FreeStyle MAX System

Human IgG, Factor IX, human growth hormone (HGH), and erythropoietin (EPO) were produced in FreeStyle CHO-S cells. Cells were cultured to a density of 1 x 10⁶ cells/ml in 30 ml of FreeStyle CHO Expression Medium in a 125 ml shaker flask. The transfection complexes were formed using 1.25 μg of plasmid DNA and 1.25 μl of FreeStyle MAX Reagent per ml of culture volume to be transfected, in OptiPRO SFM. At 24 hours post-transfection, transfection efficiency was 60% as measured by GFP expression. Protein production was measured by ELISA, in mg/L.

The FreeStyle 293 System for easily scalable, rapid protein production generates functional, posttranslationally modified, proteins in human HEK293 cells. The system offers exceptional productivity and speed in the transient expression of recombinant antibody proteins (Table 2). Transfection is performed using the high-efficiency 293fectin transfection reagent. The transfection protocol can be readily scaled with minimal or no modifications for suspension culture volumes from less than 30 milliliters to greater than 10 liters while maintaining equivalent volumetric protein yields (Figure 3).

The FreeStyle 293 System kit includes 293fectin transfection reagent, Gibco FreeStyle 293 Expression Media, FreeStyle 293 cells, and Opti-MEM I medium. You can also purchase the system components separately.

Human IgG, Factor IX, human growth hormone (HGH), and erythropoietin (EPO) were produced in FreeStyle 293-F cells. Cells were cultured to a density of 1 x 10⁶ cells/ml in 30 ml of FreeStyle 293 Expression Medium in a 125 ml shaker flask. The transfection complexes were formed using 1.0 μg of plasmid DNA and 2.0 μl of 293fectin reagent per ml of culture volume to be transfected, in OptiPRO SFM or Opti-MEM I medium. At 24 hours post-transfection, transfection efficiency was >75% as measured by GFP expression. Protein production was measured by ELISA, in mg/L.

Large-scale, transient transfection of mammalian cells in suspension culture is a reliable and rapid method to generate milligram quantities of mammalian recombinant proteins for functional, therapeutic, and structural studies. With the FreeStyle MAX and FreeStyle 293 System,  you only need a sterile work bench, a CO₂ incubator, and a platform shaker with shake flasks or a spinner culture. Stable cell line development requires labor-intensive screening and adaptation procedures to identify highly expressing clones.

FreeStyle Custom Services are also available to produce your proteins under optimized conditions in suspension CHO or 293 systems based on the scale you request. We offer time-course studies, and protein production in 1–50 L cultures. We can provide you with the cell pellet, or even better, the purified protein, as the final deliverable. Contact your Invitrogen Sales Representative to discuss your needs and to initiate a project.