Optimization of a high-cell-density polyethylenimine transfection method for rapid protein production in CHO-EBNA1 cells

Publication date: 10 September 2018Source: Journal of Biotechnology, Volume 281Author(s): Matthew Stuible, Alina Burlacu, Sylvie Perret, Denis Brochu, Béatrice Paul-Roc, Jason Baardsnes, Martin Loignon, Eric Grazzini, Yves DurocherAbstractFor pre-clinical evaluation of biotherapeutic candidates, protein production by transient gene expression (TGE) in Chinese Hamster Ovary (CHO) cells offers important advantages, including the capability of rapidly and cost-effectively generating recombinant proteins that are highly similar to those produced in stable CHO clones. We have established a novel CHO clone (CHO-3E7) expressing a form of the Epstein-Barr virus nuclear antigen-1 (EBNA-1) with improved TGE productivity relative to parental CHO cells. Taking advantage of a new transfection-compatible media formulation that permits prolonged, high-density culture, we optimized transfection parameters (cell density, plasmid vector and polyethylenimine concentrations) and post-transfection culture conditions to establish a new, high-performing process for rapid protein production. The growth media is chemically defined, and a single hydrolysate feed is added post-transfection, followed by periodic glucose supplementation. This method gave significantly higher yields than our standard low-cell density, F17-based CHO-3E7 TGE method, averaging several hundred mg/l for a panel of recombinant proteins and antibodies. Purified antibodies produced using the two methods had distinct glycosylatio...
Source: Journal of Biotechnology - Category: Biotechnology Source Type: research