Stable expression of a human-like sialylated recombinant thyrotropin in a Chinese hamster ovary cell line expressing α2,6-sialyltransferase

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dc.contributor.authorDAMIANI, RENATApt_BR
dc.contributor.authorOLIVEIRA, JOAO E.pt_BR
dc.contributor.authorVORAUER-UHL, KAROLApt_BR
dc.contributor.authorPERONI, CIBELE N.pt_BR
dc.contributor.authorVIANNA, ELIZABETH G.pt_BR
dc.contributor.authorBARTOLINI, PAOLOpt_BR
dc.contributor.authorRIBELA, MARIA T.C.P.pt_BR
dc.coverageInternacionalpt_BR
dc.date.accessioned2014-07-15T13:41:33Zpt_BR
dc.date.accessioned2014-07-30T11:51:54Z
dc.date.available2014-07-15T13:41:33Zpt_BR
dc.date.available2014-07-30T11:51:54Z
dc.date.issued2009pt_BR
dc.description.abstractA CHO cell line, previously genetically modified by the introduction of rat α2,6-sialyltransferase cDNA, generated for the first time a human-like sialylated recombinant hTSH (hlsr-hTSH) more similar to the native hormone, with 61% of α2,3- and 39% of α2,6-linked sialic acid residues. The best clone, when submitted to gene amplification with up to 8 μM methotrexate, presented a secretion level of ∼2 μg hTSH/106 cells/day, useful for product purification and characterization. The relative molecular masses (Mr) of the heterodimer and of the α- and β-subunits of purified hlsr-hTSH, determined by MALDI-TOF mass spectrometry, and the relative hydrophobicities, determined by RP-HPLC, were not remarkably different from those presented by two r-hTSH preparations secreted by normal CHO cells. Some differences were observed, though, in N-glycan composition, with more tri- and much more tetra-sialylated structures in hlsr-hTSH. When analyzed via an in vivo bioassay based on hTSH-induced T4 release in mice, hlsr-hTSH was shown to be equipotent (p > 0.05) with the commercial preparation of r-hTSH (Thyrogen), and 1.6-fold more potent than native hTSH (p < 0.001).
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)pt_BR
dc.description.sponsorshipIDFAPESP:07/56094-2pt_BR
dc.format.extent7-14pt_BR
dc.identifier.citationDAMIANI, RENATA; OLIVEIRA, JOAO E.; VORAUER-UHL, KAROLA; PERONI, CIBELE N.; VIANNA, ELIZABETH G.; BARTOLINI, PAOLO; RIBELA, MARIA T.C.P. Stable expression of a human-like sialylated recombinant thyrotropin in a Chinese hamster ovary cell line expressing α2,6-sialyltransferase. <b>Protein Expression and Purification</b>, v. 67, n. 1, p. 7-14, 2009. DOI: <a href="https://dx.doi.org/10.1016/j.pep.2009.04.005">10.1016/j.pep.2009.04.005</a>. Disponível em: http://repositorio.ipen.br/handle/123456789/4850.
dc.identifier.doi10.1016/j.pep.2009.04.005
dc.identifier.fasciculo1pt_BR
dc.identifier.issn1046-5928pt_BR
dc.identifier.orcidhttps://orcid.org/0000-0001-8194-5230
dc.identifier.urihttp://repositorio.ipen.br/handle/123456789/4850pt_BR
dc.identifier.vol67pt_BR
dc.relation.ispartofProtein Expression and Purificationpt_BR
dc.rightsopenAccessen
dc.subjectcho cellspt_BR
dc.subjecttshpt_BR
dc.subjecttrhpt_BR
dc.subjection exchange chromatographypt_BR
dc.subjecthigh-performance liquid chromatographypt_BR
dc.subjectbioassaypt_BR
dc.titleStable expression of a human-like sialylated recombinant thyrotropin in a Chinese hamster ovary cell line expressing α2,6-sialyltransferasept_BR
dc.typeArtigo de periódicopt_BR
dspace.entity.typePublication
ipen.autorMARIA TERESA DE CARVALHO PINTO RIBELA
ipen.autorPAOLO BARTOLINI
ipen.autorELIZABETH KINUYO GIMBO VIANNA
ipen.autorCIBELE NUNES PERONI
ipen.autorJOAO EZEQUIEL DE OLIVEIRA
ipen.autorRENATA DAMIANI
ipen.codigoautor1197
ipen.codigoautor1503
ipen.codigoautor510
ipen.codigoautor947
ipen.codigoautor425
ipen.codigoautor3931
ipen.contributor.ipenauthorMARIA TERESA DE CARVALHO PINTO RIBELA
ipen.contributor.ipenauthorPAOLO BARTOLINI
ipen.contributor.ipenauthorELIZABETH KINUYO GIMBO VIANNA
ipen.contributor.ipenauthorCIBELE NUNES PERONI
ipen.contributor.ipenauthorJOAO EZEQUIEL DE OLIVEIRA
ipen.contributor.ipenauthorRENATA DAMIANI
ipen.date.recebimento09-12pt_BR
ipen.identifier.fi1.563pt_BR
ipen.identifier.ipendoc14267pt_BR
ipen.identifier.iwosWoSpt_BR
ipen.range.fi1.500 - 2.999
ipen.type.genreArtigo
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sigepi.autor.atividadeDAMIANI, RENATA:3931:810:Spt_BR
sigepi.autor.atividadeOLIVEIRA, JOÃO E.:425:810:Npt_BR
sigepi.autor.atividadePERONI, CIBELE N.:947:810:Npt_BR
sigepi.autor.atividadeVIANNA, ELIZABETH G.:510:-1:Npt_BR
sigepi.autor.atividadeBARTOLINI, PAOLO:1503:810:Npt_BR
sigepi.autor.atividadeRIBELA, MARIA T.C.P.:1197:810:Npt_BR
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