Pregled bibliografske jedinice broj: 833639
An omics approach to rational feed Enhancing growth in CHO cultures with NMR metabolomics and 2D-DIGE proteomics
An omics approach to rational feed Enhancing growth in CHO cultures with NMR metabolomics and 2D-DIGE proteomics // Journal of biotechnology, 234 (2016), 127-138 doi:10.1016/j.jbiotec.2016.07.027 (međunarodna recenzija, članak, znanstveni)
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Naslov
An omics approach to rational feed Enhancing growth in CHO cultures with NMR metabolomics and 2D-DIGE proteomics
Autori
Blondeel, Eric J.M. ; ... ; Igor Slivac ; ... ; Aucoin, Marc G.
Izvornik
Journal of biotechnology (0168-1656) 234
(2016);
127-138
Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni
Ključne riječi
CHO cells ; 2D-DIGE ; Proteomics ; NMR ; Metabolomics
Sažetak
Expression of recombinant proteins exerts stress on cell culture systems, affecting the expression ofendogenous proteins, and contributing to the depletion of nutrients and accumulation of waste metabo-lites. In this work, 2D-DIGE proteomics was employed to analyze differential expression of proteinsfollowing stable transfection of a Chinese Hamster Ovary (CHO) cell line to constitutively express a heavy-chain monoclonal antibody. Thirty-four proteins of significant differential expression were identifiedand cross-referenced with cellular functions and metabolic pathways to identify points of cell stress.Subsequently, 1D-1H NMR metabolomics experiments analyzed cultures to observe nutrient deple-tion and waste metabolite accumulations to further examine these cell stresses and pathways. Fromamong fifty metabolites tracked in time-course, eight were observed to be completely depleted from theproduction media, including: glucose, glutamine, proline, serine, cystine, asparagine, choline, and hypox-anthine, while twenty-three excreted metabolites were also observed to accumulate. The differentiallyexpressed proteins, as well as the nutrient depletion and accumulation of these metabolites correspondedwith upregulated pathways and cell systems related to anaplerotic TCA-replenishment, NADH/NADPHreplenishment, tetrahydrofolate cycle C1 cofactor conversions, limitations to lipid synthesis, and redoxmodulation. A nutrient cocktail was assembled to improve the growth medium and alleviate these cellstresses to achieve a ∼75% improvement to peak cell densities.
Izvorni jezik
Engleski
Znanstvena područja
Biotehnologija
Poveznice na cjeloviti tekst rada:
Pristup cjelovitom tekstu rada doi www.sciencedirect.com dx.doi.orgCitiraj ovu publikaciju:
Časopis indeksira:
- Current Contents Connect (CCC)
- Web of Science Core Collection (WoSCC)
- Science Citation Index Expanded (SCI-EXP)
- SCI-EXP, SSCI i/ili A&HCI
- Scopus
