Effects of cysteine, asparagine, or glutamine limitations in Chinese hamster ovary cell batch and fed-batch cultures.
Chinese hamster ovary cells
amino acid
fed-batch
monoclonal antibody
Journal
Biotechnology progress
ISSN: 1520-6033
Titre abrégé: Biotechnol Prog
Pays: United States
ID NLM: 8506292
Informations de publication
Date de publication:
03 2020
03 2020
Historique:
received:
09
08
2019
revised:
20
11
2019
accepted:
02
12
2019
pubmed:
12
12
2019
medline:
8
6
2021
entrez:
12
12
2019
Statut:
ppublish
Résumé
Amino acid availability is a key factor that can be controlled to optimize the productivity of fed-batch cultures. To study amino acid limitation effects, a serum-free chemically defined basal medium was formulated to exclude the amino acids that became depleted in batch culture. The effect of limiting glutamine, asparagine, and cysteine on the cell growth, metabolism, antibody productivity, and product glycosylation was investigated in three Chinese hamster ovary (CHO) cell lines (CHO-DXB11, CHO-K1SV, and CHO-S). Cysteine limitation was detrimental to both cell proliferation and productivity for all three CHO cell lines. Glutamine limitation reduced growth but not cell specific productivity, whereas asparagine limitation had no significant effect on either growth or cell specific productivity. Neither glutamine nor asparagine limitation significantly affected antibody glycosylation. Replenishing the CHO-DXB11 culture with cysteine after 1 day of cysteine limitation allowed the cells to partially recover their growth and productivity. This recovery was not observed after 2 days of cysteine limitation. Based on these findings, a fed-batch protocol was developed using single or mixed amino acid supplementation. Although cell density and antibody concentration were lower compared to a commercial feed, the feeds based on cysteine supplementation yielded comparable cell specific productivity. Overall, this study showed that different amino acid limitations have varied effects on the performance of CHO cell cultures and that maintaining cysteine availability is a critical process parameter for the three cell lines investigated.
Substances chimiques
Immunoglobulin G
0
Glutamine
0RH81L854J
Asparagine
7006-34-0
Cysteine
K848JZ4886
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e2946Subventions
Organisme : NSERC
Pays : International
Organisme : NSERC MabNet
Pays : International
Informations de copyright
© 2019 American Institute of Chemical Engineers.
Références
Altamirano C, Paredes C, Illanes A, Cairó JJ, Gòdia F. Strategies for fed-batch cultivation of T-PA producing CHO cells: substitution of glucose and glutamine and rational design of culture medium. J Biotechnol. 2004;110(2):171-179.
Dowd JE, Kwok KE, Piret JM. Increased T-PA yields using ultrafiltration of an inhibitory product from CHO fed-batch culture. Biotechnol Prog. 2000;16(5):786-794.
Kim DY, Chaudhry MA, Kennard ML, et al. Fed-batch CHO cell T-PA production and feed glutamine replacement to reduce ammonia production. Biotechnol Prog. 2013;29(1):165-175.
Jardon MA, Sattha B, Braasch K, et al. Inhibition of glutamine-dependent autophagy increases t-PA production in CHO cell fed-batch processes. Biotechnol Bioeng. 2012;109(5):1228-1238.
Fomina-Yadlin D, Gosink JJ, McCoy R, et al. Cellular responses to individual amino-acid depletion in antibody expressing and parental CHO cell lines. Biotechnol Bioeng. 2014;111(5):965-979.
Salazar A, Keusgen M, von Hagen J. Amino acids in the cultivation of mammalian cells. Amino Acids. 2016;48:1161-1171.
Eagle H. Nutrition needs of mammalian cells in tissue culture. Science. 1955b;122(3168):501-504.
Häussinger D, Sies H. Glutamine Metabolism in Mammalian Tissues. Berlin, Heidelberg: Springer Berlin Heidelberg; 1984.
Fan Y, Jimenez Del Val I, Müller C, et al. A multi-pronged investigation into the effect of glucose starvation and culture duration on fed-batch CHO cell culture. Biotechnol Bioeng. 2015;112(10):2172-2184.
