title: Antioxidant effect of thiazolidine molecules in cell culture media improves stability and performance.
abstract: The ability of cell culture media components to generate reactive species as well as their sensitivity to oxidative degradation, affects the overall stability of media and the behavior of cells cultured in vitro. This study investigates the influence of thiazolidine molecules, formed from the condensation between cysteine and alpha-ketoacids, on the stability of these complex mixtures and on the performance of cell culture processes aiming to produce therapeutically relevant monoclonal antibodies. Results presented in this study indicate that 2-methyl-1,3-thiazolidine-2,4-dicarboxylic acid and 2-(2-carboxyethyl)-1,3-thiazolidine-2,4-dicarboxylic acid, obtained by condensation of cysteine with pyruvate or alpha-ketoglutarate, respectively, are able to stabilize cell culture media formulations, in particular redox sensitive molecules like folic acid, thiamine, l-methionine (met) and l-tryptophan (trp). The use of thiazolidine containing feeds in Chinese hamster ovary fed-batch processes showed prolonged culture duration and increased productivity. This enhanced performance was correlated with lower reactive species generation, extracellularly and intracellularly. Moreover, an anti-oxidative response was triggered via the induction of superoxide dismutase and an increase in the total glutathione pool, the major intracellular antioxidant. In total, the results confirm that cells in vitro are not cultured in an oxidant-free environment, a concept that has to be considered when studying the influence of reactive species in human diseases. Furthermore, this study indicates that thiazolidines are an interesting class of antioxidant molecules, capable of increasing cell culture media stability and process performance. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:759-770, 2017.
authors.affiliation: Merck Life Sciences, Upstream R&D, Frankfurter Strasse 250, Darmstadt, 64293, Germany.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Kuschelewski
authors.affiliation: Merck Life Sciences, Upstream R&D, Frankfurter Strasse 250, Darmstadt, 64293, Germany.
authors.forename: Alisa
authors.initials: A
authors.lastname: Schnellbaecher
authors.affiliation: Merck Life Sciences, Upstream R&D, Frankfurter Strasse 250, Darmstadt, 64293, Germany.
authors.forename: Sascha
authors.initials: S
authors.lastname: Pering
authors.affiliation: Merck Life Sciences, Upstream R&D, Frankfurter Strasse 250, Darmstadt, 64293, Germany.
authors.forename: Maria
authors.initials: M
authors.lastname: Wehsling
authors.affiliation: Merck Life Sciences, Upstream R&D, Frankfurter Strasse 250, Darmstadt, 64293, Germany.
authors.forename: Aline
authors.identifier: http://orcid.org/0000-0003-0279-4863
authors.initials: A
authors.lastname: Zimmer
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D016466:CHO Cells; D003412:Cricetulus; D003470:Culture Media; D005492:Folic Acid / Q000378:metabolism; D008715:Methionine / Q000378:metabolism; D013482:Superoxide Dismutase / Q000378:metabolism; D013831:Thiamine / Q000378:metabolism; D053778:Thiazolidines / Q000378:metabolism*; D014364:Tryptophan / Q000378:metabolism
chemical_list: D000975:Antioxidants; D003470:Culture Media; D053778:Thiazolidines; D014364:Tryptophan; D005492:Folic Acid; D008715:Methionine; D013482:Superoxide Dismutase; D013831:Thiamine
keywords: antioxidant; cell culture media reactivity and stability; fed-batch; productivity; thiazolidine