Recombinant human heterodimeric IL-15 complex displays extensive and reproducible N- and O-linked glycosylation View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2015-11-12

AUTHORS

M. Thaysen-Andersen, E. Chertova, C. Bergamaschi, E. S. X. Moh, O. Chertov, J. Roser, R. Sowder, J. Bear, J. Lifson, N. H. Packer, B. K. Felber, G. N. Pavlakis

ABSTRACT

Human 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. More... »

PAGES

417-433

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s10719-015-9627-1

DOI

http://dx.doi.org/10.1007/s10719-015-9627-1

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1025232579

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/26563299


Indexing Status Check whether this publication has been indexed by Scopus and Web Of Science using the SN Indexing Status Tool
Incoming Citations Browse incoming citations for this publication using opencitations.net

JSON-LD is the canonical representation for SciGraph data.

TIP: You can open this SciGraph record using an external JSON-LD service: JSON-LD Playground Google SDTT

[
  {
    "@context": "https://springernature.github.io/scigraph/jsonld/sgcontext.json", 
    "about": [
      {
        "id": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/06", 
        "inDefinedTermSet": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/", 
        "name": "Biological Sciences", 
        "type": "DefinedTerm"
      }, 
      {
        "id": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/0601", 
        "inDefinedTermSet": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/", 
        "name": "Biochemistry and Cell Biology", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Acetylglucosamine", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Glycosylation", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "HEK293 Cells", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Humans", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Interleukin-15", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Protein Binding", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Protein Processing, Post-Translational", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Receptors, Interleukin-15", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Recombinant Proteins", 
        "type": "DefinedTerm"
      }
    ], 
    "author": [
      {
        "affiliation": {
          "alternateName": "Department of Chemistry and Biomolecular Sciences, Macquarie University, 2109, Sydney, NSW, Australia", 
          "id": "http://www.grid.ac/institutes/grid.1004.5", 
          "name": [
            "Department of Chemistry and Biomolecular Sciences, Macquarie University, 2109, Sydney, NSW, Australia"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Thaysen-Andersen", 
        "givenName": "M.", 
        "id": "sg:person.01164757334.00", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01164757334.00"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "AIDS and Cancer Virus Program, Leidos Biomedical Research, Inc., Frederick National Laboratory, 21702, Frederick, MD, USA", 
          "id": "http://www.grid.ac/institutes/grid.418021.e", 
          "name": [
            "AIDS and Cancer Virus Program, Leidos Biomedical Research, Inc., Frederick National Laboratory, 21702, Frederick, MD, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Chertova", 
        "givenName": "E.", 
        "id": "sg:person.01227343507.09", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01227343507.09"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Human Retrovirus Pathogenesis Section, Vaccine Branch, Center for Cancer Research, National Cancer Institute at Frederick, 21702, Frederick, MD, USA", 
          "id": "http://www.grid.ac/institutes/grid.417768.b", 
          "name": [
            "Human Retrovirus Section, Vaccine Branch, Center for Cancer Research, National Cancer Institute at Frederick, 21702, Frederick, MD, USA", 
            "Human Retrovirus Pathogenesis Section, Vaccine Branch, Center for Cancer Research, National Cancer Institute at Frederick, 21702, Frederick, MD, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Bergamaschi", 
