Referenced in: none

Automatically associated channels: Kir2.3 , Kv7.2

Title: Transient gene expression: recombinant protein production with suspension-adapted HEK293-EBNA cells.

Authors: P Meissner, H Pick, A Kulangara, P Chatellard, K Friedrich, F M Wurm

Journal, date & volume: Biotechnol. Bioeng., 2001 Oct 20 , 75, 197-203

PubMed link: http://www.ncbi.nlm.nih.gov/pubmed/11536142

Abstract
Transient gene expression (TGE) in mammalian cells at the reactor scale is becoming increasingly important for the rapid production of recombinant proteins. We improved a process for transient calcium phosphate-based transfection of HEK293-EBNA cells in a 1-3 L bioreactor volume. Cells were adapted to suspension culture using a commercially available medium (BioWhittaker, Walkersville, MD). Process parameters were optimized using a plasmid reporter vector encoding the enhanced green fluorescent protein (EGFP/CLONTECH, Palo Alto, CA, USA). Using GFP as a marker-protein, we observed by microscopic examination transfection efficiencies between 70-100%. Three different recombinant proteins were synthesized within a timeframe of 7 days from time of transfection to harvest. The first, a human recombinant IgG(1)-type antibody, was secreted into the supernatant of the cell culture and achieved a final concentration of >20 mg/L. An E. coli-derived DNA-binding protein remained intracellular, as expected, but accumulated to such a concentration that the lysate of cells, taken up into the entire culture volume, gave a concentration of 18 mg/L. The third protein, a transmembrane receptor, was expressed at 3-6 x 10(6) molecules/cell.