Biotinated bone morphogenetic protein-2: In vivo and in vitro activity.
Recombinant human bone morphogenetic protein-2 (rhBMP-2) was biotinated, and the bioactivity of biotinated protein was assessed in vitro (alkaline phosphate induction in limb bud cells) and in vivo (osteoinduction in the rat ectopic assay). Amino-biotinated rhBMP-2 exhibited an increase in bioactivity whereas carboxy-biotinated rhBMP-2 did not exhibit any changes in bioactivity in vitro. Avidin inhibited the bioactivity of amino-biotinated but not carboxyl-biotinated rhBMP-2. Both amino- and carboxy-modified rhBMP-2 induced bone at an equivalent level to that of unmodified rhBMP-2 in vivo. The presence of avidin did not affect the osteoinductive activity of both types of biotinated rhBMP-2. The overall results indicated that binding to a large protein, avidin, might affect rhBMP-2 activity in vitro depending on the binding site; however, in vivo activity was unaffected by the avidin binding.
Uludag
H
H
Genetics Institute, Bone Biology and Application, One Burtt Road, Andover, Massachusetts 01810, USA. hasan.uludag@yakberta.ca
Golden
J
J
Palmer
R
R
Wozney
J M
JM
D000818:Animals; D001360:Avidin / Q000378:metabolism / Q000494:pharmacology; D001665:Binding Sites; D019921:Biotinylation / Q000379:methods; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism* / Q000493:pharmacokinetics / Q000494:pharmacology*; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D004343:Drug Implants; D006801:Humans; D018878:Limb Buds / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008970:Molecular Weight; D010012:Osteogenesis / Q000187:drug effects*; D051381:Rats; D020318:Rats, Long-Evans; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D016212:Transforming Growth Factor beta
C525718:BMP2 protein, human; C525720:Bmp2 protein, rat; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D004343:Drug Implants; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; C499469:recombinant human bone morphogenetic protein-2; D001360:Avidin