The Effect of the Expression of the Antiapoptotic BHRF1 Gene on the Metabolic Behavior of a Hybridoma Cell Line

One of the most important limitations of mammalian cells-based bioprocesses, and particularly hybridoma cell lines, is the accelerated metabolism related to glucose and glutamine consumption. The high uptake rates of glucose and glutamine (i.e., the main sources of carbon, nitrogen and energy) lead...

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Autores: Martínez-Monge, Iván, Comas Sánchez, Pere, Catalán-Tatjer, David, Prat, Jordi, Casablancas, Antoni, Paredes, Carlos J., Lecina, Martí, Cairó, Jordi Joan
Tipo de recurso: artículo
Fecha de publicación:2021
País:España
Institución:Universitat Ramon Llull (URL)
Repositorio:DAU Arxiu Digital de la Universitat Ramon Llull
OAI Identifier:oai:dau.url.edu:20.500.14342/4408
Acceso en línea:http://hdl.handle.net/20.500.14342/4408
https://doi.org/10.3390/app11146258
Access Level:acceso abierto
Palabra clave:Hybridoma
Genome-scale metabolic model
Antiapoptotic gene
Mitochondrial transport
BHRF1
Hibridomes
Genòmica
575
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spelling The Effect of the Expression of the Antiapoptotic BHRF1 Gene on the Metabolic Behavior of a Hybridoma Cell LineMartínez-Monge, IvánComas Sánchez, PereCatalán-Tatjer, DavidPrat, JordiCasablancas, AntoniParedes, Carlos J.Lecina, MartíCairó, Jordi JoanHybridomaGenome-scale metabolic modelAntiapoptotic geneMitochondrial transportBHRF1HibridomesGenòmica575One of the most important limitations of mammalian cells-based bioprocesses, and particularly hybridoma cell lines, is the accelerated metabolism related to glucose and glutamine consumption. The high uptake rates of glucose and glutamine (i.e., the main sources of carbon, nitrogen and energy) lead to the production and accumulation of large amounts of lactate and ammonia in culture broth. Lactate and/or ammonia accumulation, together with the depletion of the main nutrients, are the major causes of apoptosis in hybridoma cell cultures. The KB26.5 hybridoma cell line, producing an IgG3, was engineered with BHRF1 (KB26.5-BHRF1), an Epstein–Barr virus-encoded early protein homologous to the antiapoptotic protein Bcl-2, with the aim of protecting the hybridoma cell line from apoptosis. Surprisingly, besides achieving effective protection from apoptosis, the expression of BHRF1 modified the metabolism of the hybridoma cell line. Cell physiology and metabolism analyses of the original KB26.5 and KB26.5-BHRF1 revealed an increase of cell growth rate, a reduction of glucose and glutamine consumption, as well as a decrease in lactate secretion in KB26.5-BHRF1 cells. A flux balance analysis allowed us to quantify the intracellular fluxes of both cell lines. The main metabolic differences were identified in glucose consumption and, consequently, the production of lactate. The lactate production flux was reduced by 60%, since the need for NADH regeneration in the cytoplasm decreased due to a more than 50% reduction in glucose uptake. In general terms, the BHRF1 engineered cell line showed a more efficient metabolism, with an increase in biomass volumetric productivity under identical culture conditions.info:eu-repo/semantics/publishedVersionMDPIUniversitat Ramon Llull. IQS202420242021info:eu-repo/semantics/article16 p.application/pdfhttp://hdl.handle.net/20.500.14342/4408https://doi.org/10.3390/app11146258reponame:DAU Arxiu Digital de la Universitat Ramon Llullinstname:Universitat Ramon Llull (URL)InglésApplied Sciencesinfo:eu-repo/grantAgreement/NNF/Grant number NNF10CC1016517/info:eu-repo/grantAgreement/NNF/Grant number NNF14OC0009473/© L'autor/aAttribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:dau.url.edu:20.500.14342/44082026-06-21T06:40:37Z
dc.title.none.fl_str_mv The Effect of the Expression of the Antiapoptotic BHRF1 Gene on the Metabolic Behavior of a Hybridoma Cell Line
title The Effect of the Expression of the Antiapoptotic BHRF1 Gene on the Metabolic Behavior of a Hybridoma Cell Line
spellingShingle The Effect of the Expression of the Antiapoptotic BHRF1 Gene on the Metabolic Behavior of a Hybridoma Cell Line
Martínez-Monge, Iván
Hybridoma
Genome-scale metabolic model
Antiapoptotic gene
Mitochondrial transport
BHRF1
