DC Field | Value | Language |
---|---|---|
dc.contributor.author | Amann, Thomas | ko |
dc.contributor.author | Hansen, Anders Holmgaard | ko |
dc.contributor.author | Kol, Stefan | ko |
dc.contributor.author | Lee, Gyun Min | ko |
dc.contributor.author | Andersen, Mikael Rordam | ko |
dc.contributor.author | Kildegaard, Helene Faustrup | ko |
dc.date.accessioned | 2018-10-19T00:53:40Z | - |
dc.date.available | 2018-10-19T00:53:40Z | - |
dc.date.created | 2018-10-15 | - |
dc.date.created | 2018-10-15 | - |
dc.date.issued | 2018-10 | - |
dc.identifier.citation | BIOTECHNOLOGY JOURNAL, v.13, no.10 | - |
dc.identifier.issn | 1860-6768 | - |
dc.identifier.uri | http://hdl.handle.net/10203/246208 | - |
dc.description.abstract | In production of recombinant proteins for biopharmaceuticals, N-glycosylation is often important for protein efficacy and patient safety. IgG with agalactosylated (G0)-N-glycans can improve the activation of the lectin-binding complement system and be advantageous in the therapy of lupus and virus diseases. In this study, the authors aimed to engineer CHO-S cells for the production of proteins with G0-N-glycans by targeting B4Gal-T isoform genes with CRISPR/Cas9. Indel mutations in genes encoding B4Gal-T1, -T2, and -T3 with and without a disrupted B4Gal-T4 sequence resulted in only approximate to 1% galactosylated N-glycans on total secreted proteins of 3-4 clones per genotype. The authors revealed that B4Gal-T4 is not active in N-glycan galactosylation in CHO-S cells. In the triple-KO clones, transiently expressed erythropoietin (EPO) and rituximab harbored only approximate to 6% and approximate to 3% galactosylated N-glycans, respectively. However, simultaneous disruption of B4Gal-T1 and -T3 may decrease cell growth. Altogether, the authors present the advantage of analyzing total secreted protein N-glycans after disrupting galactosyltransferases, followed by expressing recombinant proteins in selected clones with desired N-glycan profiles at a later stage. Furthermore, the authors provide a cell platform that prevalently glycosylates proteins with G0-N-glycans to further study the impact of agalactosylation on different in vitro and in vivo functions of recombinant proteins. | - |
dc.language | English | - |
dc.publisher | WILEY-V C H VERLAG GMBH | - |
dc.subject | CHO-CELLS | - |
dc.subject | GLYCOSYLATION MUTANTS | - |
dc.subject | IGG1 ANTIBODIES | - |
dc.subject | HIGH-MANNOSE | - |
dc.subject | LINES | - |
dc.subject | GENE | - |
dc.subject | OLIGOSACCHARIDES | - |
dc.subject | ERYTHROPOIETIN | - |
dc.subject | GENERATION | - |
dc.subject | EXPRESSION | - |
dc.title | CRISPR/Cas9-Multiplexed Editing of Chinese Hamster Ovary B4Gal-T1, 2, 3, and 4 Tailors N-Glycan Profiles of Therapeutics and Secreted Host Cell Proteins | - |
dc.type | Article | - |
dc.identifier.wosid | 000446009100012 | - |
dc.identifier.scopusid | 2-s2.0-85050493424 | - |
dc.type.rims | ART | - |
dc.citation.volume | 13 | - |
dc.citation.issue | 10 | - |
dc.citation.publicationname | BIOTECHNOLOGY JOURNAL | - |
dc.identifier.doi | 10.1002/biot.201800111 | - |
dc.contributor.localauthor | Lee, Gyun Min | - |
dc.contributor.nonIdAuthor | Amann, Thomas | - |
dc.contributor.nonIdAuthor | Hansen, Anders Holmgaard | - |
dc.contributor.nonIdAuthor | Kol, Stefan | - |
dc.contributor.nonIdAuthor | Andersen, Mikael Rordam | - |
dc.contributor.nonIdAuthor | Kildegaard, Helene Faustrup | - |
dc.description.isOpenAccess | N | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordAuthor | Chinese hamster ovary cells | - |
dc.subject.keywordAuthor | CRISPR | - |
dc.subject.keywordAuthor | Cas9 | - |
dc.subject.keywordAuthor | erythropoietin | - |
dc.subject.keywordAuthor | glycoengineering | - |
dc.subject.keywordAuthor | multiplexing | - |
dc.subject.keywordAuthor | N-glycosylation | - |
dc.subject.keywordAuthor | rituximab | - |
dc.subject.keywordPlus | CHO-CELLS | - |
dc.subject.keywordPlus | GLYCOSYLATION MUTANTS | - |
dc.subject.keywordPlus | IGG1 ANTIBODIES | - |
dc.subject.keywordPlus | HIGH-MANNOSE | - |
dc.subject.keywordPlus | LINES | - |
dc.subject.keywordPlus | GENE | - |
dc.subject.keywordPlus | OLIGOSACCHARIDES | - |
dc.subject.keywordPlus | ERYTHROPOIETIN | - |
dc.subject.keywordPlus | GENERATION | - |
dc.subject.keywordPlus | EXPRESSION | - |
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