Team Leader (USP)

PD Dr. Yvonne Genzel
PD Dr. Yvonne Genzel
Phone: +49 391 6110 257
Room: N0.18

Researcher

Felipe Tapia
Felipe Tapia
Phone: +49 391 6110 207
Room: N0.15

Additional Information

External collaborations:

ProBioGen AG, Berlin, Germany (Dr. Volker Sandig)

Internal collaborations: 

Pavel Marichal-Gallardo, BPE, Downstream Processing, MPI Magdeburg

Continuous Production of Vaccines in a Tubular Bioreactor System

Header image 1541420790

Continuous Production of Cell Culture-derived Vaccines in a Plug-flow Tubular Bioreactor System

Motivation

Using cascades of stirred tank bioreactors is a first approach for a continuous cell-culture based virus production. Another option for continuous virus production is the use of a plug-flow tubular bioreactor (PFBR) [1,2] within the cascade. As before, a chemostat is used for the continuous production of cells. However, in this set-up, virus seed is continuously added to the cells at the entry of a PFBR (Fig. 1). If the length of the tubing is adjusted to the MOI and the duration of the replication cycle, a virus harvest with defined passage number can be collected continuously at the tube outlet. The multiplicity of infection (MOI) at the point of entry is a function of the cell concentration in the chemostat, the virus concentration in the virus stock, and the flow rates, as described in Fig. 1. This has several advantages:

1) Elimination of the risk of viral antigenic variation as each cell that enters the tube is infected with a virus stock of defined passage number , and the number of additional virus passages inside the tube is limited

2) Steady-state operation allows harvesting of virus particles with defined quality attributes over extended time periods.

3) Suitable for production of viruses which show significant accumulation of defective interfering particles (DIPs) and display high mutation rates, i.e. influenza A virus [3,4,5]

Aim of the project

  • Design and optimization  of a PFBR system for production of influenza A virus (Fig. 2)
  • Characterization of the system by monitoring the total number of virus particles, the concentration of infectious virions, cells and metabolites using  flow cytometry, quantitative  PCR, and virus assays [3]
  • Establishment of a mathematical model  to describe process behavior
  • Process design and evaluation for other viruses and cells (optionsfor licensing and/or collaborations [1])

Fig. 1. Scheme of a continuous virus production process using a plug-flow tubular bioreactor (PFBR). Cells and viruses are mixed at the point of infection (POI) and move under a plug-flow regime along the tube. Virus propagation occurs within the volume of the tube and a harvest of defined virus passage number is produced over process time. Zoom Image

Fig. 1. Scheme of a continuous virus production process using a plug-flow tubular bioreactor (PFBR). Cells and viruses are mixed at the point of infection (POI) and move under a plug-flow regime along the tube. Virus propagation occurs within the volume of the tube and a harvest of defined virus passage number is produced over process time.

[less]
Fig. 2. Picture of the PFBR process (0.7 L total working volume) constructed at the MPI Magdeburg for continuous influenza A virus production at 0.20 mL/min with titers up to 2.5 log10 (HA units/100 µL) (6 L in 3 weeks) [1]. Scale-up is possible using one single tube of larger diameter or bundles of tubes. A system consisting of a 10 L chemostat connected to one PFBR, for example, would allow to collect 100 L of virus harvest within 19 days (3.7 mL/min). Zoom Image

Fig. 2. Picture of the PFBR process (0.7 L total working volume) constructed at the MPI Magdeburg for continuous influenza A virus production at 0.20 mL/min with titers up to 2.5 log10 (HA units/100 µL) (6 L in 3 weeks) [1]. Scale-up is possible using one single tube of larger diameter or bundles of tubes. A system consisting of a 10 L chemostat connected to one PFBR, for example, would allow to collect 100 L of virus harvest within 19 days (3.7 mL/min).

[less]
Fig. 3. Process diagram of the PFBR system (0.7 L total working volume) constructed at the MPI Magdeburg for continuous influenza A virus production at 0.20 mL/min with titers up to 2.5 log10 (HA units/100 µL) (6 L in 3 weeks) [1]. Scale-up is possible using one single tube of larger diameter or bundles of tubes. A system consisting of a 10 L chemostat connected to one PFBR, for example, would allow to collect 100 L of virus harvest within 19 days (3.7 mL/min). Zoom Image
Fig. 3. Process diagram of the PFBR system (0.7 L total working volume) constructed at the MPI Magdeburg for continuous influenza A virus production at 0.20 mL/min with titers up to 2.5 log10 (HA units/100 µL) (6 L in 3 weeks) [1]. Scale-up is possible using one single tube of larger diameter or bundles of tubes. A system consisting of a 10 L chemostat connected to one PFBR, for example, would allow to collect 100 L of virus harvest within 19 days (3.7 mL/min). [less]

References

Tapia, F., Genzel, Y., Reichl U.:
Plug flow bioreactor, method containing the same and method for virus production.
Levenspiel, O.:
Chemical reaction engineering.
Fogler, S.:
Elements of Chemical Reaction Engineering.
Frensing, T.; Heldt, S.; Pflugmacher, A.; Behrendt, I.; Jordan, I.; Flockerzi, D.; Genzel, Y.; Reichl, U.: Continuous Influenza Virus Production in Cell Culture Shows a Periodic Accumulation of Defective Interfering Particles. PLoS One 8 (9), p. e72288 (2013)
Hu, Y. C., Wang, M. Y., Bentley, W. E.:
A tubular segmented-flow bioreactor for the infection of insect cells with recombinant baculovirus.
Kompier et al.:
A continuous process for the production of baculovirus using insect-cell cultures.

 
loading content
Go to Editor View