Transient Gene Expression in Suspension HEK293-EBNA1 Cells

  • Methods Mol Biol. 2018:1850:1-16. doi: 10.1007/978-1-4939-8730-6_1.
Denis L'Abbé  1 ,  Louis Bisson  1 ,  Christian Gervais  1 ,  Eric Grazzini  1 ,  Yves Durocher  2
Affiliations
  • 1. Human Health Therapeutics Research Center, National Research Council Canada, 6100 Royalmount Avenue, Montreal, H4P 2R2, Quebec, Canada.
  • 2. Human Health Therapeutics Research Center, National Research Council Canada, 6100 Royalmount Avenue, Montreal, H4P 2R2, Quebec, Canada. [email protected].
PMID: 30242676 DOI: 10.1007/978-1-4939-8730-6_1
Abstract

Transient gene expression in human embryo kidney 293 (HEK293) cells is an established approach for the rapid production of large amounts of Recombinant proteins (r-proteins). Milligram to gram quantities of r-proteins can be typically obtained within less than 10 days following transfection. In this chapter, we describe a simple and robust transfection process of suspension-growing human embryo kidney 293 cells using two commercially available serum-free media and polyethylenimine as the transfection reagent. This chapter provides examples for the production and purification of a his-tagged Recombinant protein and two Monoclonal Antibodies.

Keywords
293-6E; Antibody; Deacylated PEI; IMAC; Polyethylenimine; Protein-A; Recombinant protein; Serum-free medium; Suspension culture; Transfection.