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Continuous separation of the monomeric and dimeric forms of human bone morphogenetic protein-2 (BMP-2) using gradient Simulated Moving Bed (SMB) chromatography

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http://pubman.mpdl.mpg.de/cone/persons/resource/persons86311

Gueorguieva,  L.
Physical and Chemical Foundations of Process Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Max Planck Society;
Otto-von-Guericke-Universität Magdeburg, External Organizations;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons86477

Seidel-Morgenstern,  A.
Physical and Chemical Foundations of Process Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Max Planck Society;
Otto-von-Guericke-Universität Magdeburg, External Organizations;

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Citation

Gueorguieva, L., Seidel-Morgenstern, A., Vallejo, L. F., & Rinas, U. (2004). Continuous separation of the monomeric and dimeric forms of human bone morphogenetic protein-2 (BMP-2) using gradient Simulated Moving Bed (SMB) chromatography. Poster presented at ISPPP 2004: International Symposium on the Separation of Proteins, Peptides and Polynucleotides, Aachen, Germany.


Cite as: http://hdl.handle.net/11858/00-001M-0000-0013-9D81-9
Abstract
Bone morphogenetic protein-2 (BMP-2) is one of the most interesting of the approx. 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. BMP-2 has been produced in form of inactive inclusion bodies in E. coli [1] and after solubilization and renaturation, the biologically active dimeric form of BMP-2 is generated [2]. However, inactive monomers of BMP-2 are also formed during the renaturation process which need to be separated from the active dimeric BMP-2. Goal of the project is the development of a continuous gradient SMB-process [3] exploiting two different salt concentrations in the feed solution and in the desorbent. A three-zone-configuration is designed based on a systematic study of the adsorption equilibrium of the monomers and dimers of BMP-2 at different salt concentrations. These equilibrium data allow to specify suitable salt concentrations and flow rates of the SMB-process offering the potential to collect the better adsorbed dimer with high purity at the extract port of the unit. In this presentation are presented: 1) A theoretical study of the three-zone-gradient-SMB process 2) The current state of the experimental work [1] Vallejo, L.F., Brokelmann, M., Marten, S., Trappe, S., Cabrera-Crespo, J., Hoffmann, A., Gross, G., Weich, H.A. and Rinas, U. (2002) Renaturation and purification of bone morphogenetic protein-2 produced as inclusion bodies in high-cell-density cultures of recombinant Escherichia coli. J. Biotechnol., 94, 185-194 [2] Vallejo, L.F. and Rinas, U. (2004) Optimized procedure for renaturation of recombinant human bone morphogenetic protein-2 at high protein concentration. Biotechnol. Bioeng., 85, 601-609 [3] Antos, D., Seidel-Morgenstern, A. (2001) Application of gradients in the simulated moving bed processes. Chem. Eng. Sci. 56, 6667-6682