Inductive properties of recombinant human BMP-2 produced in a bacterial expression system.
Recombinant human BMP-2, produced in E. coli, refolded and concentrated to a purity of more than 98%, has been demonstrated to be biologically active. In vitro, amounts of 0.4 microg BMP-2 or more induced new cartilage formation in 27 out of 47 samples of a neonatal muscle tissue assay, with chondroneogenesis occurring 14 days after a four-hour contact between BMP-2 and the muscle tissue. In vivo, BMP-2 was implanted in the thigh muscle of ICR mice for a period of three weeks. Amounts of 4 microg BMP-2 and more showed heterotopic bone formation in 15 out of 17 samples. When BMP-2 was combined with a collagen carrier, amounts of 0.4 microg protein or more induced heterotopic bone formation in 30 out of 33 samples four weeks after the implantation in the abdominal wall of Sprague-Dawley rats. The results show that the E. coli-derived BMP-2 was active in different assay systems in concentrations equal to those required with mammalian cell-expressed BMP-2. It could also be demonstrated that a single morphogen (BMP-2) is enough to initiate the differentiation process associated with bone induction. The presented bacterial expression system also offers the opportunity to produce large quantities of recombinant BMP-2 for clinical applications.
Kübler
N R
NR
Department of Cranio-Maxillo-Facial Surgery, School of Medicine, University of Würzburg, Germany.
Reuther
J F
JF
Faller
G
G
Kirchner
T
T
Ruppert
R
R
Sebald
W
W
D000009:Abdominal Muscles / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000008:administration & dosage / Q000235:genetics / Q000494:pharmacology*; D002356:Cartilage / Q000187:drug effects / Q000378:metabolism; D002454:Cell Differentiation; D019902:Chondrocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D003094:Collagen; D046508:Culture Techniques; D004305:Dose-Response Relationship, Drug; D004337:Drug Carriers; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D008648:Mesoderm / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008813:Mice, Inbred ICR; D018482:Muscle, Skeletal / Q000187:drug effects / Q000378:metabolism; D009999:Ossification, Heterotopic / Q000209:etiology / Q000378:metabolism / Q000473:pathology; D010012:Osteogenesis / Q000187:drug effects; D011506:Proteins / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta / Q000008:administration & dosage / Q000235:genetics / Q000494:pharmacology*
C525718:BMP2 protein, human; C525719:Bmp2 protein, mouse; C525720:Bmp2 protein, rat; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D004337:Drug Carriers; D011506:Proteins; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; C499469:recombinant human bone morphogenetic protein-2; D003094:Collagen