Development of a High Efficiency Baculovirus/Mammalian Cell Expression System for Recombinant Protein mGM-CSF Production

Recently, many systems have been developed for the production of protein with medical significance. The baculovirus expression vector system is one of the best expression systems for high yield protein production in insect and mammalian cells with proper post-translational modifications. The baculovirus/insect cell expression vector system (BEVS) is one of the therapy protein expression systems in eukaryotic expression at present. The system had demonstrate outstanding performance in the foreign gene expression、gene therapy and application on bio-insecticide. However, the cellular lytic property of bcaulovirus/ insect cell system influence the quality of the heterologous protein produced and discontinue the fermentation process. The baculovirus/mammalian cell expression vector system could produce high quality heterologous protein need not post transformation. However, high multiplicity of infection (MOI) is necessary to increase productivity. In this research we will develop one baculovirus/mammalian cell bioreactor to solve the problem as: (1) the discontinue of baculovirus’s insect cell host to product heterologous protein because of the cellular lytic property in bcaulovirus/insect cell system, (2) the poor heterologous protein quality produced in bcaulovirus/insect system because of its cellular lytic property, and (3) the high MOI needed in mammalian cell to increase productivity of heterologous protein. We will setup one new baculovirus/mammalian cell expression vector system by connecting both insect cell bioreactor and mammalian cell bioreactor. Large amount of baculovirus will be produced in bcaulovirus/insect cell system. The fed-batch cultivation strategy will be investigate to achieve high virus density for high MOI needed in baculovirus/mammalian cell bioreactor. We will also investigate the productivity and efficiency of heterologous protein production as Granulocyte Macrophage Colony Stimulating Factor (GM-CSF) by the combination of GM-CSF foreign gene and this baculovirus/mammalian cell bioreactor.

Project ID:PB10207-1809
External Project ID:NSC102-2221-E182-050