Fed-Batch Production of Bacterial Ghosts Using Dielectric Spectroscopy for Dynamic Process Control.
The Bacterial Ghost (BG) platform technology evolved from a microbiological expression system incorporating the ϕX174 lysis gene E. E-lysis generates empty but structurally intact cell envelopes (BGs) from Gram-negative bacteria which have been suggested as candidate vaccines, immunotherapeutic agents or drug delivery vehicles. E-lysis is a highly dynamic and complex biological process that puts exceptional demands towards process understanding and control. The development of a both economic and robust fed-batch production process for BGs required a toolset capable of dealing with rapidly changing concentrations of viable biomass during the E-lysis phase. This challenge was addressed using a transfer function combining dielectric spectroscopy and soft-sensor based biomass estimation for monitoring the rapid decline of viable biomass during the E-lysis phase. The transfer function was implemented to a feed-controller, which followed the permittivity signal closely and was capable of maintaining a constant specific substrate uptake rate during lysis phase. With the described toolset, we were able to increase the yield of BG production processes by a factor of 8-10 when compared to currently used batch procedures reaching lysis efficiencies >98%. This provides elevated potentials for commercial application of the Bacterial Ghost platform technology.
Research Center Pharmaceutical Engineering (RCPE) GmbH, Inffeldgasse 13, Graz A-8010, Austria. andrea.meitz@tuwien.ac.at.
Andrea
A
Meitz
Research Division Biochemical Engineering, Institute of Chemical Engineering, Vienna University of Technology, Gumpendorfer Strasse 1A 166/4, Vienna A-1060, Austria. patrick.sagmeister@exputec.com.
Patrick
P
Sagmeister
Biotech Innovation Research Development and Consulting (BIRD-C) GmbH & Co KG, Dr.-Bohr-Gasse 2-8, Vienna A-1030, Austria. werner.lubitz@bird-c.at.|Center of Molecular Biology, University of Vienna, Dr.-Bohr-Gasse 9, Vienna A-1030, Austria. werner.lubitz@bird-c.at.
Werner
W
Lubitz
Research Division Biochemical Engineering, Institute of Chemical Engineering, Vienna University of Technology, Gumpendorfer Strasse 1A 166/4, Vienna A-1060, Austria. christoph.herwig@tuwien.ac.at.
Christoph
C
Herwig
Research Center Pharmaceutical Engineering (RCPE) GmbH, Inffeldgasse 13, Graz A-8010, Austria. mail@timo-langemann.de.|Biotech Innovation Research Development and Consulting (BIRD-C) GmbH & Co KG, Dr.-Bohr-Gasse 2-8, Vienna A-1030, Austria. mail@timo-langemann.de.
Timo
T
Langemann
E-lysis; bacterial ghosts; control strategy; dielectric spectroscopy; online biomass monitoring; process analytical technology (PAT)
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