Expression of a Secreted Protein in Mammalian Cells Using Baculovirus Particles

  • Barbara Ann Jardin
  • Cynthia B. EliasEmail author
  • Satya Prakash
Part of the Methods in Molecular Biology book series (MIMB, volume 801)


There are many methods presently available to produce recombinant proteins in mammalian systems. The BacMam system is a simple straightforward method which overlaps two well-established technologies, namely the BEVS insect cell system and the transduction of mammalian cells in vitro. This chapter describes a method for the study of gene expression in mammalian cells in a series of simple steps. Protocols outlined include the design and construction of the recombinant baculovirus, cell culture techniques required to maintain both insect and mammalian cells, generation of baculovirus stocks, and methods to obtain maximal and reproducible gene expression in mammalian cells. Currently available statistical techniques using factorial design of experiment to optimize conditions for recombinant protein in vitro are outlined. Then details with respect to process scale-up in disposable bioreactors are included.

Key words

CMV promoter BacMam Baculovirus Full factorial design Design of experiment Hollow fiber Tangential flow filtration 


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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Barbara Ann Jardin
    • 1
    • 2
  • Cynthia B. Elias
    • 3
    Email author
  • Satya Prakash
    • 1
    • 2
  1. 1.Biomedical Technology and Cell Therapy Research Laboratory, Department of Biomedical Engineering, Artificial Cells and Organs Research CenterMcGill UniversityMontrealCanada
  2. 2.Biomedical Technology and Cell Therapy Research Laboratory, Department of Physiology, Artificial Cells and Organs Research CenterMcGill UniversityMontrealCanada
  3. 3.Bulk Manufacturing/Cells and Viral Media, Sanofi-PasteurTorontoCanada

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