        "givenName": "C.", 
        "id": "sg:person.0703560446.90", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0703560446.90"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Department of Chemistry and Biomolecular Sciences, Macquarie University, 2109, Sydney, NSW, Australia", 
          "id": "http://www.grid.ac/institutes/grid.1004.5", 
          "name": [
            "Department of Chemistry and Biomolecular Sciences, Macquarie University, 2109, Sydney, NSW, Australia"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Moh", 
        "givenName": "E. S. X.", 
        "id": "sg:person.0661077670.29", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0661077670.29"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Cancer Research Technology Program, Leidos Biomedical, Inc., Frederick National Laboratory, 21702, Frederick, MD, USA", 
          "id": "http://www.grid.ac/institutes/grid.418021.e", 
          "name": [
            "Cancer Research Technology Program, Leidos Biomedical, Inc., Frederick National Laboratory, 21702, Frederick, MD, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Chertov", 
        "givenName": "O.", 
        "id": "sg:person.01161230307.91", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01161230307.91"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "AIDS and Cancer Virus Program, Leidos Biomedical Research, Inc., Frederick National Laboratory, 21702, Frederick, MD, USA", 
          "id": "http://www.grid.ac/institutes/grid.418021.e", 
          "name": [
            "AIDS and Cancer Virus Program, Leidos Biomedical Research, Inc., Frederick National Laboratory, 21702, Frederick, MD, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Roser", 
        "givenName": "J.", 
        "id": "sg:person.01011355020.84", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01011355020.84"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "AIDS and Cancer Virus Program, Leidos Biomedical Research, Inc., Frederick National Laboratory, 21702, Frederick, MD, USA", 
          "id": "http://www.grid.ac/institutes/grid.418021.e", 
          "name": [
            "AIDS and Cancer Virus Program, Leidos Biomedical Research, Inc., Frederick National Laboratory, 21702, Frederick, MD, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Sowder", 
        "givenName": "R.", 
        "id": "sg:person.01056275331.50", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01056275331.50"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Human Retrovirus Pathogenesis Section, Vaccine Branch, Center for Cancer Research, National Cancer Institute at Frederick, 21702, Frederick, MD, USA", 
          "id": "http://www.grid.ac/institutes/grid.417768.b", 
          "name": [
            "Human Retrovirus Pathogenesis Section, Vaccine Branch, Center for Cancer Research, National Cancer Institute at Frederick, 21702, Frederick, MD, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Bear", 
        "givenName": "J.", 
        "id": "sg:person.01167147567.45", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01167147567.45"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "AIDS and Cancer Virus Program, Leidos Biomedical Research, Inc., Frederick National Laboratory, 21702, Frederick, MD, USA", 
          "id": "http://www.grid.ac/institutes/grid.418021.e", 
          "name": [
            "AIDS and Cancer Virus Program, Leidos Biomedical Research, Inc., Frederick National Laboratory, 21702, Frederick, MD, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Lifson", 
        "givenName": "J.", 
        "id": "sg:person.011472222277.74", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.011472222277.74"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Department of Chemistry and Biomolecular Sciences, Macquarie University, 2109, Sydney, NSW, Australia", 
          "id": "http://www.grid.ac/institutes/grid.1004.5", 
          "name": [
            "Department of Chemistry and Biomolecular Sciences, Macquarie University, 2109, Sydney, NSW, Australia"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Packer", 
        "givenName": "N. H.", 
