Transient Gene Expression in Suspension HEK293-EBNA1 Cells

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.