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Structural characterization and immunogenicity in wild-type and immune tolerant mice of degraded recombinant human interferon Alpha2b

2005-12Journal article Open access
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dc.contributor.authorHermeling, S.
dc.contributor.authorCaetano, Liliana Aranha
dc.contributor.authorDamen, J. M.
dc.contributor.authorSlijper, M.
dc.contributor.authorSchellekens, H.
dc.contributor.authorCrommelin, D. J.
dc.contributor.authorJiskoot, W.
dc.date.accessioned2013-04-23T13:33:08Z
dc.date.available2013-04-23T13:33:08Z
dc.date.issued2005-12
dc.description.abstractPurpose: This study was conducted to study the influence of protein structure on the immunogenicity in wild-type and immune tolerant mice of well-characterized degradation products of recombinant human interferon alpha2b (rhIFNα2b). Methods: RhIFNα2b was degraded by metal-catalyzed oxidation (M), cross-linking with glutaraldehyde (G), oxidation with hydrogen peroxide (H), and incubation in a boiling water bath (B). The products were characterized with UV absorption, circular dichroism and fluorescence spectroscopy, gel permeation chromatography, reverse-phase high-pressure liquid chromatography, sodium dodecyl sulfate polyacrylamide gel electrophoresis, Western blotting, and mass spectrometry. The immunogenicity of the products was evaluated in wild-type mice and in transgenic mice immune tolerant for hIFNα2. Serum antibodies were detected by enzyme-linked immunosorbent assay or surface plasmon resonance. Results: M-rhIFNα2b contained covalently aggregated rhIFNα2b with three methionines partly oxidized to methionine sulfoxides. G-rhIFNα2b contained covalent aggregates and did not show changes in secondary structure. H-rhIFNα2b was only chemically changed with four partly oxidized methionines. B-rhIFNα2b was largely unfolded and heavily aggregated. Nontreated (N) rhIFNα2b was immunogenic in the wild-type mice but not in the transgenic mice, showing that the latter were immune tolerant for rhIFNα2b. The anti-rhIFNα2b antibody levels in the wild-type mice depended on the degradation product: M-rhIFNα2b > H-rhIFNα2b ∼ N-rhIFNα2b ≫ B-rhIFNα2b; G-rhIFNα2b did not induce anti-rhIFNα2b antibodies. In the transgenic mice, only M-rhIFNα2b could break the immune tolerance. Conclusions: RhIFNα2b immunogenicity is related to its structural integrity. Moreover, the immunogenicity of aggregated rhIFNα2b depends on the structure and orientation of the constituent protein molecules and/or on the aggregate size.por
dc.identifier.citationHermeling S, Caetano LA, Damen JM, Slijper M, Schellekens H, Crommelin DJ, et al. Structural characterization and immunogenicity in wild-type and immune tolerant mice of degraded recombinant human interferon Alpha2b. Pharm Res. 2005;22(12):1997-2006.por
dc.identifier.issn1573-904X
dc.identifier.urihttp://hdl.handle.net/10400.21/2453
dc.language.isoengpor
dc.peerreviewedyespor
dc.publisherSpringerpor
dc.relation.publisherversionhttp://link.springer.com/article/10.1007%2Fs11095-005-8177-9por
dc.subjectAnimalspor
dc.subjectBlotting, Westernpor
dc.subjectChromatography, gelpor
dc.subjectChromatography, high pressure liquidpor
dc.subjectCircular dichroismpor
dc.subjectElectrophoresis, polyacrylamide gelpor
dc.subjectEnzyme-linked immunosorbent assaypor
dc.subjectGlutaral/chemistrypor
dc.subjectHumanspor
dc.subjectImmune tolerance/immunologypor
dc.subjectInterferon-alpha/chemistrypor
dc.subjectInterferon-alpha/immunologypor
dc.subjectLightpor
dc.subjectMass spectrometrypor
dc.subjectMetalspor
dc.subjectMicepor
dc.subjectMice, transgenicpor
dc.subjectOxidation-reductionpor
dc.subjectRecombinant proteinspor
dc.subjectScattering, radiationpor
dc.subjectSpectrometry, fluorescencepor
dc.subjectSpectrophotometry, ultravioletpor
dc.subjectSurface plasmon resonancepor
dc.titleStructural characterization and immunogenicity in wild-type and immune tolerant mice of degraded recombinant human interferon Alpha2bpor
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage2006por
oaire.citation.startPage1997por
oaire.citation.titlePharmaceutical Researchpor
oaire.citation.volume22por
rcaap.rightsrestrictedAccesspor
rcaap.typearticlepor

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