Hibridomes
Genòmica
575
title_short The Effect of the Expression of the Antiapoptotic BHRF1 Gene on the Metabolic Behavior of a Hybridoma Cell Line
title_full The Effect of the Expression of the Antiapoptotic BHRF1 Gene on the Metabolic Behavior of a Hybridoma Cell Line
title_fullStr The Effect of the Expression of the Antiapoptotic BHRF1 Gene on the Metabolic Behavior of a Hybridoma Cell Line
title_full_unstemmed The Effect of the Expression of the Antiapoptotic BHRF1 Gene on the Metabolic Behavior of a Hybridoma Cell Line
title_sort The Effect of the Expression of the Antiapoptotic BHRF1 Gene on the Metabolic Behavior of a Hybridoma Cell Line
dc.creator.none.fl_str_mv Martínez-Monge, Iván
Comas Sánchez, Pere
Catalán-Tatjer, David
Prat, Jordi
Casablancas, Antoni
Paredes, Carlos J.
Lecina, Martí
Cairó, Jordi Joan
author Martínez-Monge, Iván
author_facet Martínez-Monge, Iván
Comas Sánchez, Pere
Catalán-Tatjer, David
Prat, Jordi
Casablancas, Antoni
Paredes, Carlos J.
Lecina, Martí
Cairó, Jordi Joan
author_role author
author2 Comas Sánchez, Pere
Catalán-Tatjer, David
Prat, Jordi
Casablancas, Antoni
Paredes, Carlos J.
Lecina, Martí
Cairó, Jordi Joan
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universitat Ramon Llull. IQS
dc.subject.none.fl_str_mv Hybridoma
Genome-scale metabolic model
Antiapoptotic gene
Mitochondrial transport
BHRF1
Hibridomes
Genòmica
575
topic Hybridoma
Genome-scale metabolic model
Antiapoptotic gene
Mitochondrial transport
BHRF1
Hibridomes
Genòmica
575
description One of the most important limitations of mammalian cells-based bioprocesses, and particularly hybridoma cell lines, is the accelerated metabolism related to glucose and glutamine consumption. The high uptake rates of glucose and glutamine (i.e., the main sources of carbon, nitrogen and energy) lead to the production and accumulation of large amounts of lactate and ammonia in culture broth. Lactate and/or ammonia accumulation, together with the depletion of the main nutrients, are the major causes of apoptosis in hybridoma cell cultures. The KB26.5 hybridoma cell line, producing an IgG3, was engineered with BHRF1 (KB26.5-BHRF1), an Epstein–Barr virus-encoded early protein homologous to the antiapoptotic protein Bcl-2, with the aim of protecting the hybridoma cell line from apoptosis. Surprisingly, besides achieving effective protection from apoptosis, the expression of BHRF1 modified the metabolism of the hybridoma cell line. Cell physiology and metabolism analyses of the original KB26.5 and KB26.5-BHRF1 revealed an increase of cell growth rate, a reduction of glucose and glutamine consumption, as well as a decrease in lactate secretion in KB26.5-BHRF1 cells. A flux balance analysis allowed us to quantify the intracellular fluxes of both cell lines. The main metabolic differences were identified in glucose consumption and, consequently, the production of lactate. The lactate production flux was reduced by 60%, since the need for NADH regeneration in the cytoplasm decreased due to a more than 50% reduction in glucose uptake. In general terms, the BHRF1 engineered cell line showed a more efficient metabolism, with an increase in biomass volumetric productivity under identical culture conditions.
publishDate 2021
dc.date.none.fl_str_mv 2021
2024
2024
dc.type.none.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.14342/4408
https://doi.org/10.3390/app11146258
url http://hdl.handle.net/20.500.14342/4408
https://doi.org/10.3390/app11146258
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Applied Sciences
info:eu-repo/grantAgreement/NNF/Grant number NNF10CC1016517/
info:eu-repo/grantAgreement/NNF/Grant number NNF14OC0009473/
dc.rights.none.fl_str_mv © L'autor/a
Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv © L'autor/a
Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 16 p.
application/pdf
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv reponame:DAU Arxiu Digital de la Universitat Ramon Llull
instname:Universitat Ramon Llull (URL)
instname_str Universitat Ramon Llull (URL)
reponame_str DAU Arxiu Digital de la Universitat Ramon Llull
collection DAU Arxiu Digital de la Universitat Ramon Llull
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,811543

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