SUZUKI, MIRIAM F.OLIVEIRA, JOAO E.DAMIANI, RENATALIMA, 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, PAOLO2020-04-072020-04-072020SUZUKI, 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. <b>AMB Express</b>, v. 10, n. 1, p. 1-10, 2020. DOI: <a href="https://dx.doi.org/10.1186/s13568-020-0964-5">10.1186/s13568-020-0964-5</a>. Disponível em: http://repositorio.ipen.br/handle/123456789/31104.2191-0855http://repositorio.ipen.br/handle/123456789/31104Commercially 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.1-10openAccessproteinsskeletonhuman populationsbone cellscho cellsbioassayescherichia coliconnective tissue cellsArtigo de periódico11010.1186/s13568-020-0964-50000-0002-6937-11200000-0002-7467-345753.7767.00