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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

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Suzuki, Miriam Fussae [1] ; Oliveira, Joao Ezequiel [1] ; Damiani, Renata [2] ; Lima, Eliana Rosa [1] ; Amaral, Kleicy Cavalcante [1] ; Santos, Anderson Maikon de Souza [3, 4] ; Magalhaes, Geraldo Santana [5] ; Faverani, Leonardo Perez [3, 4] ; Pereira, Luis Antonio Violin Dias [6] ; Silva, Fabiana Medeiros [2] ; Bartolini, Paolo [1]
Total Authors: 11
Affiliation:
[1] IPEN CNEN SP, Inst Pesquisas Energet & Nucl, Ctr Biotechnol, Ave Prof Lineu Prestes 2242, BR-05508000 Sao Paulo, SP - Brazil
[2] Biosintesis P&D, Sao Paulo, SP - Brazil
[3] Univ Estadual Paulista Julio de Mesquita Filho UN, Sch Dent, Dept Surg, Aracatuba, SP - Brazil
[4] Univ Estadual Paulista Julio de Mesquita Filho UN, Sch Dent, Integrated Clin, Aracatuba, SP - Brazil
[5] Inst Butantan, Immunopathol Lab, Sao Paulo, SP - Brazil
[6] Univ Estadual Campinas UNICAMP, Inst Biol, Dept Biochem & Tissue Biol, Campinas, SP - Brazil
Total Affiliations: 6
Document type: Journal article
Source: AMB EXPRESS; v. 10, n. 1 FEB 17 2020.
Web of Science Citations: 0
Abstract

Commercially available preparations of methionyl-human BMP-2 and CHO-derived hBMP-2, which belongs to the transforming growth factor beta (TGF-beta) 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. (AU)

FAPESP's process: 16/24724-6 - Expression, purification and characterization of bone morphogenetic protein (BMP-2) for application in composites biomaterials with poly(E-caprolactone)
Grantee:Paolo Bartolini
Support type: Research Grants - Innovative Research in Small Business - PIPE
FAPESP's process: 15/15446-0 - Expression, purification and characterization of bone morphogenetic protein (BMP-2) for application in composites biomaterials with poly(E-caprolactone)
Grantee:Paolo Bartolini
Support type: Research Grants - Innovative Research in Small Business - PIPE