Synthesis of human bone morphogenetic protein-2 (hBMP-2) in E. coli periplasmic space
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Human BMP-2, a homodimeric protein that belongs to the TGF- 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.
Como referenciar
OLIVEIRA, JOAO E.; SUZUKI, MIRIAM F.; DAMIANI, RENATA; LIMA, ELIANA R.; AMARAL, KLEICY C.; SANTOS, ANDERSON M.S.; MAGALHAES, GERALDO S.; FAVERANI, LEONARDO P.; PEREIRA, LUIS A.V.D.; BARTOLINI, PAOLO. Synthesis of human bone morphogenetic protein-2 (hBMP-2) in E. coli periplasmic space: its characterization and preclinical testing. Cells, v. 10, n. 12, p. 1-16, 2021. DOI: 10.3390/cells10123525. Disponível em: http://repositorio.ipen.br/handle/123456789/32786. Acesso em: 26 Mar 2026.
Esta referência é gerada automaticamente de acordo com as normas do estilo IPEN/SP (ABNT NBR 6023) e recomenda-se uma verificação final e ajustes caso necessário.