Can I leave my Expi293F culture expressing for more than 7 days?
The growth and expression characteristics of Expi293F cells are such that, by 7 days post-transfection, the culture medium should be close to being spent and maximal protein expression should have already been achieved. Continued incubation will result in a large decrease in cell culture viability.
Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.
How many days should I wait before harvesting my protein in the Expi293 Expression System?
The optimal expression time will be different for each protein, but most tested proteins fall within a 3-7 day window. As the system scales well, it is recommended you run a small-scale pilot experiment to determine when to harvest your protein of interest prior to scaling up.
Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.
If I let my Expi293F cells grow to a higher density than recommended, will they be okay?
While the Expi293 Expression Medium can support much higher cell densities, we do not recommend growing your Expi293F cultures beyond 5-6 x 10e6 cells/mL, as subsequent transfection and protein expression efficiencies may be reduced. At higher densities, there is also the increased possibility of reaching the point of reduced culture viability. If your seed culture does exceed 5-6 x 10e6 cells/mL, passage them once or twice as detailed in the manual, monitoring them for viability and growth rate. Perform a test transfection using the Protein Expression Control IgG or an expression construct of known yield to determine if cell expression performance has been impacted.
Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.
Can I use a different 293 cell line with the Expi293 Expression System?
Other 293 cell lines may be used with the Expi293 Expression System. However, before these cell lines may be used for transfection studies, they must be adapted to serum-free, suspension culture in Expi293 Expression Medium and evaluated for transfection and expression.
Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.
Which vector do you recommend using with the Expi293 Expression System?
We recommend using the pcDNA 3.4-TOPO TA vector (Cat. No. A14697). This vector contains the native, full-length CMV promoter and a WPRE (Woodchuck Posttranscriptional Regulatory Element) downstream of the cloning site, both of which contribute to high level gene expression (about 2-3 fold higher expression than with pcDNA3.3 TOPO vector, which in turn provides 2-5 fold higher expression than with a standard pcDNA vector). Of course, the expression level is also protein-dependent.
Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.