Modulating antibody N-glycosylation through feed additives using a multi-tiered approach

Kranjc, Jaka (author), Kramer, Lovro (author), Mikelj, Miha (author), Anderluh, Marko (author), Pišlar, Anja (author), Brinc, Matjaž (author)

Glycosylation of recombinant proteins is a post-translational modification that affects multiple physicochemical and biological properties of proteins. As such, it is a critical quality attribute that must be carefully controlled during protein production in the pharmaceutical industry. Glycosylation can be modulated by various conditions, including the composition of production media and feeds. In this study, the N-glycosylation-modulating effects of numerous compounds, including metal enzyme cofactors, enzyme inhibitors, and metabolic intermediates, were evaluated. Chinese hamster ovary cells producing three different IgG antibodies were cultivated in a fed-batch mode. First, a one-factor-at-a-time experiment was performed in 24-well deep well plates to identify the strongest modulators and appropriate concentration ranges. Then, a full response surface experiment was designed to gauge the effects and interactions of the 14 most effective hit compounds in an Ambr® 15 bioreactor system. A wide range of glycoform content was achieved, with an up to eight-fold increase in individual glycoforms compared to controls. The resulting model can be used to determine modulator combinations that will yield desired glycoforms in the final product.

• Language: English
• Keywords: bioprocess, glycosylation, modulators, modelling, Chinese hamster ovary, antibody production
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: FFA - Faculty of Pharmacy
• Publication status: Published
• Publication version: Version of Record
• Year: 2024
• Number of pages: 13 str.
• Numbering: Vol. 12, [article no.] 1448925
• PID: 20.500.12556/RUL-175589
• UDC: 577.112
• ISSN on article: 2296-4185
• DOI: 10.3389/fbioe.2024.1448925
• COBISS.SI-ID: 246929411
• Publication date in RUL: 05.11.2025

Record is a part of a journal

• Title: Frontiers in bioengineering and biotechnology
• Shortened title: Front. bioeng. biotechnol.
• Publisher: Frontiers Editorial Office
• ISSN: 2296-4185
• COBISS.SI-ID: 523093017

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Secondary language

• Language: Slovenian
• Keywords: bioprocesi, glikozilacija, modulatorji, modeliranje, jajčnik kitajskega hrčka, proizvodnja protiteles, rekombinantni protein

Projects

• Funder: Other - Other funder or multiple funders
• Funding programme: Novartis Pharmaceutical Manufacturing LLC