Show simple item record

dc.contributor.authorThaysen-Andersen, Morten
dc.contributor.authorChertova, Elena N.
dc.contributor.authorBergamaschi, Cristina
dc.contributor.authorMoh, Edward S. X.
dc.contributor.authorChertov, Oleg Yu
dc.contributor.authorRoser, James D.
dc.contributor.authorSowder, Raymond C.
dc.contributor.authorBear, Jenifer S.
dc.contributor.authorLifson, Jeffrey D.
dc.contributor.authorPacker, Nicolle H.
dc.contributor.authorFelber, Barbara K.
dc.contributor.authorPavlakis, George N.
dc.date.accessioned2017-05-11T01:13:41Z
dc.date.available2017-05-11T01:13:41Z
dc.date.issued2016
dc.identifier.issn0282-0080
dc.identifier.doi10.1007/s10719-015-9627-1
dc.identifier.urihttp://hdl.handle.net/10072/336507
dc.description.abstractHuman interleukin 15 (IL-15) circulates in blood as a stable molecular complex with the soluble IL-15 receptor alpha (sIL-15Rα). This heterodimeric IL-15:sIL-15Rα complex (hetIL-15) shows therapeutic potential by promoting the growth, mobilization and activation of lymphocytes and is currently evaluated in clinical trials. Favorable pharmacokinetic properties are associated with the heterodimeric formation and the glycosylation of hetIL-15, which, however, remains largely uncharacterized. We report the site-specific N- and O-glycosylation of two clinically relevant large-scale preparations of HEK293-derived recombinant human hetIL-15. Intact IL-15 and sIL-15Rα and derived glycans and glycopeptides were separately profiled using multiple LC-MS/MS strategies. IL-15 Asn79 and sIL-15Rα Asn107 carried the same repertoire of biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures. The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. Mass analysis of intact IL-15 confirmed its N-glycosylation and suggested that Asn79-glycosylation partially prevents Asn77-deamidation. IL-15 contained no O-glycans, whereas sIL-15Rα was heavily O-glycosylated with partially sialylated core 1 and 2-type mono- to hexasaccharides on Thr2, Thr81, Thr86, Thr156, Ser158, and Ser160. The sialoglycans displayed α2-3- and α2-6-NeuAc-type sialylation. Non-human, potentially immunogenic glycoepitopes (e.g.N-glycolylneuraminic acid and α-galactosylation) were not displayed by hetIL-15. Highly reproducible glycosylation of IL-15 and sIL-15Rα of two batches of hetIL-15 demonstrated consistent manufacturing and purification. In conclusion, we document the heterogeneous and reproducible N- and O-glycosylation of large-scale preparations of the therapeutic candidate hetIL-15. Site-specific mapping of these molecular features is important to evaluate the consistent large-scale production and clinical efficacy of hetIL-15.
dc.description.peerreviewedYes
dc.languageEnglish
dc.language.isoeng
dc.publisherSpringer
dc.relation.ispartofpagefrom417
dc.relation.ispartofpageto433
dc.relation.ispartofissue3
dc.relation.ispartofjournalGlycoconjugate Journal
dc.relation.ispartofvolume33
dc.subject.fieldofresearchBiochemistry and Cell Biology not elsewhere classified
dc.subject.fieldofresearchBiochemistry and Cell Biology
dc.subject.fieldofresearchMedical Microbiology
dc.subject.fieldofresearchNeurosciences
dc.subject.fieldofresearchcode060199
dc.subject.fieldofresearchcode0601
dc.subject.fieldofresearchcode1108
dc.subject.fieldofresearchcode1109
dc.titleRecombinant human heterodimeric IL-15 complex displays extensive and reproducible N- and O-linked glycosylation
dc.typeJournal article
dc.type.descriptionC1 - Articles
dc.type.codeC - Journal Articles
gro.hasfulltextNo Full Text
gro.griffith.authorPacker, Nicki


Files in this item

FilesSizeFormatView

There are no files associated with this item.

This item appears in the following Collection(s)

  • Journal articles
    Contains articles published by Griffith authors in scholarly journals.

Show simple item record