Details
| Original language | English |
|---|---|
| Pages (from-to) | 142-150 |
| Number of pages | 9 |
| Journal | Journal of Chromatography A |
| Volume | 1135 |
| Issue number | 2 |
| Early online date | 24 Oct 2006 |
| Publication status | Published - 1 Dec 2006 |
| Externally published | Yes |
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-β (TGF-β) 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.
Keywords
- Bone morphogenetic protein-2, Continuous counter-current separation, Gradient elution, Step gradients
ASJC Scopus subject areas
- Chemistry(all)
- Analytical Chemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Chemistry(all)
- Organic Chemistry
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In: Journal of Chromatography A, Vol. 1135, No. 2, 01.12.2006, p. 142-150.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Discontinuous and continuous separation of the monomeric and dimeric forms of human bone morphogenetic protein-2 from renaturation batches
AU - Gueorguieva, Ludmila
AU - Vallejo, Luis Felipe
AU - Rinas, Ursula
AU - Seidel-Morgenstern, Andreas
N1 - Funding Information: The financial support of Deutsche Forschungsgemeinschaft/Sonderforschungsbereich (SFB) 578 and Fonds der Chemischen Industrie is gratefully acknowledged.
PY - 2006/12/1
Y1 - 2006/12/1
N2 - Bone morphogenetic protein-2 (BMP-2) is one of the most interesting of the approximately 14 BMPs which belong to the transforming-growth-factor-β (TGF-β) 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.
AB - Bone morphogenetic protein-2 (BMP-2) is one of the most interesting of the approximately 14 BMPs which belong to the transforming-growth-factor-β (TGF-β) 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.
KW - Bone morphogenetic protein-2
KW - Continuous counter-current separation
KW - Gradient elution
KW - Step gradients
UR - http://www.scopus.com/inward/record.url?scp=33750843738&partnerID=8YFLogxK
U2 - 10.1016/j.chroma.2006.08.061
DO - 10.1016/j.chroma.2006.08.061
M3 - Article
C2 - 17064713
AN - SCOPUS:33750843738
VL - 1135
SP - 142
EP - 150
JO - Journal of Chromatography A
JF - Journal of Chromatography A
SN - 0021-9673
IS - 2
ER -