        "id": "sg:person.0657131714.23", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0657131714.23"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Human Retrovirus Pathogenesis Section, Vaccine Branch, Center for Cancer Research, National Cancer Institute at Frederick, 21702, Frederick, MD, USA", 
          "id": "http://www.grid.ac/institutes/grid.417768.b", 
          "name": [
            "Human Retrovirus Pathogenesis Section, Vaccine Branch, Center for Cancer Research, National Cancer Institute at Frederick, 21702, Frederick, MD, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Felber", 
        "givenName": "B. K.", 
        "id": "sg:person.01364712446.92", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01364712446.92"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Human Retrovirus Section, Vaccine Branch, Center for Cancer Research, National Cancer Institute at Frederick, 21702, Frederick, MD, USA", 
          "id": "http://www.grid.ac/institutes/grid.417768.b", 
          "name": [
            "Human Retrovirus Section, Vaccine Branch, Center for Cancer Research, National Cancer Institute at Frederick, 21702, Frederick, MD, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Pavlakis", 
        "givenName": "G. N.", 
        "id": "sg:person.01134012117.85", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01134012117.85"
        ], 
        "type": "Person"
      }
    ], 
    "citation": [
      {
        "id": "sg:pub.10.1038/gt.2014.84", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1038475105", 
          "https://doi.org/10.1038/gt.2014.84"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1038/nbt1110-1153", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1014137378", 
          "https://doi.org/10.1038/nbt1110-1153"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1038/ni.2449", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1003634516", 
          "https://doi.org/10.1038/ni.2449"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1007/s11095-011-0597-0", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1033156504", 
          "https://doi.org/10.1007/s11095-011-0597-0"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1038/nprot.2012.063", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1035692290", 
          "https://doi.org/10.1038/nprot.2012.063"
        ], 
        "type": "CreativeWork"
      }
    ], 
    "datePublished": "2015-11-12", 
    "datePublishedReg": "2015-11-12", 
    "description": "Human interleukin 15 (IL-15) circulates in blood as a stable molecular complex with the soluble IL-15 receptor alpha (sIL-15R\u03b1). This heterodimeric IL-15:sIL-15R\u03b1 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-15R\u03b1 and derived glycans and glycopeptides were separately profiled using multiple LC-MS/MS strategies. IL-15 Asn79 and sIL-15R\u03b1 Asn107 carried the same repertoire of biosynthetically-related N-glycans covering mostly \u03b11-6-core-fucosylated and \u03b2-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-15R\u03b1 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 \u03b12-3- and \u03b12-6-NeuAc-type sialylation. Non-human, potentially immunogenic glycoepitopes (e.g.N-glycolylneuraminic acid and \u03b1-galactosylation) were not displayed by hetIL-15. Highly reproducible glycosylation of IL-15 and sIL-15R\u03b1 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.", 
    "genre": "article", 
    "id": "sg:pub.10.1007/s10719-015-9627-1", 
    "inLanguage": "en", 
    "isAccessibleForFree": false, 
    "isFundedItemOf": [
      {
        "id": "sg:grant.2710380", 
        "type": "MonetaryGrant"
      }, 
      {
        "id": "sg:grant.2723821", 
        "type": "MonetaryGrant"
      }, 
      {
        "id": "sg:grant.2723653", 
        "type": "MonetaryGrant"
      }, 
      {
        "id": "sg:grant.2343593", 
        "type": "MonetaryGrant"
      }
    ], 
    "isPartOf": [
      {
        "id": "sg:journal.1096094", 
        "issn": [
          "0969-3653", 
          "1573-4986"
        ], 
        "name": "Glycoconjugate Journal", 
        "publisher": "Springer Nature", 
        "type": "Periodical"
      }, 
      {
        "issueNumber": "3", 
