Team Leader (DSP)

Prof. Dr. Michael Wolff
Prof. Dr. Michael Wolff

Profile

Universitätsplatz 2, Gebäude 25, 39106 Magdeburg, Germany

Team (DSP)

Pavel Marichal-Gallardo, M. Sc.
Pavel Marichal-Gallardo, M. Sc.
Phone: +49 391 67 546 79
Fax: +49 391 6110 500
Room: G25-118
Anja Bastian (parental leave)
Technical Assistant
Phone: +49 391 67 546 70
Room: G25-123
Annika Deinert
Technical Assistant
Phone: +49 391 67 546 69
Room: G25-123

Additional Information

Collaborations with industry:

  • BIA Separations Inc.
  • EMC microcollections GmbH
  • IDT Biologika GmbH
  • Merckle Biotec GmbH
  • Novartis Vaccines and Diagnostics GmbH & Co. KG
  • Sartorius Stedim Biotech GmbH
  • Sentinext Therapeutics Sdn Bhd
  • Tosoh Bioscience LLC

Collaborations with academia:

  • Federal University of Rio de Janeiro, Rio de Janeiro, Brazil; Chemical Engineering Program Cell Culture Engineering Laboratory (Prof. Dr. Leda dos Reis Castilho)
  • Karlsruhe Institute of Technology, Karlsruhe; Biomolecular Separation Engineering (Prof. Dr. Jürgen Hubbuch)
  • Max Planck Institute for Dynamics of Complex Technical Systems, Magdeburg, Germany; Physical and Chemical Foundations of Process Engineering Group (Prof. Dr.-Ing. Andreas Seidel-Morgenstern)
  • Max Planck Institute for Dynamics of Complex Technical Systems, Magdeburg, Germany; Physical and Chemical Process Engineering Group (Prof. Dr.-Ing. Kai Sundmacher)

Downstream Processing

Header image 1377693701

Downstream Processing

Motivation

The production of recombinant proteins and vaccines is a rapidly growing field in biotechnological industry. Manufacturing of biologicals is a complex task ranging from strain development and upstream processing to the purification and formulation of the product. At the beginning of the biotechnology era product concentrations in bioreactors were within the mg per liter range. Currently, yields of up to 10 g per liter of fermenter harvest are obtained in monoclonal antibody production. These achievements have been mainly accomplished by the use of high expression cell lines, media optimization, and an increase in cell numbers using appropriate cultivation conditions. However, while bioreactor yields were improved significantly, optimization of downstream processing was often neglected and now constitutes a bottleneck in various manufacturing processes.

Downstream processing often accounts for the major part of production costs of pharmaceuticals. This is mainly due to the high demands on purity, removal of contaminants and product safety. In order to develop efficient processes it is not only necessary to improve existing purification methods and to introduce new unit operations but also to design complete downstream processing trains.

Currently, our research group focuses on the development of novel applications and techniques for the purification of virus particles (influenza virus, Vaccinia virus), virus like particles as well as downstream processing of pharmaceutically relevant glycoproteins (erytropoetin, factor VIII). Furthermore we investigate the aggregation behavior of macromolecular biological components and virus particles. An overview of the different activities is given below.

<em>Fig.1. Current downstream processing activities at BPE.</em> Zoom Image
Fig.1. Current downstream processing activities at BPE.

Influenza Vaccines

Our work on downstream processing of influenza virus aims at the exploration and development of a purification process for cell culture-derived inactivated whole virus vaccines. Unit operations like ultrafiltration, size-exclusion, ion-exchange, affinity, and hydrophobic interaction chromatography are combined to an overall process train. To improve performance and productivity of the DSP we focus on the use of modern resins, membrane adsorbers and monoliths as well as continuous methods like simulated moving bed chromatography.

<em>Fig.2. Options for downstream processing in influenza vaccine manufacturing </em> Zoom Image
Fig.2. Options for downstream processing in influenza vaccine manufacturing

Smallpox vaccines (Michael Wolff; OvGU Collaboration)

The current downstream process project for smallpox vaccines aims at the development of purification schemes for chicken embryo fibroblast cell-culture derived Modified Vaccinia Ankara virus (MVA-BN®). The process trains are based on a combination of pseudo-affinity MA, ion-exchange MA, HIC, and diafiltration steps. Pseudo-affinity MA are based on sulfated carbohydrates like heparin and cellulose-sulfate, whereas sulfated cellulose MA are developed and produced in our laboratories.

<em>Fig.3. Options for downstream processing in smallpox vaccine manufacturing </em> Zoom Image
Fig.3. Options for downstream processing in smallpox vaccine manufacturing

References

1.
Wolff, M.; Reichl, U.: Downstream processing of cell culture-derived virus particles. Expert Review of Vaccines 10 (10), pp. 1451 - 1475 (2011)
2.
Wolff, M. W., Reichl, U., Opitz, L.
Preparation of sulfated cellulose membranes for isolation of viruses and proteins.
3.
Wolff, M.; Siewert, C.; Hansen, S. P.; Faber, R.; Reichl, U.: Purification of cell culture-derived modified Vaccinia Ankara virus by pseudo-affinity membrane adsorbers and hydrophobic interaction chromatography. Biotechnology and Bioengineering 107 (2), pp. 312 - 320 (2010)
4.
Wolff, M.; Siewert, C.; Lehmann, S.; Hansen, S.P.; Djurup, R.; Faber, R.; Reichl, U.: Capturing of Cell Culture-Derived Modified Vaccinia Ankara Virus by Ion Exchange and Pseudo-Affinity Membrane Adsorbers. Biotechnology and Bioengineering 105 (4), pp. 761 - 769 (2010)
5.
Post-Hansen, S., Faber, R., Reichl, U., Wolff, M.W., Gram, A.P.
Purification of vaccinia viruses using hydrophobic interaction chromatography.
6.
Kröber, T.; Knöchlein, A.; Eisold, K.; Kalbfuß-Zimmermann, B.; Reichl, U.: DNA Depletion by Precipitation in the Purification of Cell Culture-Derived Influenza Vaccines. Chemical Engineering and Technology 33 (6), pp. 941 - 959 (2010)
7.
Opitz, L.; Hohlweg, J.; Reichl, U.; Wolff, M. W.: Purification of cell culture-derived influenza virus A/Puerto Rico/8/34 by membrane-based immobilized metal affinity chromatography. Journal of Virological Methods 161 (2), pp. 312 - 316 (2009)
8.
Opitz, L.; Lehmann, S.; Reichl, U.; Wolff, M. W.: Sulfated membrane adsorbers for economic pseudo-affinity capture of influenza virus particles. Biotechnology and Bioengineering 103 (6), pp. 1144 - 1154 (2009)
9.
Opitz, L.; Zimmermann, A.; Lehmann, S.; Genzel, Y.; Lübben, H.; Reichl, U.; Wolff, M. W.: Capture of cell culture-derived influenza virus by lectins: strain independent, but host cell dependent. Journal of Virological Methods 154 (1-2), pp. 61 - 68 (2008)
10.
Kalbfuss, B.; Knöchlein, A.; Kröber, T.; Reichl, U.: Monitoring Influenza Virus Content in Vaccine Production: Precise Assays for the Quantitation of Hemagglutination and Neuraminidase Activity. Biologicals 36 (3), pp. 145 - 161 (2008)
11.
Kalbfuss, B.; Flockerzi, D.; Seidel-Morgenstern, A.; Reichl, U.: Size-exclusion chromatography as a linear transfer system: Purification of human influenza virus as an example. Journal of Chromatography B 873 (1), pp. 102 - 112 (2008)



 
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