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

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Autor(es): dc.contributorIPEN-CNEN/SP-
Autor(es): dc.contributorBiosintesis P&D-
Autor(es): dc.contributorUniversidade Estadual Paulista (Unesp)-
Autor(es): dc.contributorInstituto Butantan-
Autor(es): dc.contributorUniversidade Estadual de Campinas (UNICAMP)-
Autor(es): dc.creatorSuzuki, Miriam Fussae-
Autor(es): dc.creatorOliveira, João Ezequiel-
Autor(es): dc.creatorDamiani, Renata-
Autor(es): dc.creatorLima, Eliana Rosa-
Autor(es): dc.creatorAmaral, Kleicy Cavalcante-
Autor(es): dc.creatorSantos, Anderson Maikon de Souza [UNESP]-
Autor(es): dc.creatorMagalhães, Geraldo Santana-
Autor(es): dc.creatorFaverani, Leonardo Perez [UNESP]-
Autor(es): dc.creatorPereira, Luis Antonio Violin Dias-
Autor(es): dc.creatorSilva, Fabiana Medeiros-
Autor(es): dc.creatorBartolini, Paolo-
Data de aceite: dc.date.accessioned2022-02-22T00:24:52Z-
Data de disponibilização: dc.date.available2022-02-22T00:24:52Z-
Data de envio: dc.date.issued2020-12-11-
Data de envio: dc.date.issued2020-12-11-
Data de envio: dc.date.issued2020-11-30-
Fonte completa do material: dc.identifierhttp://dx.doi.org/10.1186/s13568-020-0964-5-
Fonte completa do material: dc.identifierhttp://hdl.handle.net/11449/198547-
Fonte: dc.identifier.urihttp://educapes.capes.gov.br/handle/11449/198547-
Descrição: dc.descriptionCommercially 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.-
Descrição: dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)-
Descrição: dc.descriptionBiotechnology Center Instituto de Pesquisas Energéticas e Nucleares IPEN-CNEN/SP, Avenida Prof. Lineu Prestes 2242, Cidade Universitária-
Descrição: dc.descriptionBiosintesis P&D-
Descrição: dc.descriptionDepartment of Surgery and Integrated Clinic-Universidade Estadual Paulista Júlio de Mesquita Filho-UNESP School of Dentistry-
Descrição: dc.descriptionImmunopathology Laboratory Instituto Butantan-
Descrição: dc.descriptionDepartment of Biochemistry and Tissue Biology Institute of Biology Universidade Estadual de Campinas-UNICAMP-
Descrição: dc.descriptionDepartment of Surgery and Integrated Clinic-Universidade Estadual Paulista Júlio de Mesquita Filho-UNESP School of Dentistry-
Descrição: dc.descriptionFAPESP: 2015/15446-0-
Descrição: dc.descriptionFAPESP: 2016/24724-6-
Idioma: dc.languageen-
Relação: dc.relationAMB Express-
???dc.source???: dc.sourceScopus-
Palavras-chave: dc.subjectBMP-2-
Palavras-chave: dc.subjectC2C12 bioassay-
Palavras-chave: dc.subjectCHO cell-derived-
Palavras-chave: dc.subjectEfficient osteoinductor-
Palavras-chave: dc.subjectEscherichia coli-derived-
Título: dc.titleHuman bone morphogenetic protein-2 (hBMP-2) characterization by physical–chemical, immunological and biological assays-
Tipo de arquivo: dc.typelivro digital-
Aparece nas coleções:Repositório Institucional - Unesp

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