Optimisation of protein expression and establishment of the Wave Bioreactor for Baculovirus/insect cell culture |
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Authors: | Wilfried Weber Eric Weber Sabine Geisse Klaus Memmert |
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Affiliation: | (1) Core Technologies/Biomolecules Production, Novartis Pharma AG, Research, Building WSJ-506.101, CH-4002 Basel, Switzerland;(2) Present address: ETH Hoenggerberg, CH-8093 Zürich, Switzerland |
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Abstract: | As the interest of research is beginning to shift from genomicsto proteomics the number of proteins to be expressed is rapidlyincreasing. To do so, well-established, high-level expressionsystems and rapid, cost-effective production means are needed. For addressing the latter, a novel cultivation system for recombinant cells, the Wave Bioreactor has recently becomeavailable. We describe the set-up and the optimisation of parameters essential for successful operation and growth of insect cells to high cell densities in the Wave Bioreactor. According to our experience, the Cellbag system comparesvery favorably to conventional cultivation vessels such as bioreactors and roller cultures with respect to simplicity ofoperation and cost. Additionally, we developed a rapid and simple protocol for assessing expression and production conditions for the Baculovirus/insect cell system applicable to many different genes/proteins. Important parameters like MOI,TOI, peak cell density (PCD) and expression levels are determinedin pre-experiments on small scale to achieve optimal expressionof a given protein. These conditions are subsequently transformedand applied to large scale cultures grown in nutrient-supplemented medium in the Wave Bioreactor. |
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Keywords: | baculovirus/insect cell system essential parameters for operation expression optimisation feeding strategies Wave Bioreactor |
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