Carbohydrate composition and site-occupancy determination in pituitary and recombinant preparations of human thyrotropin

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dc.contributor.authorBARTOLINI, PAOLO
dc.contributor.authorRIBELA, MARIA T.C.P.
dc.contributor.authorDAMIANI, RENATA
dc.contributor.authorSILVA, FELIPE D.
dc.contributor.authorLIMA, ELIANA R.
dc.contributor.authorOLIVEIRA, JOAO E.
dc.contributor.authorPERONI, CIBELE N.
dc.contributor.authorTORJESEN, PETER A.
dc.contributor.authorSOARES, CARLOS R.
dc.coverageInternacionalpt_BR
dc.date.accessioned2018-03-06T13:37:11Z
dc.date.available2018-03-06T13:37:11Z
dc.date.issued2017pt_BR
dc.description.abstractHuman thyrotropin (hTSH) is a glycoprotein with three potential glycosylation sites: two in the -subunit and one in the -subunit. Carbohydrate site-occupancy is frequently neglected in glycoprotein characterization, even if related to folding, trafficking, initiation of inflammation, host defence and congenital disorders of glycosylation (CDG). For the first-time N-glycoprofiling analysis was applied to site-occupancy determination of two native pituitary hTSH, in comparison with three CHO-derived preparations of hTSH, a widely used biopharmaceutical. A single methodology provided: (i) average N-glycan mass; (ii) mass fraction of each monosaccharide and of sulfate; (iii) percent carbohydrate. The results indicate that occupancy (65–87%) and carbohydrate mass (12–19%) can be 34–57% higher in recombinant hormones. The average glycan mass is 24% lower in pituitary hTSH and contains ∼3-fold fewer moles of galactose (P < 0.005) and sialic acid (P < 0.01). The number of moles of fucose per mole of hTSH was found 2.5-fold higher in the pituitary preparations. One of these native preparations, presenting the smallest glycan mass, lowest occupancy, GalNAc, sulfate, Gal and sialic acid contents, also presented the lowest in vivo bioactivity and circulatory half-life. This methodology, extremely important for comparing a recombinant biopharmaceutical to its native equivalent, can be applied to any physiologically or clinical relevant glycoprotein.pt_BR
dc.format.extentS15-S15pt_BR
dc.identifier.citationBARTOLINI, PAOLO; RIBELA, MARIA T.C.P.; DAMIANI, RENATA; SILVA, FELIPE D.; LIMA, ELIANA R.; OLIVEIRA, JOAO E.; PERONI, CIBELE N.; TORJESEN, PETER A.; SOARES, CARLOS R. Carbohydrate composition and site-occupancy determination in pituitary and recombinant preparations of human thyrotropin. <b>Journal of Biotechnology</b>, v. 256, p. S15-S15, 2017. suppl.. DOI: <a href="https://dx.doi.org/10.1016/j.jbiotec.2017.06.053">10.1016/j.jbiotec.2017.06.053</a>. Disponível em: http://repositorio.ipen.br/handle/123456789/28652.
dc.identifier.doi10.1016/j.jbiotec.2017.06.053pt_BR
dc.identifier.issn0168-1656pt_BR
dc.identifier.orcidhttps://orcid.org/0000-0002-7982-1789
dc.identifier.orcidhttps://orcid.org/0000-0001-8194-5230
dc.identifier.suplementosuppl.pt_BR
dc.identifier.urihttp://repositorio.ipen.br/handle/123456789/28652
dc.identifier.vol256pt_BR
dc.relation.ispartofJournal of Biotechnologypt_BR
dc.rightsopenAccesspt_BR
dc.sourceEUROPEAN BIOTECHNOLOGY CONGRESS, May 25-27, 2017, Dubrovnik, Croatiapt_BR
dc.subjectglycoproteins
dc.subjectcarbohydrates
dc.subjecttrh
dc.subjectcongenital diseases
dc.titleCarbohydrate composition and site-occupancy determination in pituitary and recombinant preparations of human thyrotropinpt_BR
dc.typeResumos em periódicospt_BR
dspace.entity.typePublication
ipen.autorFELIPE DOUGLAS DA SILVA
ipen.autorELIANA ROSA LIMA FILHA
ipen.autorCARLOS ROBERTO JORGE SOARES
ipen.autorCIBELE NUNES PERONI
ipen.autorJOAO EZEQUIEL DE OLIVEIRA
ipen.autorRENATA DAMIANI
ipen.autorMARIA TERESA DE CARVALHO PINTO RIBELA
ipen.autorPAOLO BARTOLINI
ipen.codigoautor14230
ipen.codigoautor10276
ipen.codigoautor509
ipen.codigoautor947
ipen.codigoautor425
ipen.codigoautor3931
ipen.codigoautor1197
ipen.codigoautor1503
ipen.contributor.ipenauthorFELIPE DOUGLAS DA SILVA
ipen.contributor.ipenauthorELIANA ROSA LIMA FILHA
ipen.contributor.ipenauthorCARLOS ROBERTO JORGE SOARES
ipen.contributor.ipenauthorCIBELE NUNES PERONI
ipen.contributor.ipenauthorJOAO EZEQUIEL DE OLIVEIRA
ipen.contributor.ipenauthorRENATA DAMIANI
ipen.contributor.ipenauthorMARIA TERESA DE CARVALHO PINTO RIBELA
ipen.contributor.ipenauthorPAOLO BARTOLINI
ipen.date.recebimento18-03pt_BR
ipen.identifier.fi2.599pt_BR
ipen.identifier.ipendoc24397pt_BR
ipen.identifier.iwosWoSpt_BR
ipen.range.fi1.500 - 2.999
ipen.type.genreResumo
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sigepi.autor.atividadeBARTOLINI, PAOLO:1503:810:Spt_BR
sigepi.autor.atividadeRIBELA, MARIA T.C.P.:1197:810:Npt_BR
sigepi.autor.atividadeDAMIANI, RENATA:3931:-1:Npt_BR
sigepi.autor.atividadeSILVA, FELIPE D.:14230:810:Npt_BR
sigepi.autor.atividadeLIMA, ELIANA R.:10276:-1:Npt_BR
sigepi.autor.atividadeOLIVEIRA, JOAO E.:425:810:Npt_BR
sigepi.autor.atividadePERONI, CIBELE N.:947:810:Npt_BR
sigepi.autor.atividadeSOARES, CARLOS R.:509:810:Npt_BR
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