caa a22 4500 60550640X CHVBK 20210128100627.0 cr unu---uuuuu 210128e20150601xx s 000 0 eng 10.1007/s00253-015-6514-4 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6514-4 Benchmarking of commercially available CHO cell culture media for antibody production [Elektronische Daten] [David Reinhart, Lukas Damjanovic, Christian Kaisermayer, Renate Kunert] In this study, eight commercially available, chemically defined Chinese hamster ovary (CHO) cell culture media from different vendors were evaluated in batch culture using an IgG-producing CHO DG44 cell line as a model. Medium adaptation revealed that the occurrence of even small aggregates might be a good indicator of cell growth performance in subsequent high cell density cultures. Batch experiments confirmed that the culture medium has a significant impact on bioprocess performance, but high amino acid concentrations alone were not sufficient to ensure superior cell growth and high antibody production. However, some key amino acids that were limiting in most media could be identified. Unbalanced glucose and amino acids led to high cell-specific lactate and ammonium production rates. In some media, persistently high glucose concentrations probably induced the suppression of respiration and oxidative phosphorylation, known as Crabtree effect, which resulted in high cell-specific glycolysis rates along with a continuous and high lactate production. In additional experiments, two of the eight basal media were supplemented with feeds from two different manufacturers in six combinations, in order to understand the combined impact of media and feeds on cell metabolism in a CHO fed-batch process. Cell growth, nutrient consumption and metabolite production rates, antibody production, and IgG quality were evaluated in detail. Concentrated feed supplements boosted cell concentrations almost threefold and antibody titers up to sevenfold. Depending on the fed-batch strategy, fourfold higher peak cell concentrations and eightfold increased IgG titers (up to 5.8g/L) were achieved. The glycolytic flux was remarkably similar among the fed-batches; however, substantially different specific lactate production rates were observed in the different media and feed combinations. Further analysis revealed that in addition to the feed additives, the basal medium can make a considerable contribution to the ammonium metabolism of the cells. The glycosylation of the recombinant antibody was influenced by the selection of basal medium and feeds. Differences of up to 50% in the monogalacto-fucosylated (G1F) and high mannose fraction of the IgG were observed. The Author(s), 2015 Chinese hamster ovary (CHO) nationallicence Cell culture medium nationallicence Antibody production nationallicence Batch nationallicence Fed-batch nationallicence Medium benchmarking nationallicence Reinhart David Vienna Institute of BioTechnology, Department of Biotechnology, University of Natural Resources and Life Sciences,Vienna, Muthgasse 11, 1190, Vienna, Austria aut Damjanovic Lukas Vienna Institute of BioTechnology, Department of Biotechnology, University of Natural Resources and Life Sciences,Vienna, Muthgasse 11, 1190, Vienna, Austria aut Kaisermayer Christian BioMarin International Limited, Shanbally, County Cork, Ringaskiddy, Ireland aut Kunert Renate Vienna Institute of BioTechnology, Department of Biotechnology, University of Natural Resources and Life Sciences,Vienna, Muthgasse 11, 1190, Vienna, Austria aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/11(2015-06-01), 4645-4657 0175-7598 99:11<4645 2015 99 253 https://doi.org/10.1007/s00253-015-6514-4 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6514-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Reinhart David Vienna Institute of BioTechnology, Department of Biotechnology, University of Natural Resources and Life Sciences,Vienna, Muthgasse 11, 1190, Vienna, Austria aut NATIONALLICENCE 700 1- Damjanovic Lukas Vienna Institute of BioTechnology, Department of Biotechnology, University of Natural Resources and Life Sciences,Vienna, Muthgasse 11, 1190, Vienna, Austria aut NATIONALLICENCE 700 1- Kaisermayer Christian BioMarin International Limited, Shanbally, County Cork, Ringaskiddy, Ireland aut NATIONALLICENCE 700 1- Kunert Renate Vienna Institute of BioTechnology, Department of Biotechnology, University of Natural Resources and Life Sciences,Vienna, Muthgasse 11, 1190, Vienna, Austria aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/11(2015-06-01), 4645-4657 0175-7598 99:11<4645 2015 99 253