Recombinant Plasminogen Activator Inhibitor-1
S domain Serine Protease Domain
SFM Serum Free Media
SGE Stable Gene Expression
TGE Transient Gene Expression
t-PA Tissue Plasminogen Activator
VPA Valproic Acid
Optimization of recombinant protein production in TGE system
Eukaryotic expression systems are the most prevalently used hosts in recombinant protein production, due to their appropriate post translational modifications. Nowadays, almost 60-70% of recombinant proteins are successfully produced in CHO cells Chinese hamster ovary (CHO) cells are capable of providing the correct protein glycosylation, and adapted to suspension growth at high cell density in large bioreactors.
The optimization of bioprocess conditions towards improved growth profile and productivity yields is considered of great importance in biopharmaceutical manufacturing. Peptones as efficient sources of nutrients have been studied for their effect on media development. Nevertheless, their role on metabolic pathway is not well understood.
Plant-derived peptones can improve cell growth and enhance specific and volumetric productivity both in stable and transient expression systems [4-6] . However, the positive effect of peptones is cell type and also protein dependant and similar feeding strategies may cause different responses in different systems. Consequently, a peptone supplemented bioprocess needs to be optimized for a specific cell line or clone. In the present study the effect of different concentration of peptones on a recombinant CHO cell line grown in three serum-free suspension cultures was determined. Six peptones of different origin and available amino acid profile were investigated regarding their impact on cell growth, productivity and metabolic pathways changes. In optimized feeding strategies increases of 136% and 159% of volumetric productivity (for low-nutrient culture media) and 55% (for a high-nutrient culture media) were achieved. Furthermore, particular sources of peptones with specific amino acid profile, developed preferential results for each different culture media. Two peptones SoyA2SC and SoyE-110 were the only hydrolysates which showed production improvement in all three media. Peptones Casein Peptone Plus, Tryptone N1 and SoyA3SC showed different improving results based on their concentration for each individual basal media. The amino acid profile of peptones provided some guidance in identifying the most effective feeding strategies for recombinant CHO cells.

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