Title

Microcarrier-based production of dengue virus in animal-free medium

Conference Dates

June 12-17, 2016

Abstract

Microcarriers have been a mainstay production platform for vaccine production for decades. They are used for production of both human and animal vaccines. However, many processes still use animal derived components in their manufacturing processes. The use of undefined components in animal product-free (APF) cell culture media is one source of variability in cell culture processes. Therefore a production medium optimized for use with microcarrier-based production systems would be desirable. In these studies, a hydrolysate-free and APF, Vero cell production medium that is ideal for use in stirred-tank vessels with Pall SoloHill® microcarriers was developed. During development, medium formulations were optimized through an iterative process in concert with various microcarrier types to support robust cell growth and virus production in small-scale spinners. Further optimization then occurred in larger bioreactors.

The medium supports sustained, high-density cell growth on multiple types of APF SoloHill® microcarriers over a seven day expansion cycle without need for medium exchange . Virus production has been demonstrated to be equal to or greater than DMEM containing FBS and two different commerically-available serum-free media. The peak of wild type dengue 2 virus production advances up to 3 days earler in microcarrier culture when compared to static conditions, and cumulative titer is increased.

Vero cells achieve cell densities of >3 M cells/mL without a medium exchange (batch culture) and maintain cell growth for 8-9 days. In mock infections, the medium enables cell densities of up to 6 M cells/mL in bioreactors.

Additional Files
Mark Szczypka - 61.pdf (6 kB)

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