100-liter transient transfection.

Area of Science:

• Biotechnology
• Bioprocessing
• Molecular Biology

Background:

• Traditional methods for recombinant protein production often involve time-consuming development of stable cell lines.
• There is a need for faster and more flexible methods to produce therapeutic proteins, such as monoclonal antibodies.

Purpose of the Study:

• To report the first successful large-scale transient gene expression (LS-TGE) in a 100 L bioreactor.
• To demonstrate the feasibility of producing gram quantities of monoclonal antibodies using LS-TGE.
• To highlight the speed and flexibility of LS-TGE compared to traditional methods.

Main Methods:

• Utilized suspension-adapted HEK 293 EBNA SF cells in a 150 L bioreactor.
• Employed a modified calcium phosphate co-precipitation method for transfection with over 75 mg of plasmid DNA.
• Co-transfected three plasmids encoding antibody heavy chain, light chain, and green fluorescent protein (GFP) for efficiency monitoring.

Main Results:

• Achieved successful transient transfection at the 100 L scale.
• Produced over half a gram of monoclonal antibody (IgG) in less than 10 days.
• Monitored transfection efficiency via GFP expression, detectable as early as 20 hours post-transfection.

Conclusions:

• LS-TGE is a viable technology for producing hundreds of milligrams to gram amounts of recombinant proteins at the 100 L scale.
• LS-TGE offers significant advantages in speed, economy, and flexibility, eliminating the need for stable cell line development.
• This technology enables rapid production campaigns for diverse recombinant proteins using a single host cell line and seed train.