inoculum quality.

Authors
  • Schulze, Markus1
  • Lemke, Johannes2
  • Pollard, David3
  • Wijffels, Rene H4
  • Matuszczyk, Jens2
  • Martens, Dirk E5
  • 1 Corporate Research, Sartorius Stedim Biotech GmbH, August-Spindler-Str. 11, 37079, Göttingen, Germany; Bioprocess Engineering, Wageningen University, PO Box 16, 6700 AA, Wageningen, The Netherlands. Electronic address: [email protected] , (Germany)
  • 2 Corporate Research, Sartorius Stedim Biotech GmbH, August-Spindler-Str. 11, 37079, Göttingen, Germany. , (Germany)
  • 3 Corporate Research, Sartorius Stedim North America, 6 Tide Street, Boston MA, 02210, United States. , (United States)
  • 4 Bioprocess Engineering, Wageningen University, PO Box 16, 6700 AA, Wageningen, The Netherlands; Biosciences and Aquaculture, Nord University, N-8049 Bodø, Norway. , (Netherlands)
  • 5 Bioprocess Engineering, Wageningen University, PO Box 16, 6700 AA, Wageningen, The Netherlands. , (Netherlands)
Type
Published Article
Journal
Journal of biotechnology
Publication Date
Jul 20, 2021
Volume
335
Pages
65–75
Identifiers
DOI: 10.1016/j.jbiotec.2021.06.011
PMID: 34090946
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Current CHO cell production processes require an optimized space-time-yield. Process intensification can support achieving this by enhancing the productivity and improving facility utilization. The use of perfusion at the last stage of the seed train (N-1) for high cell density inoculation of the fed-batch N-stage production culture is a relatively new approach with few industry applicable examples. Within this work, the impact of the cell-specific perfusion rate (CSPR) of the N-1 perfusion and the relevance of its control for the quality of generated inoculation cells was evaluated using an automated perfusion rate (PR) control based on online biomass measurements. Precise correlations (R² = 0.99) between permittivity and viable cell counts were found up to the high densities of 100⋅106 c·mL-1. Cells from N-1 perfusion were cultivated at a high and low CSPR with 50 and 20 pL·(c·d)-1, respectively. Lowered cell growth and an increased apoptotic reaction was found as a consequence of the latter due to nutrient limitations and reduced uptake rates. Subsequently, batch cultivations (N-stage) from the different N-1 sources were inoculated to evaluate the physiological state of the inoculum. Successive responses resulting from the respective N-1 condition were uncovered. While cell growth and productivity of approaches inoculated from high CSPR and a conventional seed were comparable, low CSPR inoculation suffered significantly in terms of reduced initial cell growth and impaired viability. This study underlines the importance to determine the CSPR for the design and implementation of an N-1 perfusion process in order to achieve the desired performance at the crucial production stage. Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.

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