        "type": "PublicationIssue"
      }, 
      {
        "type": "PublicationVolume", 
        "volumeNumber": "33"
      }
    ], 
    "keywords": [
      "site-specific mapping", 
      "complex-type structures", 
      "LC-MS/MS strategy", 
      "glycosylation sites", 
      "heterodimeric formation", 
      "glycosylation", 
      "receptor interface", 
      "glycans", 
      "same repertoire", 
      "core 1", 
      "large-scale production", 
      "MS strategy", 
      "receptor alpha", 
      "IL-15 receptor alpha", 
      "molecular features", 
      "large-scale preparation", 
      "molecular complexes", 
      "activation of lymphocytes", 
      "Thr156", 
      "Ser158", 
      "therapeutic potential", 
      "favorable pharmacokinetic properties", 
      "complexes", 
      "clinical efficacy", 
      "Ser160", 
      "interleukin 15", 
      "clinical trials", 
      "glycoepitopes", 
      "IL-15", 
      "Thr81", 
      "heterodimeric IL-15", 
      "Human interleukin 15", 
      "Asn79", 
      "pharmacokinetic properties", 
      "Thr2", 
      "sialoglycans", 
      "complex displays", 
      "purification", 
      "sialylation", 
      "activation", 
      "repertoire", 
      "Thr86", 
      "growth", 
      "mass analysis", 
      "stable molecular complexes", 
      "sites", 
      "\u03b11", 
      "alpha", 
      "glycopeptides", 
      "production", 
      "hexasaccharide", 
      "lymphocytes", 
      "\u03b12", 
      "mapping", 
      "blood", 
      "trials", 
      "formation", 
      "consistent manufacturing", 
      "efficacy", 
      "mobilization", 
      "conclusion", 
      "potential", 
      "structure", 
      "mono", 
      "analysis", 
      "strategies", 
      "preparation", 
      "display", 
      "features", 
      "batch", 
      "properties", 
      "interface", 
      "manufacturing", 
      "soluble IL-15 receptor alpha", 
      "sIL-15R\u03b1 complex", 
      "glycosylation of hetIL-15", 
      "hetIL-15", 
      "relevant large-scale preparations", 
      "HEK293-derived recombinant human hetIL-15", 
      "recombinant human hetIL-15", 
      "human hetIL-15", 
      "Intact IL-15", 
      "sIL-15R\u03b1", 
      "multiple LC-MS/MS strategies", 
      "IL-15 Asn79", 
      "sIL-15R\u03b1 Asn107", 
      "Asn107", 
      "GlcNAc-terminating complex-type structures", 
      "potential IL-15 N", 
      "IL-15 N", 
      "IL-2 receptor interface", 
      "Asn79-glycosylation", 
      "Asn77-deamidation", 
      "\u03b12-6-NeuAc", 
      "type sialylation", 
      "immunogenic glycoepitopes", 
      "reproducible glycosylation", 
      "batches of hetIL-15", 
      "therapeutic candidate hetIL-15", 
      "candidate hetIL-15", 
      "consistent large-scale production", 
      "Recombinant human heterodimeric IL-15 complex displays", 
      "human heterodimeric IL-15 complex displays", 
      "heterodimeric IL-15 complex displays", 
      "IL-15 complex displays"
    ], 
    "name": "Recombinant human heterodimeric IL-15 complex displays extensive and reproducible N- and O-linked glycosylation", 
    "pagination": "417-433", 
    "productId": [
      {
        "name": "dimensions_id", 
        "type": "PropertyValue", 
        "value": [
          "pub.1025232579"
        ]
      }, 
      {
        "name": "doi", 
        "type": "PropertyValue", 
        "value": [
          "10.1007/s10719-015-9627-1"
        ]
      }, 
      {
        "name": "pubmed_id", 
        "type": "PropertyValue", 
        "value": [
          "26563299"
        ]
      }
    ], 
    "sameAs": [
      "https://doi.org/10.1007/s10719-015-9627-1", 
      "https://app.dimensions.ai/details/publication/pub.1025232579"
    ], 
    "sdDataset": "articles", 
    "sdDatePublished": "2021-12-01T19:32", 
    "sdLicense": "https://scigraph.springernature.com/explorer/license/", 
    "sdPublisher": {
      "name": "Springer Nature - SN SciGraph project", 
      "type": "Organization"
    }, 
    "sdSource": "s3://com-springernature-scigraph/baseset/20211201/entities/gbq_results/article/article_651.jsonl", 
    "type": "ScholarlyArticle", 
    "url": "https://doi.org/10.1007/s10719-015-9627-1"
  }
]
 

