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Improving bioreactor cultivation conditions for sensitive cell lines by dynamic membrane aeration

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Abstract

Although the importance of animal cell culture for the industrial (large scale) production of pharmaceutical products is continuously increasing, the sensibility of the cells towards their cultivation environment is still a challenging issue. In comparison to microbial cultures, cell cultures which are not protected by a cell wall are much more sensitive to shear stress and foam formation. Reactor design as well as the selection of ‘robust’ cell lines is particularly important for these circumstances. Nevertheless, even ‘sensitive’ cell lines are selected for certain pharmaceutical processes due to various reasons. These sensitive cell lines have even higher requirements regarding their cultivation environment. Important characteristics for the corresponding reactor design are a high (volumetric) gas mass transfer coefficient, low volumetric power input, low shear stress, low susceptibility to bio-fouling, the ability to cultivate sticky cells and sufficient mixing properties. Membrane aeration has been a long-known possibility to meet some of these requirements, but has not often been applied in recent years. The reasons lie mainly in low gas mass transfer rates, a limited installable volume-specific membrane surface area, restrictions in scalability and problems with membrane fouling. The dynamic membrane aeration bioreactor aeration is a simple concept for bubble-free oxygen supply of such sensitive cultures. It overcomes limitations and draw-backs of previous systems. Consisting of an oscillating, centrally arranged rotor (stirrer) that is wrapped with silicone membrane tubing, it enables doubling the gas mass transfer at the same shear stress in the investigated cultivation scales of 12, 20, 100, and 200 L. Continuous cultivation at these scales allows the same product output as fed-batch cultivation does at tremendously larger reactor volumes. Apart from introducing this novel technology, the presentation comprises selected cultivation results obtained for blood coagulation factor VIII in continuous mode and a therapeutic monoclonal antibody in fed-batch mode in comparison to reference trials.

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Abbreviations

a :

Fermenter liquid volume-specific membrane surface area (m2 m−3)

d ef :

Equivalent floc diameter (μm)

k :

(Oxygen) mass transfer coefficient (m h−1)

ka :

Volume-specific (oxygen) mass transfer coefficient (h−1)

P :

Power input (W)

P/V :

Volume-specific power input (W m−3)

V :

Fermenter (liquid) volume (m3)

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Acknowledgments

This development of the DMA was funded by Bayer Technology Services (BTS) within two consecutive research projects. We would like to thank to Messrs. Rose, Gießelmann, and Grodotzki for carrying out the reactor design drawings and to colleagues form Tectrion for manufacturing the reactors (Messrs Commer, Schiffczyk, Drinhausen, Kamphusmann, and Letzner). We wish to thank our colleagues in the Enzyme & Fermentation Technology group of BTS for their support. We express our gratitude to the students Andreas Sinthern, Juliane Schröter, and Maike Rampe for their contributions made in the context of their diploma theses. The described fed-batch cell cultivations with the 20 and 200 L bioreactors took place on the premises of Bayer Health Care AG in Wuppertal. For this, special thanks are due to Mr. Wischniewski and his colleagues. The continuous cell cultivations with the 12 L bioreactors were performed at Bayer Health Care in Berkeley, USA. Special thanks for this to Klaus Jöris and his colleagues.

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Authors and Affiliations

  1. Bayer Technology Services (BTS), Enzyme and Fermentation Technology, 51368, Leverkusen, Germany

    Björn Frahm & Helmut Brod

  2. Bayer HealthCare AG, Global Biological Development, 42096, Wuppertal, Germany

    Uwe Langer

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  1. Björn Frahm
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  2. Helmut Brod
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  3. Uwe Langer
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Correspondence to Björn Frahm.

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Frahm, B., Brod, H. & Langer, U. Improving bioreactor cultivation conditions for sensitive cell lines by dynamic membrane aeration. Cytotechnology 59, 17–30 (2009). https://doi.org/10.1007/s10616-009-9189-9

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  • DOI: https://doi.org/10.1007/s10616-009-9189-9

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