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Microorganisms 2016, 4(2), 18; doi:10.3390/microorganisms4020018

Fed-Batch Production of Bacterial Ghosts Using Dielectric Spectroscopy for Dynamic Process Control

1
Research Center Pharmaceutical Engineering (RCPE) GmbH, Inffeldgasse 13, Graz A-8010, Austria
2
Research Division Biochemical Engineering, Institute of Chemical Engineering, Vienna University of Technology, Gumpendorfer Strasse 1A 166/4, Vienna A-1060, Austria
3
Biotech Innovation Research Development and Consulting (BIRD–C) GmbH & Co KG, Dr.-Bohr-Gasse 2-8, Vienna A-1030, Austria
4
Center of Molecular Biology, University of Vienna, Dr.-Bohr-Gasse 9, Vienna A-1030, Austria
*
Author to whom correspondence should be addressed.
Academic Editor: Willy Verstraete
Received: 3 February 2016 / Revised: 11 March 2016 / Accepted: 18 March 2016 / Published: 24 March 2016
View Full-Text   |   Download PDF [2120 KB, uploaded 24 March 2016]   |  

Abstract

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. View Full-Text
Keywords: E-lysis; bacterial ghosts; process analytical technology (PAT); online biomass monitoring; dielectric spectroscopy; control strategy E-lysis; bacterial ghosts; process analytical technology (PAT); online biomass monitoring; dielectric spectroscopy; control strategy
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MDPI and ACS Style

Meitz, A.; Sagmeister, P.; Lubitz, W.; Herwig, C.; Langemann, T. Fed-Batch Production of Bacterial Ghosts Using Dielectric Spectroscopy for Dynamic Process Control. Microorganisms 2016, 4, 18.

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