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Long-Term Culturing of FreeStyle 293-F Cells Affects Immunoglobulin G Glycome Composition

Glycosylation of IgG regulates the effector function of this antibody in the immune response. Glycosylated IgG is a potent therapeutic used for both research and clinical purposes. While there is ample research on how different cell culture conditions affect IgG glycosylation, the data are missing o...

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Autores principales: Lukšić, Fran, Mijakovac, Anika, Josipović, Goran, Vičić Bočkor, Vedrana, Krištić, Jasminka, Cindrić, Ana, Vinicki, Martina, Rokić, Filip, Vugrek, Oliver, Lauc, Gordan, Zoldoš, Vlatka
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10452533/
https://www.ncbi.nlm.nih.gov/pubmed/37627310
http://dx.doi.org/10.3390/biom13081245
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author Lukšić, Fran
Mijakovac, Anika
Josipović, Goran
Vičić Bočkor, Vedrana
Krištić, Jasminka
Cindrić, Ana
Vinicki, Martina
Rokić, Filip
Vugrek, Oliver
Lauc, Gordan
Zoldoš, Vlatka
author_facet Lukšić, Fran
Mijakovac, Anika
Josipović, Goran
Vičić Bočkor, Vedrana
Krištić, Jasminka
Cindrić, Ana
Vinicki, Martina
Rokić, Filip
Vugrek, Oliver
Lauc, Gordan
Zoldoš, Vlatka
author_sort Lukšić, Fran
collection PubMed
description Glycosylation of IgG regulates the effector function of this antibody in the immune response. Glycosylated IgG is a potent therapeutic used for both research and clinical purposes. While there is ample research on how different cell culture conditions affect IgG glycosylation, the data are missing on the stability of IgG glycome during long cell passaging, i.e., cell “aging”. To test this, we performed three independent time course experiments in FreeStyle 293-F cells, which secrete IgG with a human-like glycosylation pattern and are frequently used to generate defined IgG glycoforms. During long-term cell culturing, IgG glycome stayed fairly stable except for galactosylation, which appeared extremely variable. Cell transcriptome analysis revealed no correlation in galactosyltransferase B4GALT1 expression with galactosylation change, but with expression of EEF1A1 and SLC38A10, genes previously associated with IgG galactosylation through GWAS. The FreeStyle 293-F cell-based system for IgG production is a good model for studies of mechanisms underlying IgG glycosylation, but results from the present study point to the utmost importance of the need to control IgG galactosylation in both in vitro and in vivo systems. This is especially important for improving the production of precisely glycosylated IgG for therapeutic purposes, since IgG galactosylation affects the inflammatory potential of IgG.
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spelling pubmed-104525332023-08-26 Long-Term Culturing of FreeStyle 293-F Cells Affects Immunoglobulin G Glycome Composition Lukšić, Fran Mijakovac, Anika Josipović, Goran Vičić Bočkor, Vedrana Krištić, Jasminka Cindrić, Ana Vinicki, Martina Rokić, Filip Vugrek, Oliver Lauc, Gordan Zoldoš, Vlatka Biomolecules Article Glycosylation of IgG regulates the effector function of this antibody in the immune response. Glycosylated IgG is a potent therapeutic used for both research and clinical purposes. While there is ample research on how different cell culture conditions affect IgG glycosylation, the data are missing on the stability of IgG glycome during long cell passaging, i.e., cell “aging”. To test this, we performed three independent time course experiments in FreeStyle 293-F cells, which secrete IgG with a human-like glycosylation pattern and are frequently used to generate defined IgG glycoforms. During long-term cell culturing, IgG glycome stayed fairly stable except for galactosylation, which appeared extremely variable. Cell transcriptome analysis revealed no correlation in galactosyltransferase B4GALT1 expression with galactosylation change, but with expression of EEF1A1 and SLC38A10, genes previously associated with IgG galactosylation through GWAS. The FreeStyle 293-F cell-based system for IgG production is a good model for studies of mechanisms underlying IgG glycosylation, but results from the present study point to the utmost importance of the need to control IgG galactosylation in both in vitro and in vivo systems. This is especially important for improving the production of precisely glycosylated IgG for therapeutic purposes, since IgG galactosylation affects the inflammatory potential of IgG. MDPI 2023-08-14 /pmc/articles/PMC10452533/ /pubmed/37627310 http://dx.doi.org/10.3390/biom13081245 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Lukšić, Fran
Mijakovac, Anika
Josipović, Goran
Vičić Bočkor, Vedrana
Krištić, Jasminka
Cindrić, Ana
Vinicki, Martina
Rokić, Filip
Vugrek, Oliver
Lauc, Gordan
Zoldoš, Vlatka
Long-Term Culturing of FreeStyle 293-F Cells Affects Immunoglobulin G Glycome Composition
title Long-Term Culturing of FreeStyle 293-F Cells Affects Immunoglobulin G Glycome Composition
title_full Long-Term Culturing of FreeStyle 293-F Cells Affects Immunoglobulin G Glycome Composition
title_fullStr Long-Term Culturing of FreeStyle 293-F Cells Affects Immunoglobulin G Glycome Composition
title_full_unstemmed Long-Term Culturing of FreeStyle 293-F Cells Affects Immunoglobulin G Glycome Composition
title_short Long-Term Culturing of FreeStyle 293-F Cells Affects Immunoglobulin G Glycome Composition
title_sort long-term culturing of freestyle 293-f cells affects immunoglobulin g glycome composition
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10452533/
https://www.ncbi.nlm.nih.gov/pubmed/37627310
http://dx.doi.org/10.3390/biom13081245
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