Download the RDF metadata as:  json-ld nt turtle xml License info

HOW TO GET THIS DATA PROGRAMMATICALLY:

JSON-LD is a popular format for linked data which is fully compatible with JSON.

curl -H 'Accept: application/ld+json' 'https://scigraph.springernature.com/pub.10.1007/s10719-015-9627-1'

N-Triples is a line-based linked data format ideal for batch operations.

curl -H 'Accept: application/n-triples' 'https://scigraph.springernature.com/pub.10.1007/s10719-015-9627-1'

Turtle is a human-readable linked data format.

curl -H 'Accept: text/turtle' 'https://scigraph.springernature.com/pub.10.1007/s10719-015-9627-1'

RDF/XML is a standard XML format for linked data.

curl -H 'Accept: application/rdf+xml' 'https://scigraph.springernature.com/pub.10.1007/s10719-015-9627-1'


 

This table displays all metadata directly associated to this object as RDF triples.

318 TRIPLES      22 PREDICATES      145 URIs      132 LITERALS      16 BLANK NODES

Subject Predicate Object
1 sg:pub.10.1007/s10719-015-9627-1 schema:about N20c2856c734746e4b4083e0970cce825
2 N344364fcee98414fa6ab4e5007de5851
3 N3a2ff525cdf048cba442077524ef7dc4
4 N4beca268a58847cdb9b94ea25b1ecba9
5 N76f4c2c6967c41f7925bf168d91d5943
6 N77dbbe376bde4784b0a179dd626256f9
7 Nab7681efe9ab485ea7c3d1b7587f270a
8 Nbbdfc6b6653943e1ae47697e14dac17e
9 Nd6d551a69f8045ca8bfb77859acff401
10 anzsrc-for:06
11 anzsrc-for:0601
12 schema:author N71a4feec8f9a487f8181b7261489da18
13 schema:citation sg:pub.10.1007/s11095-011-0597-0
14 sg:pub.10.1038/gt.2014.84
15 sg:pub.10.1038/nbt1110-1153
16 sg:pub.10.1038/ni.2449
17 sg:pub.10.1038/nprot.2012.063
18 schema:datePublished 2015-11-12
19 schema:datePublishedReg 2015-11-12
20 schema:description Human 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.
21 schema:genre article
22 schema:inLanguage en
23 schema:isAccessibleForFree false
24 schema:isPartOf N35a32934ab9a4296b168679e403f85c3
25 N9d46c17f6f584d57924fec609a44e793
26 sg:journal.1096094
27 schema:keywords Asn107
28 Asn77-deamidation
29 Asn79
30 Asn79-glycosylation
31 GlcNAc-terminating complex-type structures
32 HEK293-derived recombinant human hetIL-15
33 Human interleukin 15
34 IL-15
35 IL-15 Asn79
36 IL-15 N
37 IL-15 complex displays
38 IL-15 receptor alpha
39 IL-2 receptor interface
40 Intact IL-15
41 LC-MS/MS strategy
42 MS strategy
43 Recombinant human heterodimeric IL-15 complex displays
44 Ser158
45 Ser160
46 Thr156
47 Thr2
48 Thr81
49 Thr86
50 activation
51 activation of lymphocytes
52 alpha
53 analysis
54 batch
55 batches of hetIL-15
56 blood
57 candidate hetIL-15
58 clinical efficacy
59 clinical trials
60 complex displays
61 complex-type structures
62 complexes
63 conclusion
64 consistent large-scale production
65 consistent manufacturing
66 core 1
67 display
68 efficacy
69 favorable pharmacokinetic properties
70 features
71 formation
72 glycans
73 glycoepitopes
74 glycopeptides
75 glycosylation
76 glycosylation of hetIL-15
77 glycosylation sites
78 growth
79 hetIL-15
80 heterodimeric IL-15
81 heterodimeric IL-15 complex displays
82 heterodimeric formation
83 hexasaccharide
84 human hetIL-15
85 human heterodimeric IL-15 complex displays
86 immunogenic glycoepitopes
87 interface
88 interleukin 15
89 large-scale preparation
90 large-scale production
91 lymphocytes
92 manufacturing
93 mapping
94 mass analysis
95 mobilization
96 molecular complexes
97 molecular features
98 mono
99 multiple LC-MS/MS strategies
100 pharmacokinetic properties
101 potential
102 potential IL-15 N
103 preparation
104 production
105 properties
106 purification
107 receptor alpha
108 receptor interface
109 recombinant human hetIL-15
110 relevant large-scale preparations
