Influence des conditions de culture sur la quantité de l'INF-[gamma] recombinant produit par des cellules CHO au cours de procédés discontinus

Abstract : In this study, we characterized the effect of culture conditions on the quantity and the quality of a recombinant protein, IFN-g, produced by CHO cells. In particular, we studied the effect of 3 components (iron citrate, pluronic F-68 and ethanolamine) that are present in the BDM medium, but completely lacking in RPMI serum medium (FCS-RPMI) on CHO cell growth, as well as the production and quality of recombinant IFN-g.The addition of Pluronic F-68 (0.1%) and iron citrate (500 µM) in RPMI without serum resulted in growth kinetic performances similar to those observed in FCS-RPMI. Furthermore, in these culture conditions, IFN- ? production was improved. Addition of iron citrate in FCS-RPMI improved cell growth, as well as IFN-g production. Whereas the glycosylation pattern of recombinant IFN-g produced by CHO cells was not constant when the culture was performed in FCS-RPMI, the glycosylation pattern of IFN-g remained constant when iron citrate was added in the medium. In addition, gelatinase and caseinase enzymatic activities in CHO batch cultures were detected, due most likely to enzymes of the metalloproteases and serine protease families. Despite the type of medium used (RPMI, BDM with or without serum), addition of iron citrate minimized IFN-g proteolysis. Finally, the relationship between the macroglycosylation pattern of IFN-g and its in-vitro biological (immunomodulatory) activity was demonstrated.
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Marie-Françoise Clincke. Influence des conditions de culture sur la quantité de l'INF-[gamma] recombinant produit par des cellules CHO au cours de procédés discontinus. Alimentation et Nutrition. Institut National Polytechnique de Lorraine, 2010. Français. ⟨NNT : 2010INPL034N⟩. ⟨tel-01749449⟩

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