title: Expression of a secreted protein in mammalian cells using baculovirus particles.
abstract: 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.
authors: Biomedical Technology and Cell Therapy Research Laboratory, Department of Biomedical Engineering, Artificial Cells and Organs Research Center, McGill University, Montreal, QC, Canada.
authors: Barbara Ann
authors: BA
authors: Jardin
authors: Cynthia B
authors: CB
authors: Elias
authors: Satya
authors: S
authors: Prakash
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D019149:Bioreactors; D002460:Cell Line; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D007313:Insecta / Q000166:cytology; D008853:Microscopy; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D014161:Transduction, Genetic; D014771:Virion / Q000235:genetics*
chemical_list: D011994:Recombinant Proteins