Development of an animal-component free insect medium for the Baculovirus Expression Vector System (BEVS)

Conference Dates

June 17-22, 2018


Insect cells derived from Spodoptera frugiperda have been widely used with the baculovirus expression vector system (BEVS) for the production of recombinant proteins and adeno-associated viruses (AAVs) due to their ease of culture, scalability in high cell density suspension cultures, and high protein expression levels. Traditionally, insect cells are cultured in an undefined medium containing yeast hydrolysate and cod liver oil, however, there is an increasing push to use chemically defined, animal-component free medium to minimize any potential contaminants and decrease lot-to-lot variability while maintaining high cell growth and production. In this case study, an animal-component free insect medium was developed utilizing Rational Culture Media DesignTM and evaluated with Sf9 cells. Using a traditional formulation as a starting point, the final medium was developed by optimizing multiple nutrient groups in the basal medium, replacing the animal-derived components, and screening several yeast hydrolysate sources. By utilizing multifactor design of experiment software, various nutrient groups were screened including amino acids, vitamins, and metals. The metals group was identified to have the most impact on cell growth and productivity, and therefore concentrations of metal components were further optimized. In addition, the animal-derived components in the starting formulation, cod liver oil and cholesterol, were replaced with animal-component free fatty acids and synthetic cholesterol, respectively. The concentrations of these components were optimized to achieve better growth performance and production while also sustaining formulation stability and streamlining manufacturing processes. Finally, yeast hydrolysate is a well-known, undefined component that is crucial for insect cell growth and productivity. To minimize lot-to-lot variability, the yeast hydrolysate concentration was significantly lowered, and multiple yeast hydrolysate sources and lots were evaluated to determine the highest quality source. As a result, an animal-component free insect medium was developed that had improved growth performance and comparable productivity to a widely used commercially available animal-derived medium.

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