Specific nutrient supplementation of defined serum-free medium for the improvement of CHO cells growth and t-PA production

Claudia Altamirano, Andrés Illanes, Rossana Canessa, Silvana Becerra

Resultado de la investigación: Contribución a una revistaArtículorevisión exhaustiva

18 Citas (Scopus)

Resumen

Recombinant CHO TF70R cells are able to grow and produce t-PA on serum-free medium BIOPRO1 (BioWhitaker Europe, Belgium). The purpose of the present study was to determine the effect of medium supplementation with vitamins, lipids, and specific amino acids on cell growth, t-PA production and biological functionality. Among vitamins, only biotin, folic acid, cobalamine and benzoic acid were required for improving growth and t-PA production. Lipid supplement allowed a significant increase cell concentration and t-PA specific activity and concentration, though its specific production rate decreased slightly. Medium supplementation with proline, serine and asparagine had also positive effects on cell growth. Besides, the addition of asparagine (even in the presence of glutamine) was essential for the production and biological quality of the t-PA. This systematic approach for media supplementation produced an increase in cell concentration around 100% and in t-PA production around 80%, with no detrimental effect on its biological activity. The effect of asparagine on t-PA production was unexpected and needs to be further studied. The above modifications of the production medium did not produce a significant effect on the metabolism of the main carbon and energy sources (glucose and glutamine) and the level of byproduct formation (lactate and ammonia).

Idioma originalInglés
Páginas (desde-hasta)61-67
Número de páginas7
PublicaciónElectronic Journal of Biotechnology
Volumen9
N.º1
DOI
EstadoPublicada - 15 ene 2006
Publicado de forma externa

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