Synthesis of human bone morphogenetic protein-2 (Hbmp-2) in e. coli periplasmic space: its characterization and preclinical testing

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MetadadosDescriçãoIdioma
Autor(es): dc.contributorIPEN–CNEN-
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.creatorOliveira, João E.-
Autor(es): dc.creatorSuzuki, Miriam F.-
Autor(es): dc.creatorDamiani, Renata-
Autor(es): dc.creatorLima, Eliana R.-
Autor(es): dc.creatorAmaral, Kleicy C.-
Autor(es): dc.creatorSantos, Anderson M. S.-
Autor(es): dc.creatorMagalhães, Geraldo S.-
Autor(es): dc.creatorFaverani, Leonardo P.-
Autor(es): dc.creatorPereira, Luís A. V. D.-
Autor(es): dc.creatorBartolini, Paolo-
Data de aceite: dc.date.accessioned2025-08-21T17:46:18Z-
Data de disponibilização: dc.date.available2025-08-21T17:46:18Z-
Data de envio: dc.date.issued2022-04-28-
Data de envio: dc.date.issued2022-04-28-
Data de envio: dc.date.issued2021-11-30-
Fonte completa do material: dc.identifierhttp://dx.doi.org/10.3390/cells10123525-
Fonte completa do material: dc.identifierhttp://hdl.handle.net/11449/223080-
Fonte: dc.identifier.urihttp://educapes.capes.gov.br/handle/11449/223080-
Descrição: dc.descriptionHuman BMP-2, a homodimeric protein that belongs to the TGF-β family, is a recognized osteoinductor due to its capacity of inducing bone regeneration and ectopic bone formation. The administration of its recombinant form is an alternative to autologous bone grafting. A variety of E. coli-derived hBMP-2 has been synthesized through refolding of cytoplasmic inclusion bodies. The present work reports the synthesis, purification, and characterization of periplasmic hBMP-2, obtained directly in its correctly folded and authentic form, i.e., without the initial methionine typical of the cytoplasmic product that can induce undesired immunoreactivity. A bacterial expression vector was constructed including the DsbA signal peptide and the cDNA of hBMP-2. The periplasmic fluid was extracted by osmotic shock and analyzed via SDS-PAGE, Western blotting, and reversed-phase high-performance liquid chromatography (RP-HPLC). The purification was carried out by heparin affinity chromatography, followed by high-performance size-exclusion chromatography (HPSEC). HPSEC was used for qualitative and quantitative analysis of the final product, which showed >95% purity. The classical in vitro bioassay based on the induction of alkaline phosphatase activity in myoblastic murine C2C12 cells and the in vivo bioassay consisting of treating calvarial critical-size defects in rats confirmed its bioactivity, which matched the analogous literature data for hBMP-2.-
Descrição: dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)-
Descrição: dc.descriptionInstituto de Pesquisas Energéticas e Nucleares IPEN–CNEN, Av. Prof. Lineu Prestes 2242-
Descrição: dc.descriptionBiosintesis P & D-
Descrição: dc.descriptionDepartment of Surgery and Integrated Clinic School of Dentistry Universidade Estadual Paulista Júlio de Mesquita Filho UNESP-
Descrição: dc.descriptionImmunopathology Laboratory Instituto Butantan-
Descrição: dc.descriptionDepartment of Biochemistry and Tissue Biology Institute of Biology State University of Campinas UNICAMP-
Descrição: dc.descriptionDepartment of Surgery and Integrated Clinic School of Dentistry Universidade Estadual Paulista Júlio de Mesquita Filho UNESP-
Descrição: dc.descriptionFAPESP: 2015/15446-0-
Descrição: dc.descriptionFAPESP: 2016/24724-6-
Idioma: dc.languageen-
Relação: dc.relationCells-
???dc.source???: dc.sourceScopus-
Palavras-chave: dc.subjectCalvarial critical-size defect-
Palavras-chave: dc.subjectHBMP-2-
Palavras-chave: dc.subjectOsteoinductor-
Palavras-chave: dc.subjectPeriplasmic expression-
Título: dc.titleSynthesis of human bone morphogenetic protein-2 (Hbmp-2) in e. coli periplasmic space: its characterization and preclinical testing-
Tipo de arquivo: dc.typelivro digital-
Aparece nas coleções:Repositório Institucional - Unesp

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