Researcher

Susann Freund
Phone:+49 391 6110 228

Sandtorstrasse 1, 39106 Magdeburg, Germany

Alexander Rath
Email:rath@...

Additional Information

Collaborations:

• within SysLogics project:
→ Prof. Dr. Thomas Noll, Institute of Cell Culture Technology (University of Bielefeld).
→ Prof. Dr. rer. nat. An-Ping Zeng, Institute of Bioprocess and Biosystems Engineering (University of Hamburg-Harburg).
→ Prof. Dipl. Ing. Dr. tech. Elmar Heinzle, Research Group Biochemical Engineering (University of Saarbruecken).
→ ProBioGen AG
→ (Funded by BMBF-initiative FORSYS)

• Dr. Steffen Klamt, Max Planck Institute for Dynamics of Complex Technical Systems (MPI Magdeburg)

• Prof. Dr.-Ing Rolf Findeisen, Institute for Automation Engineering - IFAT, Otto-von-Guericke University Magdeburg

Start:
2008/07/03

End: April 2013

SysLogics - Modelling of Central Metabolism

SysLogics - Modelling of Central Metabolism

Motivation

Since many years mammalian cells are cultivated to produce monoclonal antibodies, recombinant proteins or vaccines. New designer cell lines as alternatives to well-established cell lines are now under consideration for production of these biologics. These cell lines are fully documented, meet regulatory requirements and are selected for a particular production process. In the SysLogics project the novel human designer cell lines AGE1.HN and its clone AGE1.HN.AAT are use to study their metabolism, proteome and genome [1,2]. In a first attempt process characterization is performed by determination of cell numbers, product formation, uptake of substrates and release of metabolites. Subsequent several process control strategies can be applied to increase productivity and reproducibility of the production process. In addition, development of mathematical models could help in experimental design, can be used for process control and may provide better understanding of the dynamics of central metabolism. Here a novel human neuronal designer cell line supplied by an industrial partner (ProBioGen AG) is cultivated under varied physiological conditions in batch and continuous cultivation mode.

Work strategy

  • Batch cultivations for determination of kinetics and parameters for calculation of metabolic rates in the pseudo steady state of the exponential growth phase as well as comparison of batch experiment, collaboration with SysLogics partners
  • model formation based on reliable determination of growth influencing factors in collaboration with Prof. Dr.-Ing. Rolf Findeisen

  • Continuous cultivations under substrate limitation

  • Continuous cultivations with different media compositions

  • analysis of steady states of continuous cultivations with metabolic flux analysis, therefore comparison of the producing cell line AGE1.HN with non producing cell line AGE1.HN.AAT in collaboration with Dr. Steffen Klamt
Figure 1: Scheme of continuous cultivation experiments Zoom Image
Figure 1: Scheme of continuous cultivation experiments

Modelling of a novel human designer cell line

The modelling and especially the metabolic flux analysis of a cell line requires media with defined composition. Ideally, for mammalian cell culture the medium should neither contain serum nor lysates. With data from batch and continuous cultivations under various physiological conditions several modelling approaches will be pursued.

Research aims

  • Creation of suitable structured, unsegregated model
  • Analysis of metabolic pathways by metabolic flux analysis based on a model for MDCK cells with 112 reactions [3,4]
Figure 2: Simulation of continuous cultivation experiments Zoom Image
Figure 2: Simulation of continuous cultivation experiments
Figure 3: Metabolic fluxes of a MDCK cell Zoom Image
Figure 3: Metabolic fluxes of a MDCK cell


References

[1] Niklas J.; Schrader E.; Sandig V.; Noll T.; Heinzle E., "Quantitative characterization of metabolism and metabolic shifts during growth of the new human cell line AGE1.HN using time resolved" Bioprocess Biosyst Eng, 2011, (34) 533-545.
[2] Barradas P.O.; Pörtner, R., Jandt U, Phan LDM, Villanueva ME, Rath A, Reichl U, Schräder, E., Scholz S, Noll T et al: Evaluation of criteria for bioreactor comparison and operation standardisation for mammalian cell culture. Engineering in Life Sciences 2012.
[3] Moehler, L.; Bock, A. & Reichl, U. "Segregated mathematical model for growth of anchorage-dependent MDCK cells in microcarrier culture." Biotechnology Progress, 2008, 24(1): 110-119.
[4] Wahl, A.; Sidorenko, Y.; Dauner, M.; Genzel, Y. & Reichl, U. "Metabolic flux model for an anchorage-dependent MDCK cell line: characteristic growth phases and minimum substrate consumption flux distribution." Biotechnology and Bioengineering, 2008, 101(1): 135-152.

Related projects

Process Characterisation and Optimisation for a Recombinant Protein Produced by a Novel Human Designer Cell Line
Quantitative Analysis of Energy Metabolism of Animal Cells
Enzymatic Characterization of Mammalian Cells

 
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