Applied Microbiology and Biotechnology

, Volume 102, Issue 24, pp 10469–10483 | Cite as

A new strategy for fed-batch process control of HEK293 cell cultures based on alkali buffer addition monitoring: comparison with O.U.R. dynamic method

  • I. Martínez-MongeEmail author
  • P. Comas
  • J. Triquell
  • M. Lecina
  • A. Casablancas
  • J. J. Cairó
Biotechnological products and process engineering


The increasing demand for biopharmaceuticals produced in mammalian cells has driven the industry to enhance productivity of bioprocesses through different strategies. This is why fed-batch and perfusion cultures are considered more attractive choices than batch processes. In this context, the availability of reliable online measuring systems for cell density and metabolic activity estimation will help the application of these processes. The present work focuses on the comparison of two different monitoring tools for indirect estimation of biomass concentration in a HEK293 cell cultures producing IFN-γ: on one side, the oxygen uptake rate (O.U.R.) determination, by means of application of the dynamic method measurement which is already a widely used tool and, on the other side, a new robust online monitoring tool based on the alkali buffer addition used to maintain the pH set point. Both strategies allow a proper monitoring of cell growth and metabolic activity, with precise identification of the balanced cell growth and the most important action in the process, as is the media feeding. The application of these monitoring systems in fed-batch processes allows extending the growth of HEK293 cells, which in turn results in higher final cell concentrations compared with Batch strategy (7 · 106 cells mL−1), achieving 14 · 106 cells mL−1 for the fed-batch based on O.U.R. and 19 · 106 cells mL−1 for the fed-batch based on the alkali addition. Product titter is also increased in respect of the batch strategy (3.70 mg L−1), resulting in 8.27 mg L−1 when fed-batch was based on O.U.R. and 11.49 mg L−1 when it was based on the alkali buffer strategy. Results prove that fed-batch strategy based on the alkali buffer addition is a robust online monitoring method that has shown its great potential to optimize the feeding strategy in HEK293 fed-batch cultures.


HEK293 O.U.R. Cell culture Fed-batch Lactate Alkali buffer addition monitoring 



The present work has been supported by the grant FI-DGR (Generalitat de Catalunya, Catalonia, Spain).

Compliance with ethical standards

Ethical statements

The authors are aware with the ethical responsibilities required by the journal and are committed to comply them.

Conflict of interest

The authors declare that they have no competing interests.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2018_9388_MOESM1_ESM.pdf (218 kb)
ESM 1 (PDF 218 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Chemical, Biological and Environmental EngineeringUniversitat Autònoma de BarcelonaCerdanyola del VallèsSpain
  2. 2.IQS School of EngineeringUniversitat Ramon LlullBarcelonaSpain

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