title: Expression, purification, and renaturation of bone morphogenetic protein-2 from Escherichia coli.
abstract: Homodimeric bone morphogenetic protein-2 (BMP-2) is a member of the transforming growth factor beta superfamily that has been used for bone grafting. We were interested in exploring the functions of BMP-2 in other disease areas and focused on expressing and purifying active BMP-2 proteins. We have developed a new approach which involves using FoldIt refolding buffer to refold BMP-2 followed by a heparin affinity column to separate correctly folded dimer from monomer. A high yield of 29.4 mg BMP-2 dimer per gram cell wet weight was achieved. The purified BMP-2 dimer was shown to possess the same level of activity as BMP-2 from CHO cells as tested by the induction of alkaline phosphatase activity in C2C12 cells. This approach has potential application in refolding and purifying other homodimeric proteins.
authors.affiliation: Immusol, Inc., 10790 Roselle Street, San diego, CA 92121, USA. long@immusol.com
authors.forename: Shinong
authors.initials: S
authors.lastname: Long
authors.forename: Lynn
authors.initials: L
authors.lastname: Truong
authors.forename: Krista
authors.initials: K
authors.lastname: Bennett
authors.forename: Andrew
authors.initials: A
authors.lastname: Phillips
authors.forename: Flossie
authors.initials: F
authors.lastname: Wong-Staal
authors.forename: Hongwen
authors.initials: H
authors.lastname: Ma
mesh_terms: D000818:Animals; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification*; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004926:Escherichia coli; D015870:Gene Expression; D006801:Humans; D002479:Inclusion Bodies / Q000737:chemistry; D017510:Protein Folding; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification*; D016212:Transforming Growth Factor beta / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta