Biotechnological aspects of the production of human pro-kallikrein using the AcNPV-baculovirus-expression system |
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| Authors: | Georg Fertig Hans Peter Rahn Axel Angermann Martina Klöppinger Herbert Georg Miltenburger |
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| Institution: | (1) Institute of Zoology, Cell Biology Laboratory, Technical University of Darmstadt, Petersenstr. 22, D-6100 Darmstadt, Germany;(2) Institute of Biochemistry, Technical University of Darmstadt, Petersenstr. 22, D-6100 Darmstadt, Germany;(3) Institute for Peptide Research (IPF) GmbH, Feodor Lynen Str. 61, D-3000 Hannover, Germany |
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| Abstract: | We have investigated large scale production processes (up to 2 liters) of recombinant proteins using the baculovirus expression system in order to optimize the product yields. Experiments using cell lines ofSpodoptera frugiperda (Sf9) andMamestra brassicae (IZD-Mb0503) were performed to show the different production capacities of the cell lines. The influence of the infection at different cell densities is described. Beyond that, TC100-, IPL41- and serum-free IPL41-medium were compared to demonstrate their different capabilities of supporting cell growth and protein expression. Additionally, the inhibitory effect of FCS on the protease activity of kallikrein, which is produced in its zymogenic form, is discussed. Improved production parameters are described, which enabled us to produce up to 8000 units of activated pro-kallikrein within 14 days using perfusion cultivation.Abbreviations AcNPV
autographa californica nuclear polyhedrosis virus
- MOI
multiplicity of infection |
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| Keywords: | baculovirus-expression system human pro-kallikrein batch- and perfusion fermentation media comparison process development |
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