Discontinuous and continuous separation of the monomeric and dimeric forms of human bone morphogenetic protein-2 from renaturation batches.

2.50
Hdl Handle:
http://hdl.handle.net/10033/15836
Title:
Discontinuous and continuous separation of the monomeric and dimeric forms of human bone morphogenetic protein-2 from renaturation batches.
Authors:
Gueorguieva, Ludmila; Vallejo, Luis Felipe; Rinas, Ursula; Seidel-Morgenstern, Andreas
Abstract:
Bone morphogenetic protein-2 (BMP-2) is one of the most interesting of the approximately 14 BMPs which belong to the transforming-growth-factor-beta (TGF-beta) superfamily. BMP-2 induces bone formation and thus plays an important role as a pharmaceutical protein. Recently, rhBMP-2 has been produced in form of inactive inclusion bodies in Escherichia coli. After solubilization and renaturation the biologically active dimeric form of rhBMP-2 can be generated. However, inactive monomers of BMP-2 are also formed during the renaturation process which must be separated from the active dimeric BMP-2. The purpose of this paper is to present: (a) results of an experimental study of a chromatographic separation of the monomeric and dimeric forms; and (b) a concept for a continuous counter-current simulated moving bed (SMB) process. The capacity of heparin as stationary phase was estimated for different salt concentrations in the mobile phase. A simulation study of a three-zone SMB process was performed applying a two step salt gradient. The results reveal the potential of the process for the purification of the dimeric BMP-2.
Affiliation:
Otto-von-Guericke-Universität Magdeburg, Institut für Verfahrenstechnik, PO Box 4120, D-39106 Magdeburg, Germany.
Citation:
Discontinuous and continuous separation of the monomeric and dimeric forms of human bone morphogenetic protein-2 from renaturation batches. 2006, 1135 (2):142-50notJ Chromatogr A
Journal:
Journal of chromatography. A
Issue Date:
1-Dec-2006
URI:
http://hdl.handle.net/10033/15836
DOI:
10.1016/j.chroma.2006.08.061
PubMed ID:
17064713
Type:
Article
Language:
en
ISSN:
0021-9673
Appears in Collections:
Publications from Division of Molekulare Struktur Biologie (MOSB)

Full metadata record

DC FieldValue Language
dc.contributor.authorGueorguieva, Ludmila-
dc.contributor.authorVallejo, Luis Felipe-
dc.contributor.authorRinas, Ursula-
dc.contributor.authorSeidel-Morgenstern, Andreas-
dc.date.accessioned2008-01-08T15:15:34Z-
dc.date.available2008-01-08T15:15:34Z-
dc.date.issued2006-12-01-
dc.identifier.citationDiscontinuous and continuous separation of the monomeric and dimeric forms of human bone morphogenetic protein-2 from renaturation batches. 2006, 1135 (2):142-50notJ Chromatogr Aen
dc.identifier.issn0021-9673-
dc.identifier.pmid17064713-
dc.identifier.doi10.1016/j.chroma.2006.08.061-
dc.identifier.urihttp://hdl.handle.net/10033/15836-
dc.description.abstractBone morphogenetic protein-2 (BMP-2) is one of the most interesting of the approximately 14 BMPs which belong to the transforming-growth-factor-beta (TGF-beta) superfamily. BMP-2 induces bone formation and thus plays an important role as a pharmaceutical protein. Recently, rhBMP-2 has been produced in form of inactive inclusion bodies in Escherichia coli. After solubilization and renaturation the biologically active dimeric form of rhBMP-2 can be generated. However, inactive monomers of BMP-2 are also formed during the renaturation process which must be separated from the active dimeric BMP-2. The purpose of this paper is to present: (a) results of an experimental study of a chromatographic separation of the monomeric and dimeric forms; and (b) a concept for a continuous counter-current simulated moving bed (SMB) process. The capacity of heparin as stationary phase was estimated for different salt concentrations in the mobile phase. A simulation study of a three-zone SMB process was performed applying a two step salt gradient. The results reveal the potential of the process for the purification of the dimeric BMP-2.en
dc.language.isoenen
dc.subject.meshBone Morphogenetic Proteinsen
dc.subject.meshChromatography, Liquiden
dc.subject.meshDimerizationen
dc.subject.meshElectrophoresis, Polyacrylamide Gelen
dc.subject.meshEscherichia colien
dc.subject.meshHeparinen
dc.subject.meshProtein Renaturationen
dc.subject.meshRecombinant Proteinsen
dc.subject.meshSpectrophotometry, Ultravioleten
dc.subject.meshTransforming Growth Factor betaen
dc.titleDiscontinuous and continuous separation of the monomeric and dimeric forms of human bone morphogenetic protein-2 from renaturation batches.en
dc.typeArticleen
dc.contributor.departmentOtto-von-Guericke-Universität Magdeburg, Institut für Verfahrenstechnik, PO Box 4120, D-39106 Magdeburg, Germany.en
dc.identifier.journalJournal of chromatography. Aen

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