Berg JM, Tymoczko JL, Stryer L. Glucose can be synthesized from noncarbohydrate precursors. Biochemistry. 5th ed. New York: W H Freeman; 2002.
Milman HA, Cooney DA. The distribution of L-asparagine synthetase in the principal organs of several mammalian and avian species. Biochem J. 1974;142(1):27-35.
Arfin SM, Simpson DR, Chiang CS, Andrulis IL, Hatfield GW. A role for Asparaginyl-tRNA in the regulation of asparagine synthetase in a mammalian cell line. Proc Natl Acad Sci U S A. 1977;74(6):2367-2369.
Stipanuk MH, Dominy JE, Lee JI, Coloso RM. Mammalian cysteine metabolism: new insights into regulation of cysteine metabolism. J Nutr. 2006;136(6):1652-1659.
Handlogten MW, Lee-O'Brien A, Roy G, et al. Intracellular response to process optimization and impact on productivity and product aggregates for a high titer CHO cell process. Biotechnol Bioeng. 2018;115:126-138.
Bell A, Wang ZJ, Arbabi-Ghahroudi M, et al. Differential tumor-targeting abilities of three single-domain antibody formats. Cancer Lett. 2010;289(1):81-90.
Zhang J, Liu X, Bell A, et al. Transient expression and purification of chimeric heavy chain antibodies. Protein Expr Purif. 2009;65(1):77-82.
Kennard ML, Goosney DL, Monteith D, et al. The generation of stable, high MAb expressing CHO cell lines based on the artificial chromosome expression (ACE) technology. Biotechnol Bioeng. 2009;104(3):540-553.
Nasseri SS, Ghaffari G, Jardon MA, et al. Increased CHO cell fed-batch monoclonal antibody production using the autophagy inhibitor 3-MA or gradually increasing osmolality. Biochem Eng J. 2014;91:37-45.
Gervais A, Hammel YA, Pelloux S, et al. Glycosylation of human recombinant Gonadotrophins: characterization and batch-to-batch consistency. Glycobiology. 2003;13(3):179-189.
Liu B, Spearman M, Doering J, Lattová E, Perreault H, Butler M. The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. J Biotechnol. 2014;170:17-27.
Matés JM, Pérez-Gómez C, Núñez de Castro I, Asenjo M, Márquez J. Glutamine and its relationship with intracellular redox status, oxidative stress and cell proliferation/death. Int J Biochem Cell Biol. 2002;34(5):439-458.
Konorov SO, Jardon MA, Piret JM, Blades MW, Turner RF. Raman microspectroscopy of live cells under autophagy-inducing conditions. Analyst. 2012;137(20):4662-4668.
SM1 S, Phillips D, JP MC Jr, et al. The mammalian target of rapamycin (mTOR) pathway regulates mitochondrial oxygen consumption and oxidative capacity. J Biol Chem. 2006;281:27643-27652.
Lee JS, Lee GM. Rapamycin treatment inhibits CHO cell death in a serum-free suspension culture by autophagy induction. Biotechnol Bioeng. 2012;109(12):3093-3102.
Jiang J, Zuo Y, Gu Z. Rapamycin protects the mitochondria against oxidative stress and apoptosis in a rat model of Parkinson's disease. Int J Mol Med. 2013;31(4):825-832.
Kerksick C, Willoughby D. The antioxidant role of glutathione and N-acetyl-cysteine supplements and exercise-induced oxidative stress. J Int Soc Sports Nutr. 2005;2(2):38-44.
Pan X, Dalm C, Wijffels RH, Martens DE. Metabolic characterization of a CHO cell size increase phase in fed-batch cultures. Appl Microbiol Biotechnol. 2017;101:8101-8113.
Kshirsagar R, McElearney K, Gilbert A, Sinacore M, Ryll T. Controlling trisulfide modification in recombinant monoclonal antibody produced in fed-batch cell culture. Biotechnol Bioeng. 2012;109(10):2523-2532.