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. 2002 Jan;38(1-3):77–85. doi: 10.1023/A:1021102015070

Optimisation of protein expression and establishment of the Wave Bioreactor for Baculovirus/insect cell culture

Wilfried Weber 1, Eric Weber 1, Sabine Geisse 1, Klaus Memmert 1,✉
PMCID: PMC3449929  PMID: 19003089

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.

Keywords: baculovirus/insect cell system, essential parameters for operation, expression optimisation, feeding strategies, Wave Bioreactor

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