Conclusions
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- The addition of Recombinant Insulin led to a substantial 1.7-fold increase in mAb (Trastuzumab) productivity in CHO-S cell culture.
- The insulin dose response varies depending on the media used. In certain media, such as media 1, the fold-increase rises with higher concentrations, while in other media, like media 1 and 2, lower insulin concentrations further enhance cell culture performance compared to higher concentrations. This highlights the significance of tailoring insulin levels to specific cell culture conditions for optimal performance.
Background
The large-scale production of therapeutic proteins, including mAbs, relies heavily on CHO cells. Choosing media that support CHO cell proliferation is critical for enhancing productivity and cost efficiency. The supplementation of catalog media with animal-free cell culture supplements can further improve cell performance.
This study shows the advantages of supplementing various chemically-defined media with Recombinant Insulin, to boost CHO-S cell proliferation and achieve high protein yield.
Study description
In this study, three commercially available chemically-defined media were supplemented with varying concentrations of Recombinant Insulin (0, 1, and 5 mg/L). Cell count and viability were determined using the Nucleocounter NC-3000 (Chemometec), and the concentration of Trastuzumab in the supernatant was assessed using the ELISA SHIKARI® Q-TRAS commercial kit (MatriksBiotek Laboratories), following the manufacturer’s instructions.
The addition of various concentrations of Recombinant Insulin to three different catalog cell culture media resulted in enhanced cell proliferation in all cases, with the lowest insulin concentration leading to the highest increase in cell proliferation (Fig. 1).
Furthermore, the presence of insulin improved Trastuzumab production in CHO-S cells across all three catalog media tested, with the highest increase (1.7-fold) observed in media 3, when 1 mg/L insulin was supplemented to the cell culture media (Fig. 2).
Figure 1: Insulin improves growth across three different catalog media. CHO-S cells were cultured in three different commercially available chemically-defined media with or without insulin. Viable cell density and viability were measured daily. Mean ± standard deviation from three biological replicates is represented.
Figure 2: Insulin boosts Trastuzumab production in all three catalog media tested. Fold-change in antibody yield in the presence of insulin compared to the media without insulin, across three commercially available media (Media 1-3). Values are presented as the mean ± SD (n=3).