111 repertoire
112 reproducible glycosylation
113 sIL-15Rα
114 sIL-15Rα Asn107
115 sIL-15Rα complex
116 same repertoire
117 sialoglycans
118 sialylation
119 site-specific mapping
120 sites
121 soluble IL-15 receptor alpha
122 stable molecular complexes
123 strategies
124 structure
125 therapeutic candidate hetIL-15
126 therapeutic potential
127 trials
128 type sialylation
129 α1
130 α2
131 α2-6-NeuAc
132 schema:name Recombinant human heterodimeric IL-15 complex displays extensive and reproducible N- and O-linked glycosylation
133 schema:pagination 417-433
134 schema:productId N0416cd27624b4d1cab2aeb88e8bf8610
135 N17cd7e9fb61a437f8ef4fc4f33a99a59
136 Ne05ed61e63b445f78f368e31129eeee9
137 schema:sameAs https://app.dimensions.ai/details/publication/pub.1025232579
138 https://doi.org/10.1007/s10719-015-9627-1
139 schema:sdDatePublished 2021-12-01T19:32
140 schema:sdLicense https://scigraph.springernature.com/explorer/license/
141 schema:sdPublisher N38ef0cf310804facbc4086dca58c39a7
142 schema:url https://doi.org/10.1007/s10719-015-9627-1
143 sgo:license sg:explorer/license/
144 sgo:sdDataset articles
145 rdf:type schema:ScholarlyArticle
146 N0416cd27624b4d1cab2aeb88e8bf8610 schema:name pubmed_id
147 schema:value 26563299
148 rdf:type schema:PropertyValue
149 N12e48cb14e814a5e900afccdc32d96c2 rdf:first sg:person.0661077670.29
150 rdf:rest N2421a6103e3d4d3385d50a2540112356
151 N17cd7e9fb61a437f8ef4fc4f33a99a59 schema:name doi
152 schema:value 10.1007/s10719-015-9627-1
153 rdf:type schema:PropertyValue
154 N20c2856c734746e4b4083e0970cce825 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
155 schema:name Acetylglucosamine
156 rdf:type schema:DefinedTerm
157 N2421a6103e3d4d3385d50a2540112356 rdf:first sg:person.01161230307.91
158 rdf:rest Ndc15cc360b7a48bf9c3243c414ec5f57
159 N2bbfd74f295c41838a5c001a9fd83a1d rdf:first sg:person.01227343507.09
160 rdf:rest Ne9f7c58059af4898b9f4cd051cd7ee5f
161 N344364fcee98414fa6ab4e5007de5851 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
162 schema:name Protein Processing, Post-Translational
163 rdf:type schema:DefinedTerm
164 N35a32934ab9a4296b168679e403f85c3 schema:volumeNumber 33
165 rdf:type schema:PublicationVolume
166 N38ef0cf310804facbc4086dca58c39a7 schema:name Springer Nature - SN SciGraph project
167 rdf:type schema:Organization
168 N3a2ff525cdf048cba442077524ef7dc4 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
169 schema:name Protein Binding
170 rdf:type schema:DefinedTerm
171 N4beca268a58847cdb9b94ea25b1ecba9 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
172 schema:name HEK293 Cells
173 rdf:type schema:DefinedTerm
174 N4eef70d821294825b8cc135926ceacd0 rdf:first sg:person.01134012117.85
175 rdf:rest rdf:nil
176 N569c222511c74fe6a51c1a7611b50538 rdf:first sg:person.01167147567.45
177 rdf:rest N7263c2ccb99d4a8aae1a71bf3444ca24
178 N5af67627ff174f19b0a4ea90496d02b6 rdf:first sg:person.0657131714.23
179 rdf:rest N902e353fef0542e9908876105123a084
180 N71a4feec8f9a487f8181b7261489da18 rdf:first sg:person.01164757334.00
181 rdf:rest N2bbfd74f295c41838a5c001a9fd83a1d
182 N7263c2ccb99d4a8aae1a71bf3444ca24 rdf:first sg:person.011472222277.74
183 rdf:rest N5af67627ff174f19b0a4ea90496d02b6
184 N76f4c2c6967c41f7925bf168d91d5943 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
185 schema:name Recombinant Proteins
186 rdf:type schema:DefinedTerm
187 N77dbbe376bde4784b0a179dd626256f9 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
188 schema:name Interleukin-15
189 rdf:type schema:DefinedTerm
190 N902e353fef0542e9908876105123a084 rdf:first sg:person.01364712446.92
191 rdf:rest N4eef70d821294825b8cc135926ceacd0
