Human bone morphogenetic protein‑2 (hBMP‑2) characterization by physical–chemical, immunological and biological assays
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KLEICY CAVALCANTE AMARAL
MIRIAM FUSSAE SUZUKI
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Commercially 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, sizeexclusion
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 methBMP-
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.
Como referenciar
SUZUKI, MIRIAM F.; OLIVEIRA, JOAO E.; DAMIANI, RENATA; LIMA, ELIANA R.; AMARAL, KLEICY C.; SANTOS, ANDERSON M. de S.; MAGALHÃES, GERALDO S.; FAVERANI, LEONARDO P.; PEREIRA, LUIS A.V.D.; SILVA, FABIANA M.; BARTOLINI, PAOLO. Human bone morphogenetic protein‑2 (hBMP‑2) characterization by physical–chemical, immunological and biological assays. AMB Express, v. 10, n. 1, p. 1-10, 2020. DOI: 10.1186/s13568-020-0964-5. Disponível em: http://repositorio.ipen.br/handle/123456789/31104. Acesso em: 24 Mar 2026.
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