Expression,Purification, Bioactivity,and Partial Characterization of a Recombinant Human Bone Morphogenetic Protein-7 Produced in Human 293T Cells |
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| Authors: | J. C. Bustos-Valenzuela E. Halcsik Ê. J. Bassi M. A. Demasi J. M. Granjeiro M. C. Sogayar |
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| Affiliation: | 1.NUCEL – Cell and Molecular Therapy Center,University of S?o Paulo,S?o Paulo,Brazil;2.Institute of Chemistry,University of S?o Paulo,S?o Paulo,Brazil;3.Cell and Molecular Biology Department,Federal Fluminense University,Niterói,Brazil |
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| Abstract: | Bone morphogenetic protein-7 (BMP-7) is a secreted multifunctional growth factor of the TGF-β superfamily, which is predominantly
known for its osteoinductive properties and emerging potential for treatment of kidney diseases. The mature 34–38 kDa disulfide-linked
homodimer protein plays a key role in the differentiation of mesenchymal cells into bone and cartilage. In this study, the
full-length sequence of hBMP-7 was amplified and, then, cloned, expressed, and purified from the conditioned medium of 293T
cells stably transfected with a lentiviral vector. The mature protein dimer form was properly secreted and recognized by anti-BMP-7
antibodies, and the protein was shown to be glycosilated by treatment with exoglycosidase, followed by western blotting. Moreover,
the activity of the purified protein was demonstrated both in vitro, by alkaline phosphatase activity in C2C12 cells, and
in vivo by induction of ectopic bone formation in Balb/c Nude mice after 21 days, respectively. This recombinant protein platform
may be very useful for expression of different human cytokines and other proteins for medical applications. |
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