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

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dc.contributor.authorSuzuki, Miriam Fussae
dc.contributor.authorOliveira, João Ezequiel
dc.contributor.authorDamiani, Renata
dc.contributor.authorLima, Eliana Rosa
dc.contributor.authorAmaral, Kleicy Cavalcante
dc.contributor.authorSantos, Anderson Maikon de Souza [UNESP]
dc.contributor.authorMagalhães, Geraldo Santana
dc.contributor.authorFaverani, Leonardo Perez [UNESP]
dc.contributor.authorPereira, Luis Antonio Violin Dias
dc.contributor.authorSilva, Fabiana Medeiros
dc.contributor.authorBartolini, Paolo
dc.contributor.institutionIPEN-CNEN/SP
dc.contributor.institutionBiosintesis P&D
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.contributor.institutionInstituto Butantan
dc.contributor.institutionUniversidade Estadual de Campinas (UNICAMP)
dc.date.accessioned2020-12-12T01:15:49Z
dc.date.available2020-12-12T01:15:49Z
dc.date.issued2020-12-01
dc.description.abstractCommercially available preparations of methionyl-human BMP-2 and CHO-derived hBMP-2, which belongs to the transforming growth factor β (TGF-β) 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.en
dc.description.affiliationBiotechnology Center Instituto de Pesquisas Energéticas e Nucleares IPEN-CNEN/SP, Avenida Prof. Lineu Prestes 2242, Cidade Universitária
dc.description.affiliationBiosintesis P&D
dc.description.affiliationDepartment of Surgery and Integrated Clinic-Universidade Estadual Paulista Júlio de Mesquita Filho-UNESP School of Dentistry
dc.description.affiliationImmunopathology Laboratory Instituto Butantan
dc.description.affiliationDepartment of Biochemistry and Tissue Biology Institute of Biology Universidade Estadual de Campinas-UNICAMP
dc.description.affiliationUnespDepartment of Surgery and Integrated Clinic-Universidade Estadual Paulista Júlio de Mesquita Filho-UNESP School of Dentistry
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipIdFAPESP: 2015/15446-0
dc.description.sponsorshipIdFAPESP: 2016/24724-6
dc.identifierhttp://dx.doi.org/10.1186/s13568-020-0964-5
dc.identifier.citationAMB Express, v. 10, n. 1, 2020.
dc.identifier.doi10.1186/s13568-020-0964-5
dc.identifier.issn2191-0855
dc.identifier.scopus2-s2.0-85079699034
dc.identifier.urihttp://hdl.handle.net/11449/198547
dc.language.isoeng
dc.relation.ispartofAMB Express
dc.sourceScopus
dc.subjectBMP-2
dc.subjectC2C12 bioassay
dc.subjectCHO cell-derived
dc.subjectEfficient osteoinductor
dc.subjectEscherichia coli-derived
dc.titleHuman bone morphogenetic protein-2 (hBMP-2) characterization by physical–chemical, immunological and biological assaysen
dc.typeArtigo
unesp.author.orcid0000-0002-7467-3457[1]
unesp.author.orcid0000-0002-6937-1120[2]
unesp.author.orcid0000-0002-8314-0230[3]
unesp.author.orcid0000-0002-2205-2587[4]
unesp.author.orcid0000-0002-5174-4857[5]
unesp.author.orcid0000-0001-9371-9417[6]
unesp.author.orcid0000-0003-4292-7827[7]
unesp.author.orcid0000-0003-2249-3048[8]
unesp.author.orcid0000-0002-9332-7285[9]
unesp.author.orcid0000-0001-8232-644X[10]
unesp.author.orcid0000-0002-5258-1527[11]

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