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Coupling CRISPR interference with FACS enrichment [Elektronski vir] : new approach in glycoengineering of CHO cell lines for therapeutic glycoprotein productionGlinšek, Katja ...Difficulties in obtaining and maintaining the desired level of the critical quality attributes (CQAs) of therapeutic proteins as well as the pace of the development are major challenges of current ... biopharmaceutical development. Therapeutic proteins, both innovative and biosimilars, are mostly glycosylated. Glycans directly influence the stability, potency, plasma half-life, immunogenicity, and effector functions of the therapeutic. Hence, glycosylation is widely recognized as a process-dependent CQA of therapeutic glycoproteins. Due to the typically high heterogeneity of glycoforms attached to the proteins, control of glycosylation represents one of the most challenging aspects of biopharmaceutical development. Here, we explored a new glycoengineering approach in therapeutic glycoproteins development, which enabled us to achieve the targeted glycoprofile of the Fc-fusion protein in a fast manner. Coupling CRISPRi technology with lectin-FACS sorting enabled downregulation of the endogenous gene involved in fucosylation and further enrichment of CHO cells producing Fc-fusion proteins with reduced fucosylation levels. Enrichment of cells with targeted glycoprofile can lead to time-optimized clone screening and speed up cell line development. Moreover, the presented approach allows isolation of clones with varying levels of fucosylation, which makes it applicable to a broad range of glycoproteins differing in target fucosylation level.Source: Biotechnology journal [Elektronski vir]. - ISSN 1860-7314 (Vol. 17, iss. 7, art. e2100499, 2022, 13 str.)Type of material - e-articlePublish date - 2022Language - englishCOBISS.SI-ID - 112903939
Author
Glinšek, Katja |
Kramer, Lovro |
Krajnc, Aleksander |
Kranjc, Eva |
Pirher, Nina |
Marušič, Jaka |
Hellmann, Leon |
Podobnik, Barbara |
Štrukelj, Borut |
Ausländer, David |
Gaber, Rok
Topics
Beljkovine |
CHO cells |
cell line development |
fucosylation |
glycoengineering |
therapeutic glycoproteins |
celice CHO |
razvoj celične linije |
fukozilacija |
glikoinženiring |
terapevtski glikoproteini
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Database name | Field | Year |
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Links to authors' personal bibliographies | Links to information on researchers in the SICRIS system |
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Glinšek, Katja | 50713 |
Kramer, Lovro | 35467 |
Krajnc, Aleksander | 37474 |
Kranjc, Eva | 36423 |
Pirher, Nina | 26500 |
Marušič, Jaka | 91697 |
Hellmann, Leon | |
Podobnik, Barbara | 20681 |
Štrukelj, Borut | 07849 |
Ausländer, David | |
Gaber, Rok | 32254 |
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