Renaturation and purification of bone morphogenetic protein-2 produced as inclusion bodies in high-cell-density cultures of recombinant Escherichia coli

Transforming Growth Factor-β-activated Kinase-1 (TAK1), a MAP3K, Interacts with Smad Proteins and Interferes with Osteogenesis in Murine Mesenchymal Progenitors

“…Recombinant human BMP4 and TGF-␤1 were purchased from R&D Systems (Wiesbaden, Germany), and recombinant human IL-1␤ was from RELIATech GmbH (Braunschweig, Germany). Recombinant human BMP2 from Escherichia coli was prepared as described (30). C3H10T 1 ⁄2-BMP2 cells were stably transfected with expression plasmids encoding wt TAK1, dominant-negative (dn) or constitutively active (ca) TAK1 in pMT7T3-f1 using DOSPER TM according to the manufacturer's protocol (Roche Applied Science) together with a selection plasmid conferring puromycin resistance (pBSpac⌬p, parental C3H10T 1 ⁄2 cells) or G418 resistance (pAG60, BMP2-expressing C3H10T 1 ⁄2 cells).…”

Soluble expression and purification of synthetic human bone morphogenetic protein-2 in Escherichia coli

“…Recombinant BMPs have been obtained from mammalian cell cultures, but incomplete monomer processing and low yields are usual in these processes (6). Alternatively, the production of biologically active recombinant humans BMPs (rhBMPs) through in vitro refolding of Escherichia coli produced inclusion bodies has been reported (4,7). However, the refolding procedure is complicated and the overall yield is low.…”

Different cell populations are inducible by BMP-2 covalently covered Bioverit® II implants in rabbit subcutis and middle ear

“…After drying at 60°C for 30 min the implants were dipped again until the coverage consisted of three layers of silica. After calcination at 415°C for 4 h the samples were incubated with 10 % 3-aminopropyl trimethoxysilane (Sigma-Aldrich Chemie) in water for 2 min, dried and shaking in a solution of 250 µg ml -1 isolated recombinant human BMP-2 (see method in [5]) in 50 mM 2(Nmorpholino)ethanesulfonic acid buffer, pH 5 (SigmaAldrich Chemie) overnight at 4°C under gentle shaking, washed with 0.066% sodium dodecyl sulphate in 0.125 M sodium tetraborate buffer, pH10.0 (Sigma-Aldrich Chemie), transferred in phosphate-buffered saline (Gibco tm, Invitrogen Corporation, Paisley, UK) and implanted immediately.…”