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A model-based framework for parallel scale-down fed-batch cultivations in mini-bioreactors for accelerated phenotyping.

Abstract

Concentration gradients that occur in large industrial-scale bioreactors due to mass transfer limitations have significant effects on process efficiency. Hence, it is desirable to investigate the response of strains to such heterogeneities to reduce the risk of failure during process scale-up. Although there are various scale-down techniques to study these effects, scale-down strategies are rarely applied in the early developmental phases of a bioprocess, as they have not yet been implemented on small-scale parallel cultivation devices. In this study, we combine mechanistic growth models with a parallel mini-bioreactor system to create a high-throughput platform for studying the response of Escherichia coli strains to concentration gradients. As a scaled-down approach, a model-based glucose pulse feeding scheme is applied and compared with a continuous feed profile to study the influence of glucose and dissolved oxygen gradients on both cell physiology and incorporation of noncanonical amino acids into recombinant proinsulin. The results show a significant increase in the incorporation of the noncanonical amino acid norvaline in the soluble intracellular extract and in the recombinant product in cultures with glucose/oxygen oscillations. Interestingly, the amount of norvaline depends on the pulse frequency and is negligible with continuous feeding, confirming observations from large-scale cultivations. Most importantly, the results also show that a larger number of the model parameters are significantly affected by the scale-down scheme, compared with the reference cultivations. In this example, it was possible to describe the effects of oscillations in a single parallel experiment. The platform offers the opportunity to combine strain screening with scale-down studies to select the most robust strains for bioprocess scale-up.

Authors+Show Affiliations

Department of Bioprocess Engineering, Institute of Biotechnology, Technische Universität Berlin, Berlin, Germany.Biologics Development: Microbial Dev't, Sanofi-Aventis Deutschland GmbH, Frankfurt, Germany.Department of Bioprocess Engineering, Institute of Biotechnology, Technische Universität Berlin, Berlin, Germany.Department of Bioprocess Engineering, Institute of Biotechnology, Technische Universität Berlin, Berlin, Germany.Department of Bioprocess Engineering, Institute of Biotechnology, Technische Universität Berlin, Berlin, Germany.Biologics Development: Microbial Dev't, Sanofi-Aventis Deutschland GmbH, Frankfurt, Germany.Biologics Development: Microbial Dev't, Sanofi-Aventis Deutschland GmbH, Frankfurt, Germany.Department of Bioprocess Engineering, Institute of Biotechnology, Technische Universität Berlin, Berlin, Germany.Department of Bioprocess Engineering, Institute of Biotechnology, Technische Universität Berlin, Berlin, Germany. Department of Chemistry and Applied Biosciences, ETH Zurich-Institute of Chemical and Bioengineering, Zurich, Switzerland. DataHow AG, Zurich, Switzerland.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31317526

Citation

Anane, Emmanuel, et al. "A Model-based Framework for Parallel Scale-down Fed-batch Cultivations in Mini-bioreactors for Accelerated Phenotyping." Biotechnology and Bioengineering, 2019.
Anane E, García ÁC, Haby B, et al. A model-based framework for parallel scale-down fed-batch cultivations in mini-bioreactors for accelerated phenotyping. Biotechnol Bioeng. 2019.
Anane, E., García, Á. C., Haby, B., Hans, S., Krausch, N., Krewinkel, M., ... Cruz Bournazou, M. N. (2019). A model-based framework for parallel scale-down fed-batch cultivations in mini-bioreactors for accelerated phenotyping. Biotechnology and Bioengineering, doi:10.1002/bit.27116.
Anane E, et al. A Model-based Framework for Parallel Scale-down Fed-batch Cultivations in Mini-bioreactors for Accelerated Phenotyping. Biotechnol Bioeng. 2019 Jul 17; PubMed PMID: 31317526.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A model-based framework for parallel scale-down fed-batch cultivations in mini-bioreactors for accelerated phenotyping. AU - Anane,Emmanuel, AU - García,Ángel Córcoles, AU - Haby,Benjamin, AU - Hans,Sebastian, AU - Krausch,Niels, AU - Krewinkel,Manuel, AU - Hauptmann,Peter, AU - Neubauer,Peter, AU - Cruz Bournazou,Mariano Nicolas, Y1 - 2019/07/17/ PY - 2019/04/12/received PY - 2019/06/25/revised PY - 2019/07/09/accepted PY - 2019/7/19/pubmed PY - 2019/7/19/medline PY - 2019/7/19/entrez KW - Escherichia coli KW - mini-bioreactors KW - modeling KW - scale-down KW - scale-up effects JF - Biotechnology and bioengineering JO - Biotechnol. Bioeng. N2 - Concentration gradients that occur in large industrial-scale bioreactors due to mass transfer limitations have significant effects on process efficiency. Hence, it is desirable to investigate the response of strains to such heterogeneities to reduce the risk of failure during process scale-up. Although there are various scale-down techniques to study these effects, scale-down strategies are rarely applied in the early developmental phases of a bioprocess, as they have not yet been implemented on small-scale parallel cultivation devices. In this study, we combine mechanistic growth models with a parallel mini-bioreactor system to create a high-throughput platform for studying the response of Escherichia coli strains to concentration gradients. As a scaled-down approach, a model-based glucose pulse feeding scheme is applied and compared with a continuous feed profile to study the influence of glucose and dissolved oxygen gradients on both cell physiology and incorporation of noncanonical amino acids into recombinant proinsulin. The results show a significant increase in the incorporation of the noncanonical amino acid norvaline in the soluble intracellular extract and in the recombinant product in cultures with glucose/oxygen oscillations. Interestingly, the amount of norvaline depends on the pulse frequency and is negligible with continuous feeding, confirming observations from large-scale cultivations. Most importantly, the results also show that a larger number of the model parameters are significantly affected by the scale-down scheme, compared with the reference cultivations. In this example, it was possible to describe the effects of oscillations in a single parallel experiment. The platform offers the opportunity to combine strain screening with scale-down studies to select the most robust strains for bioprocess scale-up. SN - 1097-0290 UR - https://www.unboundmedicine.com/medline/citation/31317526/Model-based_framework_for_parallel_scale_down_fed-batch_cultivations_in_mini-bioreactors_for_accelerated_phenotyping L2 - https://doi.org/10.1002/bit.27116 DB - PRIME DP - Unbound Medicine ER -