Downstream Processing

Downstream Processing


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 (bio)pharmaceuticals. This is mainly due to the high demands on purity, removal of contaminants, and product safety. In order to develop efficient downstream processes it is not only necessary to improve existing purification methods and to introduce new unit operations but also to reconsider complete downstream processing trains.

Currently, our research group focuses on the development of novel platform technologies for the purification of virus particles (influenza virus, vaccinia virus, flaviviruses), viral vectors, virus like particles and pharmaceutically relevant glycoproteins (e.g. 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.

Fig.1. Current downstream processing activities at BPE

Example Influenza Vaccines

Our work on downstream processing of influenza virus aims at the exploration and development of a purification process for cell culture-derived vaccines. Unit operations like ultrafiltration, size-exclusion, ion-exchange, affinity, steric exclusion, 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.

Fig. 2 Options for downstream processing in influenza vaccine manufacturing


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Hämmerling, F.; Pieler, M.; Hennig, R.; Serve, A.; Rapp, E.; Wolff, M. W.; Reichl, U.; Hubbuch, J.: Influence of the production system on the surface properties of influenza A virus particles. Engineering in Life Sciences 17 (10), pp. 1071 - 1077 (2017)
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Weigel, T.; Solomaier, T.; Wehmeyer, S.; Peuker, A.; Wolff, M. W.; Reichl, U.: A membrane-based purification process for cell culture-derived influenza A virus. Journal of Biotechnology 220, pp. 12 - 20 (2016)
Wolff, M. W., Pieler, M. M., Marichal-Gallardo, P., Reichl, U.
Method for the separation of virus compositions including depletion and purification thereof.
EU Patent: WO 2017/076553 A1 (2016)
Serve, A.; Pieler, M.; Benndorf, D.; Rapp, E.; Wolff, M. W.; Reichl, U.: Comparison of Influenza Virus Particle Purification Using Magnetic Sulfated Cellulose Particles with an Established Centrifugation Method for Analytics. Analytical Chemistry 87 (21), pp. 10708 - 10711 (2015)
Weigel, T.; Solomaier, T.; Peuker, A.; Pathapati, T.; Wolff, M. W.; Reichl, U.: A flow-through chromatography process for influenza A and B virus purification. Journal of Virological Methods 207, pp. 45 - 53 (2014)
Kröber, T.; Wolff, M. W.; Hundt, B.; Seidel-Morgenstern, A.; Reichl, U.: Continuous purification of influenza virus using simulated moving bed chromatography. Journal of Chromatography A 1307, pp. 99 - 110 (2013)
Wolff, M. W., Opitz, L., Reichl, U.
Method for the preparation of sulfated cellulose mebranes and sulfated cellulose membranes.
EU Patent: EP 2144937 B1 (2012)
US Patent: US 8,173,021 B2 (2012)
Post-Hansen, S., Faber, R., Reichl, U., Wolff, M. W., Gram, A. P.
Purification of Vaccinia viruses using hydrophobic interaction chromatography.
US Patent: US 8,470,578 B2 (2010)
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)
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)

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