Influence of reduced Co2 environment on the secretion yield, potency and glycan structures of recombinant thyrotropin (hTSH) from CHO cells

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dc.contributor.authorRIBELA, MARIA T.C.P.pt_BR
dc.contributor.authorOLIVEIRA, JOAO E.pt_BR
dc.contributor.authorDAMIANI, RENATApt_BR
dc.contributor.authorCARVALHO, CRISTIANE M.pt_BR
dc.contributor.authorLHOTA, GABRIELEpt_BR
dc.contributor.authorVORAUER-UHL, KAROLApt_BR
dc.contributor.authorBARTOLINI, PAOLOpt_BR
dc.coverageInternacionalpt_BR
dc.creator.eventoPROTEIN EXPRESSION IN ANIMAL CELLS, 8thpt_BR
dc.date.accessioned2014-11-19T10:54:03Zpt_BR
dc.date.accessioned2014-11-19T14:05:18Zpt_BR
dc.date.accessioned2015-04-01T12:45:49Z
dc.date.available2014-11-19T10:54:03Zpt_BR
dc.date.available2014-11-19T14:05:18Zpt_BR
dc.date.available2015-04-01T12:45:49Z
dc.date.eventoSept. 16-20, 2007pt_BR
dc.description.abstractA consistent increase of — 60%, in the secretion yield of CHO-derived hTSH was observed by changing cell 'culture CO2 conditions from 5% CO2 to air enviroment (0.03 % CO2). The quality of the product obtained under both conditions was analysed for what concerns N-glycan structures, charge isomers and biological activity in comparison with a well known commercial preparation (Thyrogen). The N-glycans identified in the three preparations were of the complex type, presenting di-, tri- and tetraantennary structures, with variable level of sialylation. Considering the latter characteristic, which is directly related to in vivo bioactivity, the three preparations have practically an identical percentage (86-88%) of sialylated structures, with some difference in percentage of di- and tii- sialylated glycan. Monosialylated glycans (N2G2S1, N2G1S1 and N2G2S1F) represent the three most abundant structures with 68-69% of all identified forms in the three preparations. The main difference was found in terms of antennarity with 8-10% more N2 structures for hTSH-IPEN produced in the absence of CO2 (-0O2) and 7-9 % more N3 structures for hTSH-IPEN (+CO2) and Thyrogen. Also for what concerns the total percentage of neutral glycans (12- 14 %), the three preparations are practically identical. No remarkable difference in charge isomers was also observed between the three preparations, the isoelectric focusing (IEF) profiles showing six well visible bands in the 5.39 - 7.35 pI range, the three major bands focusing at pI 5.85, 6.20 and 6.63. A considerably different distribution, with more acidic forms was observed, though, for two native pituitary preparations of hTSH. When analyzed via a simple and precise single-dose bioassay, a slightly significant difference (p<0.02) in activity was found between the two IPEN preparations. hTSH-IPEN (+ CO2) potency was non significantly different from that of Thyrogen, both being 1.6-1.8-fold more potent than the native pituitary reference preparation. We can conclude that, at least in the case of CHO-derived hTSH, different production processes may not greatly affect its glycan structures, charge isomer distribution or biological activity.
dc.identifier.citationRIBELA, MARIA T.C.P.; OLIVEIRA, JOAO E.; DAMIANI, RENATA; CARVALHO, CRISTIANE M.; LHOTA, GABRIELE; VORAUER-UHL, KAROLA; BARTOLINI, PAOLO. Influence of reduced Co2 environment on the secretion yield, potency and glycan structures of recombinant thyrotropin (hTSH) from CHO cells. In: PROTEIN EXPRESSION IN ANIMAL CELLS, 8th, Sept. 16-20, 2007, Angra dos Reis, RJ. <b>Abstracts...</b> Disponível em: http://repositorio.ipen.br/handle/123456789/20105.
dc.identifier.urihttp://repositorio.ipen.br/handle/123456789/20105pt_BR
dc.local.eventoAngra dos Reis, RJpt_BR
dc.rightsopenAccesspt_BR
dc.subjectcho cellspt_BR
dc.subjectrecombinant dnapt_BR
dc.subjectthyroxinept_BR
dc.subjecttshpt_BR
dc.subjectsample preparationpt_BR
dc.titleInfluence of reduced Co2 environment on the secretion yield, potency and glycan structures of recombinant thyrotropin (hTSH) from CHO cellspt_BR
dc.typeResumo de eventos científicospt_BR
dspace.entity.typePublication
ipen.autorPAOLO BARTOLINI
ipen.autorCRISTIANE MOREIRA DE CARVALHO
ipen.autorRENATA DAMIANI
ipen.autorJOAO EZEQUIEL DE OLIVEIRA
ipen.autorMARIA TERESA DE CARVALHO PINTO RIBELA
ipen.codigoautor1503
ipen.codigoautor4016
ipen.codigoautor3931
ipen.codigoautor425
ipen.codigoautor1197
ipen.contributor.ipenauthorPAOLO BARTOLINI
ipen.contributor.ipenauthorCRISTIANE MOREIRA DE CARVALHO
ipen.contributor.ipenauthorRENATA DAMIANI
ipen.contributor.ipenauthorJOAO EZEQUIEL DE OLIVEIRA
ipen.contributor.ipenauthorMARIA TERESA DE CARVALHO PINTO RIBELA
ipen.date.recebimento08-05pt_BR
ipen.event.datapadronizada2007pt_BR
ipen.identifier.ipendoc12453pt_BR
ipen.notas.internasAbstractspt_BR
ipen.type.genreResumo
relation.isAuthorOfPublication7d228133-8477-43fb-941d-2cfb6a48c46c
relation.isAuthorOfPublicationc358cfe5-abea-4abc-9224-5cac9a4f0fd1
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relation.isAuthorOfPublicatione9e6346c-9b8b-4d39-8bdd-505ea6997646
relation.isAuthorOfPublication2c1f5cfd-da46-4b49-b209-77aadfb388f1
relation.isAuthorOfPublication.latestForDiscovery2c1f5cfd-da46-4b49-b209-77aadfb388f1
sigepi.autor.atividadeRIBELA, MARIA T.C.P.:1197:18:Spt_BR
sigepi.autor.atividadeOLIVEIRA, JOAO E.:425:18:Npt_BR
sigepi.autor.atividadeDAMIANI, RENATA:3931:18:Npt_BR
sigepi.autor.atividadeCARVALHO, CRISTIANE M.:4016:18:Npt_BR
sigepi.autor.atividadeBARTOLINI, PAOLO:1503:18:Npt_BR
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