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

Publikation: Beitrag in Fachzeitschrift › Artikel › Forschung › Peer-Review

Autorschaft

  • Ludmila Gueorguieva
  • Luis Felipe Vallejo
  • Ursula Rinas
  • Andreas Seidel-Morgenstern

Externe Organisationen

  • Otto-von-Guericke-Universität Magdeburg
  • Helmholtz-Zentrum für Infektionsforschung GmbH (HZI)
  • Max-Planck-Institut für Dynamik komplexer technischer Systeme
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Details

OriginalspracheEnglisch
Seiten (von - bis)142-150
Seitenumfang9
FachzeitschriftJournal of Chromatography A
Jahrgang1135
Ausgabenummer2
Frühes Online-Datum24 Okt. 2006
PublikationsstatusVeröffentlicht - 1 Dez. 2006
Extern publiziertJa

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.

ASJC Scopus Sachgebiete

Zitieren

Discontinuous and continuous separation of the monomeric and dimeric forms of human bone morphogenetic protein-2 from renaturation batches. / Gueorguieva, Ludmila; Vallejo, Luis Felipe; Rinas, Ursula et al.
in: Journal of Chromatography A, Jahrgang 1135, Nr. 2, 01.12.2006, S. 142-150.

Publikation: Beitrag in Fachzeitschrift › Artikel › Forschung › Peer-Review

Gueorguieva L, Vallejo LF, Rinas U, Seidel-Morgenstern A. Discontinuous and continuous separation of the monomeric and dimeric forms of human bone morphogenetic protein-2 from renaturation batches. Journal of Chromatography A. 2006 Dez 1;1135(2):142-150. Epub 2006 Okt 24. doi: 10.1016/j.chroma.2006.08.061
Gueorguieva, Ludmila ; Vallejo, Luis Felipe ; Rinas, Ursula et al. / Discontinuous and continuous separation of the monomeric and dimeric forms of human bone morphogenetic protein-2 from renaturation batches. in: Journal of Chromatography A. 2006 ; Jahrgang 1135, Nr. 2. S. 142-150.
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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.",
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AU - Seidel-Morgenstern, Andreas

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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.

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