Protocol

Baculovirus and Insect Cell Expression Protocols

Volume 388 of the series Methods in Molecular Biology™ pp 225-245

Recombinant Protein Production in Large-Scale Agitated Bioreactors Using the Baculovirus Expression Vector System

  • Cynthia B. EliasAffiliated withAnimal Cell Technology Group, Biotechnology Research Institute-National Research Council Canada
  • , Barbara JardinAffiliated withAnimal Cell Technology Group, Biotechnology Research Institute-National Research Council Canada
  • , Amine KamenAffiliated withAnimal Cell Technology Group, Biotechnology Research Institute-National Research Council Canada

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Abstract

The production of recombinant proteins using the baculovirus expression vector system in large-scale agitated bioreactors is discussed in this chapter. Detailed methods of the key stages of a batch process, including host cell growth, virus stock amplification and quantification, bioreactor preparation and operation, the infection process, final harvesting, and primary separation steps for recovery of the product are presented. Furthermore, methods involved with online monitoring and bioreactor control, which have a significant impact on the overall success of the process, are provided, including advanced online monitoring of physiological parameters such as biovolume and respiration activity for batch and fed-batch insect cell cultures along with their role in operating high cell density cultures.

Key Words

Bioreactor large-scale recombinant protein expression fed-batch high cell density insect cell lytic system baculovirus on-line monitoring serum free