Compartment‐specific 13C metabolic flux analysis reveals boosted NADPH availability coinciding with increased cell‐specific productivity for IgG1 producing CHO cells after MTA treatment

Abstract Increasing cell‐specific productivities (CSPs) for the production of heterologous proteins in Chinese hamster ovary (CHO) cells is an omnipresent need in the biopharmaceutical industry. The novel additive 5′‐deoxy‐5′‐(methylthio)adenosine (MTA), a chemical degradation product of S‐(5′‐adeno...

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Autores principales:	Andy Wiranata Wijaya, Natascha Verhagen, Attila Teleki, Ralf Takors
Formato:	article
Lenguaje:	EN
Publicado:	Wiley-VCH 2021
Materias:	13C MFA cell‐specific productivity CHO MTA NADPH Biotechnology TP248.13-248.65
Acceso en línea:	https://doaj.org/article/d171d3039cfe4829ada3ee91d61b3840
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spelling	oai:doaj.org-article:d171d3039cfe4829ada3ee91d61b38402021-12-02T13:20:01ZCompartment‐specific 13C metabolic flux analysis reveals boosted NADPH availability coinciding with increased cell‐specific productivity for IgG1 producing CHO cells after MTA treatment1618-28631618-024010.1002/elsc.202100057https://doaj.org/article/d171d3039cfe4829ada3ee91d61b38402021-12-01T00:00:00Zhttps://doi.org/10.1002/elsc.202100057https://doaj.org/toc/1618-0240https://doaj.org/toc/1618-2863Abstract Increasing cell‐specific productivities (CSPs) for the production of heterologous proteins in Chinese hamster ovary (CHO) cells is an omnipresent need in the biopharmaceutical industry. The novel additive 5′‐deoxy‐5′‐(methylthio)adenosine (MTA), a chemical degradation product of S‐(5′‐adenosyl)‐ʟ‐methionine (SAM) and intermediate of polyamine biosynthesis, boosts the CSP of IgG1‐producing CHO cells by 50%. Compartment‐specific 13C flux analysis revealed a fundamental reprogramming of the central metabolism after MTA addition accompanied by cell‐cycle arrest and increased cell volumes. Carbon fluxes into the pentose‐phosphate pathway increased 22 fold in MTA‐treated cells compared to that in non‐MTA‐treated reference cells. Most likely, cytosolic ATP inhibition of phosphofructokinase mediated the carbon detour. Mitochondrial shuttle activity of the α‐ketoglurarate/malate antiporter (OGC) reversed, reducing cytosolic malate transport. In summary, NADPH supply in MTA‐treated cells improved three fold compared to that in non‐MTA‐treated cells, which can be regarded as a major factor for explaining the boosted CSPs.Andy Wiranata WijayaNatascha VerhagenAttila TelekiRalf TakorsWiley-VCHarticle13C MFAcell‐specific productivityCHOMTANADPHBiotechnologyTP248.13-248.65ENEngineering in Life Sciences, Vol 21, Iss 12, Pp 832-847 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	13C MFA cell‐specific productivity CHO MTA NADPH Biotechnology TP248.13-248.65
spellingShingle	13C MFA cell‐specific productivity CHO MTA NADPH Biotechnology TP248.13-248.65 Andy Wiranata Wijaya Natascha Verhagen Attila Teleki Ralf Takors Compartment‐specific 13C metabolic flux analysis reveals boosted NADPH availability coinciding with increased cell‐specific productivity for IgG1 producing CHO cells after MTA treatment
description	Abstract Increasing cell‐specific productivities (CSPs) for the production of heterologous proteins in Chinese hamster ovary (CHO) cells is an omnipresent need in the biopharmaceutical industry. The novel additive 5′‐deoxy‐5′‐(methylthio)adenosine (MTA), a chemical degradation product of S‐(5′‐adenosyl)‐ʟ‐methionine (SAM) and intermediate of polyamine biosynthesis, boosts the CSP of IgG1‐producing CHO cells by 50%. Compartment‐specific 13C flux analysis revealed a fundamental reprogramming of the central metabolism after MTA addition accompanied by cell‐cycle arrest and increased cell volumes. Carbon fluxes into the pentose‐phosphate pathway increased 22 fold in MTA‐treated cells compared to that in non‐MTA‐treated reference cells. Most likely, cytosolic ATP inhibition of phosphofructokinase mediated the carbon detour. Mitochondrial shuttle activity of the α‐ketoglurarate/malate antiporter (OGC) reversed, reducing cytosolic malate transport. In summary, NADPH supply in MTA‐treated cells improved three fold compared to that in non‐MTA‐treated cells, which can be regarded as a major factor for explaining the boosted CSPs.
format	article
author	Andy Wiranata Wijaya Natascha Verhagen Attila Teleki Ralf Takors
author_facet	Andy Wiranata Wijaya Natascha Verhagen Attila Teleki Ralf Takors
author_sort	Andy Wiranata Wijaya
title	Compartment‐specific 13C metabolic flux analysis reveals boosted NADPH availability coinciding with increased cell‐specific productivity for IgG1 producing CHO cells after MTA treatment
title_short	Compartment‐specific 13C metabolic flux analysis reveals boosted NADPH availability coinciding with increased cell‐specific productivity for IgG1 producing CHO cells after MTA treatment
title_full	Compartment‐specific 13C metabolic flux analysis reveals boosted NADPH availability coinciding with increased cell‐specific productivity for IgG1 producing CHO cells after MTA treatment
title_fullStr	Compartment‐specific 13C metabolic flux analysis reveals boosted NADPH availability coinciding with increased cell‐specific productivity for IgG1 producing CHO cells after MTA treatment
title_full_unstemmed	Compartment‐specific 13C metabolic flux analysis reveals boosted NADPH availability coinciding with increased cell‐specific productivity for IgG1 producing CHO cells after MTA treatment
title_sort	compartment‐specific 13c metabolic flux analysis reveals boosted nadph availability coinciding with increased cell‐specific productivity for igg1 producing cho cells after mta treatment
publisher	Wiley-VCH
publishDate	2021
url	https://doaj.org/article/d171d3039cfe4829ada3ee91d61b3840
work_keys_str_mv	AT andywiranatawijaya compartmentspecific13cmetabolicfluxanalysisrevealsboostednadphavailabilitycoincidingwithincreasedcellspecificproductivityforigg1producingchocellsaftermtatreatment AT nataschaverhagen compartmentspecific13cmetabolicfluxanalysisrevealsboostednadphavailabilitycoincidingwithincreasedcellspecificproductivityforigg1producingchocellsaftermtatreatment AT attilateleki compartmentspecific13cmetabolicfluxanalysisrevealsboostednadphavailabilitycoincidingwithincreasedcellspecificproductivityforigg1producingchocellsaftermtatreatment AT ralftakors compartmentspecific13cmetabolicfluxanalysisrevealsboostednadphavailabilitycoincidingwithincreasedcellspecificproductivityforigg1producingchocellsaftermtatreatment
_version_	1718393297261559808

Compartment‐specific 13C metabolic flux analysis reveals boosted NADPH availability coinciding with increased cell‐specific productivity for IgG1 producing CHO cells after MTA treatment

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