Intérêts des hydrolysats de levure dans les procédés de culture de cellules CHO productrices d'anticorps : analyse cinétique, fractionnements et caractérisation des composés actifs

Abstract : This work studies the interest of the addition of yeast hydrolysates in culture medium of CHO cell producing antibody, to determine the operating conditions and their role, but also to improve the characterization of active compounds. In this way, an integrated approach including kinetic studies, fractionation strategies and biochemical analysis of hydrolysates and of their fractions was developed. First, we showed that yeast hydrolysates exhibited various properties depending on their composition and the operating conditions. In addition, synergistic effects were observed with different hydrolysate mixtures. Besides, kinetic studies underlined that the positive influence of hydrolysates on cell growth is correlated with energetic metabolism improvement. Then, the biochemical nature and the role of active compounds were studied by the implementation of a nanofiltration process and the reconstitution of mixtures of molecules contained in a yeast extract (YE). The results highlighted the interest of di- and tri-peptides to supply energetic metabolism, and of non-nutritive molecules, exhibiting a molecular weight greater than 500 Da, to stimulate the specific cell growth rate. Finally, the retentate fraction of nanofiltrated YE was fractionated by various chromatographic processes to characterize the physico-chemical properties of active compounds. The effect of fractions on cell culture emphasized the positive effect of positively charged molecules, especially hydrophilic and cationic peptides, to stimulate the cell growth. Thus, our work provides important insights in yeast hydrolysate mechanisms on CHO cells and suggests procedures to simplify such a complex additive of media dedicated to mammalian cell culture
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Mathilde Mosser. Intérêts des hydrolysats de levure dans les procédés de culture de cellules CHO productrices d'anticorps : analyse cinétique, fractionnements et caractérisation des composés actifs. Alimentation et Nutrition. Université de Lorraine, 2012. Français. ⟨NNT : 2012LORR0170⟩. ⟨tel-01749367⟩

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