Transient Mammalian Cell Expression
Recombinant Protein Expression
Whether you are focused on lead discovery, production of targets for high throughput screenings, or characterization of interesting gene products or antibodies, our proprietary One-stop transient expression system will help optimize the expression and purification of your products.
Systems: Serum-free Suspension HEK293 and CHO cells.
Scales:50ml-1 liter for Pilot trial; 1-100 liter for wave bioreactor production
Fully customized services.
Fig 1. Semi-Quantitative SDS-PAGE of GFP-St6Gal1. GFP-human ST6Gal1 expressed in HEK293F suspension cells with freestyle serum-free medium. Plasmid DNA was transfected into HEK293F cells with 293fectin reagent. Supernatant were directly loaded on 12% SDS-PAGE gel (20ul). lane 1, 2, 3 and 4, 293fectin trasfected supernatant, dilution 1:2, 1:3, 1:4 and 1:6. lane 5, PEI transfected supernatant, no dilution. lane 6, 7 and 8, Purified TEV protease as standard, 100ug/ml, 50ug/ml and 25ug/ml.
- sub-clone your gene-of-interest into our proprietary mammalian expression vector. Codon optimization if necessary.
- Midi- or Maxi-prep of plasmid DNA with endotoxin-free kit.
- Transfect 50ml-1 liter of either HEK293 or CHO serum-free suspension cells. Scale is dependent on customer's choice.
- Analyze expression products by western blot, ELISA or purification and quantify protein production.
- Giga plasmid DNA Prep with endotoxin-free kit.
- Transfection of HEK293 or CHO at scales from 1 liter to 100 liter as customer needs.
- Culture 7-14 days depending on the cell viability.
- Harvest cells, clarification of supernatant, ultrafiltration to concentrate supernatant and to replace with purification compatible buffer.
- Load onto the affinity column. Further polish the eluted protein if necessary.
- Quality control, SDS-PAGE with coomassie blue staining or more if requested.