192 N9d46c17f6f584d57924fec609a44e793 schema:issueNumber 3
193 rdf:type schema:PublicationIssue
194 Nab7681efe9ab485ea7c3d1b7587f270a schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
195 schema:name Humans
196 rdf:type schema:DefinedTerm
197 Nbbdfc6b6653943e1ae47697e14dac17e schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
198 schema:name Receptors, Interleukin-15
199 rdf:type schema:DefinedTerm
200 Nd6d551a69f8045ca8bfb77859acff401 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
201 schema:name Glycosylation
202 rdf:type schema:DefinedTerm
203 Ndb46e427e3f4435195eff7daac6d7377 rdf:first sg:person.01056275331.50
204 rdf:rest N569c222511c74fe6a51c1a7611b50538
205 Ndc15cc360b7a48bf9c3243c414ec5f57 rdf:first sg:person.01011355020.84
206 rdf:rest Ndb46e427e3f4435195eff7daac6d7377
207 Ne05ed61e63b445f78f368e31129eeee9 schema:name dimensions_id
208 schema:value pub.1025232579
209 rdf:type schema:PropertyValue
210 Ne9f7c58059af4898b9f4cd051cd7ee5f rdf:first sg:person.0703560446.90
211 rdf:rest N12e48cb14e814a5e900afccdc32d96c2
212 anzsrc-for:06 schema:inDefinedTermSet anzsrc-for:
213 schema:name Biological Sciences
214 rdf:type schema:DefinedTerm
215 anzsrc-for:0601 schema:inDefinedTermSet anzsrc-for:
216 schema:name Biochemistry and Cell Biology
217 rdf:type schema:DefinedTerm
218 sg:grant.2343593 http://pending.schema.org/fundedItem sg:pub.10.1007/s10719-015-9627-1
219 rdf:type schema:MonetaryGrant
220 sg:grant.2710380 http://pending.schema.org/fundedItem sg:pub.10.1007/s10719-015-9627-1
221 rdf:type schema:MonetaryGrant
222 sg:grant.2723653 http://pending.schema.org/fundedItem sg:pub.10.1007/s10719-015-9627-1
223 rdf:type schema:MonetaryGrant
224 sg:grant.2723821 http://pending.schema.org/fundedItem sg:pub.10.1007/s10719-015-9627-1
225 rdf:type schema:MonetaryGrant
226 sg:journal.1096094 schema:issn 0969-3653
227 1573-4986
228 schema:name Glycoconjugate Journal
229 schema:publisher Springer Nature
230 rdf:type schema:Periodical
231 sg:person.01011355020.84 schema:affiliation grid-institutes:grid.418021.e
232 schema:familyName Roser
233 schema:givenName J.
234 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01011355020.84
235 rdf:type schema:Person
236 sg:person.01056275331.50 schema:affiliation grid-institutes:grid.418021.e
237 schema:familyName Sowder
238 schema:givenName R.
239 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01056275331.50
240 rdf:type schema:Person
241 sg:person.01134012117.85 schema:affiliation grid-institutes:grid.417768.b
242 schema:familyName Pavlakis
243 schema:givenName G. N.
244 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01134012117.85
245 rdf:type schema:Person
246 sg:person.011472222277.74 schema:affiliation grid-institutes:grid.418021.e
247 schema:familyName Lifson
248 schema:givenName J.
249 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.011472222277.74
250 rdf:type schema:Person
251 sg:person.01161230307.91 schema:affiliation grid-institutes:grid.418021.e
252 schema:familyName Chertov
253 schema:givenName O.
254 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01161230307.91
255 rdf:type schema:Person
256 sg:person.01164757334.00 schema:affiliation grid-institutes:grid.1004.5
257 schema:familyName Thaysen-Andersen
258 schema:givenName M.
259 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01164757334.00
260 rdf:type schema:Person
261 sg:person.01167147567.45 schema:affiliation grid-institutes:grid.417768.b
262 schema:familyName Bear
263 schema:givenName J.
264 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01167147567.45
265 rdf:type schema:Person
266 sg:person.01227343507.09 schema:affiliation grid-institutes:grid.418021.e
