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

Abstract

As the interest of research is beginning to shift from genomics to proteomics the number of proteins to be expressed is rapidly increasing. To do so, well-established, high-level expression systems and rapid, cost-effective production means are needed. For addressing the latter, a novel cultivation system for recombinant cells, the Wave Bioreactor™ has recently become available. 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 compares very favorably to conventional cultivation vessels such as bioreactors and roller cultures with respect to simplicity of operation 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 determined in pre-experiments on small scale to achieve optimal expression of a given protein. These conditions are subsequently transformed and applied to large scale cultures grown in nutrient-supplemented medium in the Wave Bioreactor.