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. 2022 Sep 14;14(9):1949. doi: 10.3390/pharmaceutics14091949

Figure 2.

Figure 2

Suspension culturing improves the predictability of protein production. (a) Adherent cells were transfected with 0.2 µg of pLuc per 96-well plate well. CPP/pDNA complexes were formed at CR3:1, and PEI complexes were formed at N/P20. The total reporter levels were determined 24 h post-transfection. N = 10 for treatment groups. (b) The suspension cells were transfected with 0.1 µg of pLuc per 96-well plate well. CPP/pDNA complexes were formed at CR2:1. Total reporter levels were determined 48 h post-transfection. N = 10 for treatment groups. (c) Adherent cells were transfected in serum-free media with 0.5 µg of green fluorescent protein encoding plasmid (pGFP) per 24-well plate well. 4 h post-transfection media was changed to serum-containing media. CPP/pDNA complexes were formed at CR3:1, PEI N/P20. Analysis was performed 24 h post-transfection. N ≥ 5. (d) Suspension cells were transfected in serum-free media with 0.75 µg of pGFP per 24-well plate well. CPP/pDNA complexes formed at CR2:1. For PEI N/P20 was used in case of the HEK293FT cells and N/P60 in case of CHO-K1 cells. 4 h post-transfection 500 µL of fresh media was added to each well. 48 h post-transfection the cells were analyzed by flow cytometry. N ≥ 6 for treatment groups.