267 schema:familyName Chertova
268 schema:givenName E.
269 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01227343507.09
270 rdf:type schema:Person
271 sg:person.01364712446.92 schema:affiliation grid-institutes:grid.417768.b
272 schema:familyName Felber
273 schema:givenName B. K.
274 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01364712446.92
275 rdf:type schema:Person
276 sg:person.0657131714.23 schema:affiliation grid-institutes:grid.1004.5
277 schema:familyName Packer
278 schema:givenName N. H.
279 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0657131714.23
280 rdf:type schema:Person
281 sg:person.0661077670.29 schema:affiliation grid-institutes:grid.1004.5
282 schema:familyName Moh
283 schema:givenName E. S. X.
284 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0661077670.29
285 rdf:type schema:Person
286 sg:person.0703560446.90 schema:affiliation grid-institutes:grid.417768.b
287 schema:familyName Bergamaschi
288 schema:givenName C.
289 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0703560446.90
290 rdf:type schema:Person
291 sg:pub.10.1007/s11095-011-0597-0 schema:sameAs https://app.dimensions.ai/details/publication/pub.1033156504
292 https://doi.org/10.1007/s11095-011-0597-0
293 rdf:type schema:CreativeWork
294 sg:pub.10.1038/gt.2014.84 schema:sameAs https://app.dimensions.ai/details/publication/pub.1038475105
295 https://doi.org/10.1038/gt.2014.84
296 rdf:type schema:CreativeWork
297 sg:pub.10.1038/nbt1110-1153 schema:sameAs https://app.dimensions.ai/details/publication/pub.1014137378
298 https://doi.org/10.1038/nbt1110-1153
299 rdf:type schema:CreativeWork
300 sg:pub.10.1038/ni.2449 schema:sameAs https://app.dimensions.ai/details/publication/pub.1003634516
301 https://doi.org/10.1038/ni.2449
302 rdf:type schema:CreativeWork
303 sg:pub.10.1038/nprot.2012.063 schema:sameAs https://app.dimensions.ai/details/publication/pub.1035692290
304 https://doi.org/10.1038/nprot.2012.063
305 rdf:type schema:CreativeWork
306 grid-institutes:grid.1004.5 schema:alternateName Department of Chemistry and Biomolecular Sciences, Macquarie University, 2109, Sydney, NSW, Australia
307 schema:name Department of Chemistry and Biomolecular Sciences, Macquarie University, 2109, Sydney, NSW, Australia
308 rdf:type schema:Organization
309 grid-institutes:grid.417768.b schema:alternateName Human Retrovirus Pathogenesis Section, Vaccine Branch, Center for Cancer Research, National Cancer Institute at Frederick, 21702, Frederick, MD, USA
310 Human Retrovirus Section, Vaccine Branch, Center for Cancer Research, National Cancer Institute at Frederick, 21702, Frederick, MD, USA
311 schema:name Human Retrovirus Pathogenesis Section, Vaccine Branch, Center for Cancer Research, National Cancer Institute at Frederick, 21702, Frederick, MD, USA
312 Human Retrovirus Section, Vaccine Branch, Center for Cancer Research, National Cancer Institute at Frederick, 21702, Frederick, MD, USA
313 rdf:type schema:Organization
314 grid-institutes:grid.418021.e schema:alternateName AIDS and Cancer Virus Program, Leidos Biomedical Research, Inc., Frederick National Laboratory, 21702, Frederick, MD, USA
315 Cancer Research Technology Program, Leidos Biomedical, Inc., Frederick National Laboratory, 21702, Frederick, MD, USA
316 schema:name AIDS and Cancer Virus Program, Leidos Biomedical Research, Inc., Frederick National Laboratory, 21702, Frederick, MD, USA
317 Cancer Research Technology Program, Leidos Biomedical, Inc., Frederick National Laboratory, 21702, Frederick, MD, USA
318 rdf:type schema:Organization
 




Preview window. Press ESC to close (or click here)


...