Human bone morphogenetic protein-2 (hBMP-2) characterization by physical–chemical, immunological and biological assays

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dc.contributorLab. Imunopatologiapt_BR
dc.contributor.authorSuzuki, Miriam Fussaept_BR
dc.contributor.authorOliveira, João Ezequielpt_BR
dc.contributor.authorDamiani, Renatapt_BR
dc.contributor.authorLima, Eliana Rosapt_BR
dc.contributor.authorAmaral, Kleicy Cavalcantept_BR
dc.contributor.authorSantos, Anderson Maikon de Souzapt_BR
dc.contributor.authorMagalhães, Geraldo Santanapt_BR
dc.contributor.authorFaverani, Leonardo Perezpt_BR
dc.contributor.authorPereira, Luis Antonio Violin Diaspt_BR
dc.contributor.authorSilva, Fabiana Medeirospt_BR
dc.contributor.authorBartolini, Paolopt_BR
dc.date.accessioned2020-07-09T21:26:44Z-
dc.date.available2020-07-09T21:26:44Z-
dc.date.issued2020pt_BR
dc.identifier.citationSuzuki MF, Oliveira JE, Damiani R, Lima ER, Amaral KC, Santos AMS, et al. Human bone morphogenetic protein-2 (hBMP-2) characterization by physical-chemical, immunological and biological assays. AMB Express. 2020 Feb;10:34. doi:10.1186/s13568-020-0964-5.pt_BR
dc.identifier.urihttps://repositorio.butantan.gov.br/handle/butantan/2957-
dc.description.abstractCommercially available preparations of methionyl-human BMP-2 and CHO-derived hBMP-2, which belongs to the transforming growth factor ß (TGF-ß) superfamily, were used for a complete characterization. This protein is an extremely efficient osteoinductor that plays an important role during bone regeneration and embryonic development. Characterization was carried out via SDS-PAGE and Western blotting, followed by reversed-phase HPLC, size-exclusion HPLC and MALDI-TOF-MS. The classical in vitro bioassay, based on the induction of alkaline phosphatase activity in C2C12 cells, confirmed that hBMP-2 biological activity is mostly related to the dimeric form, being ~ 4-fold higher for the CHO-derived glycosylated form when compared with the E. coli counterpart. The E. coli-derived met-hBMP-2 has shown, by MALDI-TOF-MS, a large presence of the bioactive dimer. A more complex molecular mass (MM) distribution was found for the CHO-derived product, whose exact MM has never been reported because of its variable glycosylation. A method based on RP-HPLC was set up, allowing a quantitative and qualitative hBMP-2 determination even directly on ongoing culture media. Considering that hBMP-2 is highly unstable, presenting moreover an extremely high aggregate value, we believe that these data pave the way to a necessary characterization of this important factor when synthesized by DNA recombinant techniques in different types of hosts.pt_BR
dc.description.sponsorship(FAPESP) Fundação de Amparo à Pesquisa do Estado de São Paulopt_BR
dc.format.extent34pt_BR
dc.language.isoEnglishpt_BR
dc.relation.ispartofAMB Expresspt_BR
dc.rightsOpen accesspt_BR
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/pt_BR
dc.titleHuman bone morphogenetic protein-2 (hBMP-2) characterization by physical–chemical, immunological and biological assayspt_BR
dc.typeArticlept_BR
dc.rights.licenseCC BYpt_BR
dc.identifier.doi10.1186/s13568-020-0964-5pt_BR
dc.identifier.urlhttps://doi.org/10.1186/s13568-020-0964-5pt_BR
dc.contributor.external(IPEN) Instituto de Pesquisas Energéticas e Nuclearespt_BR
dc.contributor.externalBiosintesis P&Dpt_BR
dc.contributor.external(UNESP) Universidade Estadual Paulista Júlio de Mesquita Filhopt_BR
dc.contributor.external(UNICAMP) Universidade Estadual de Campinaspt_BR
dc.identifier.citationvolume10pt_BR
dc.subject.keywordBMP-2pt_BR
dc.subject.keywordEscherichia coli-derivedpt_BR
dc.subject.keywordCHO cell-derivedpt_BR
dc.subject.keywordC2C12 bioassaypt_BR
dc.subject.keywordEfficient osteoinductorpt_BR
dc.relation.ispartofabbreviatedAMB Expresspt_BR
dc.identifier.citationabntv. 10, 34, fev. 2020pt_BR
dc.identifier.citationvancouver2020 Feb;10:34pt_BR
dc.contributor.butantanMagalhães, Geraldo Santana|:Pesquisador:Docente Permanente PPGTOX|:Lab. Imunopatologia|:pt_BR
dc.sponsorship.butantanFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)¦¦2015/15446-0pt_BR
dc.sponsorship.butantanFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)¦¦2016/24724-6pt_BR
dc.identifier.bvsccBR78.1pt_BR
dc.identifier.bvsdbIBProdpt_BR
dc.description.dbindexedYespt_BR
item.grantfulltextopen-
item.languageiso639-1English-
item.fulltextCom Texto completo-
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