The Use of Baculovirus-Mediated Gene Expression in Mammalian Cells for Recombinant Protein Production

Methods Mol Biol. 2024:2810:29-53. doi: 10.1007/978-1-0716-3878-1_3.

Abstract

Baculovirus-mediated gene expression in mammalian cells, BacMam, is a useful alternative to transient transfection for recombinant protein production in various types of mammalian cell lines. We decided to establish BacMam in our lab in order to streamline our workflows for gene expression in insect and mammalian cells, as it is straightforward to parallelize the baculovirus generation for both types of eukaryotic cells. This chapter provides a step-by-step description of the protocols we use for the generation of the recombinant BacMam viruses, the transduction of mammalian cell cultures, and optimization of the protein production conditions through small-scale expression and purification tests.

Keywords: BacMam; Baculovirus; Cell culture; Insect cells; Mammalian cells; Protein expression; Protein purification.

MeSH terms

  • Animals
  • Baculoviridae* / genetics
  • Cell Culture Techniques / methods
  • Cell Line
  • Gene Expression*
  • Genetic Vectors / genetics
  • Humans
  • Recombinant Proteins* / biosynthesis
  • Recombinant Proteins* / genetics
  • Recombinant Proteins* / metabolism
  • Sf9 Cells
  • Transduction, Genetic / methods
  • Transfection / methods

Substances

  • Recombinant Proteins