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 | Hansen, Henning Gram | ko |
dc.contributor.author | Arnsdorf, Johnny | ko |
dc.contributor.author | Nallapareddy, Saranya | ko |
dc.contributor.author | Voldborg, Bjorn | 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 | 2019-02-21T01:24:38Z | - |
dc.date.available | 2019-02-21T01:24:38Z | - |
dc.date.created | 2019-02-18 | - |
dc.date.issued | 2019-03 | - |
dc.identifier.citation | METABOLIC ENGINEERING, v.52, pp.143 - 152 | - |
dc.identifier.issn | 1096-7176 | - |
dc.identifier.uri | http://hdl.handle.net/10203/250460 | - |
dc.description.abstract | Recombinant Chinese hamster ovary (CHO) cells are able to provide biopharmaceuticals that are essentially free of human viruses and have N-glycosylation profiles similar, but not identical, to humans. Due to differences in N-glycan moieties, two members of the serpin superfamily, alpha-1-antitrypsin (A1AT) and plasma protease C1 inhibitor (C1INH), are currently derived from human plasma for treating A1AT and C1INH deficiency. Deriving therapeutic proteins from human plasma is generally a cost-intensive process and also harbors a risk of transmitting infectious particles. Recombinantly produced A1AT and C1INH (rhA1AT, rhC1INH) decorated with humanized N-glycans are therefore of clinical and commercial interest. Here, we present engineered CHO cell lines producing rhA1AT or rhC1INH with fully humanized N-glycosylation profiles. This was achieved by combining CRISPR/Cas9-mediated disruption of 10 gene targets with overexpression of human ST6GAL1. We were able to show that the N-linked glyco-structures of rhA1AT and rhC1INH are homogeneous and similar to the structures obtained from plasma-derived A1AT and C1INH, marketed as Prolastin (R)-C and Cinryze (R), respectively. rhA1AT and rhC1INH produced in our glyco-engineered cell line showed no detectable differences to their plasma-purified counterparts on SDS-PAGE and had similar enzymatic in vitro activity. The work presented here shows the potential of expanding the glyco-engineering toolbox for CHO cells to produce a wider variety of glycoproteins with fully humanized N-glycan profiles. We envision replacing plasma-derived A1AT and C1INH with recombinant versions and thereby decreasing our dependence on human donor blood, a limited and possibly unsafe protein source for patients. | - |
dc.language | English | - |
dc.publisher | ACADEMIC PRESS INC ELSEVIER SCIENCE | - |
dc.title | Glyco-engineered CHO cell lines producing alpha-1-antitrypsin and C1 esterase inhibitor with fully humanized N-glycosylation profiles | - |
dc.type | Article | - |
dc.identifier.wosid | 000457633200014 | - |
dc.identifier.scopusid | 2-s2.0-85058065150 | - |
dc.type.rims | ART | - |
dc.citation.volume | 52 | - |
dc.citation.beginningpage | 143 | - |
dc.citation.endingpage | 152 | - |
dc.citation.publicationname | METABOLIC ENGINEERING | - |
dc.identifier.doi | 10.1016/j.ymben.2018.11.014 | - |
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 | Hansen, Henning Gram | - |
dc.contributor.nonIdAuthor | Arnsdorf, Johnny | - |
dc.contributor.nonIdAuthor | Nallapareddy, Saranya | - |
dc.contributor.nonIdAuthor | Voldborg, Bjorn | - |
dc.contributor.nonIdAuthor | Andersen, Mikael Rordam | - |
dc.contributor.nonIdAuthor | Kildegaard, Helene Faustrup | - |
dc.description.isOpenAccess | N | - |
dc.type.journalArticle | Article | - |
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