title: Low-protein medium affects the 293SF central metabolism during growth and infection with adenovirus.
abstract: In this study the metabolism of 293SF cells grown in serum-free and low-protein medium was analyzed. This cell line is known for its ability to replicate recombinant adenovirus, mainly used in gene therapy applications. A complete model composed of the main glycolytic, glutaminolytic, and amino acids pathways, as well as the internalization fluxes of certain compounds into the mitochondria, is used for metabolic flux calculations. The pentose-phosphate cycle is also added to the biochemical reactions set and was independently measured with labeled 14C-glucose. Different feeding strategies in two different media were analyzed with the model, and the theoretical ATP production was also calculated. The two media were similar in their glucose and amino acid composition, but one contained BSA at 1g/L whereas the other had a very low protein content. Use of low-protein medium resulted in up to fourfold higher adenoviral vector production. In this medium, glucose utilization was more efficient, as it entered the TCA cycle more efficiently. Also, lower glutamine and amino acids consumption were observed as well as lower lactate and ammonia production. This increased TCA activity led to a twofold higher ATP production in the low-protein medium.
authors.affiliation: Institut de recherche en biotechnologie, CNRC, 6100 avenue Royalmount, Montréal, Québec H4P 2R2, Canada.
authors.forename: I
authors.initials: I
authors.lastname: Nadeau
authors.forename: P A
authors.initials: PA
authors.lastname: Gilbert
authors.forename: D
authors.initials: D
authors.lastname: Jacob
authors.forename: M
authors.initials: M
authors.lastname: Perrier
authors.forename: A
authors.initials: A
authors.lastname: Kamen
mesh_terms: D000255:Adenosine Triphosphate / Q000096:biosynthesis; D000260:Adenoviruses, Human; D019149:Bioreactors; D002250:Carbon Radioisotopes / Q000378:metabolism; D002451:Cell Compartmentation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line / Q000166:cytology / Q000378:metabolism* / Q000821:virology*; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D006801:Humans; D014176:Protein Biosynthesis; D011506:Proteins / Q000378:metabolism*; D012680:Sensitivity and Specificity; D014764:Viral Proteins / Q000096:biosynthesis / Q000378:metabolism
chemical_list: D002250:Carbon Radioisotopes; D016895:Culture Media, Serum-Free; D011506:Proteins; D014764:Viral Proteins; D000255:Adenosine Triphosphate