tetano
Editor, Senior Moderator
J Chromatogr A. 2013 Jul 26. pii: S0021-9673(13)01160-6. doi: 10.1016/j.chroma.2013.07.081. [Epub ahead of print]
Continuous purification of influenza virus using simulated moving bed chromatography.
Kr?ber T, Wolff MW, Hundt B, Seidel-Morgenstern A, Reichl U.
Source
Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstra?e 1, 39106 Magdeburg, Germany. Electronic address: kroeber@mpi-magdeburg.mpg.de.
Abstract
Continuous size exclusion chromatography for the separation of cell culture-derived influenza virus from contaminating proteins was established successfully. Therefore, an open loop simulated moving bed (SMB) setup with one column per zone was applied. Several operating conditions were tested and overall trends were found to be in agreement with expectations derived from theory. Furthermore, the separation performance was compared to an optimized conventional batch chromatography. The yield of influenza virus in the product fraction, based on a hemagglutination assay, was 70% (SMB) and 80% (batch), respectively. The amount of contaminating protein per product was 0.61μgkHAU-1 (SMB) compared to 0.29μgkHAU-1 (batch). This corresponds to a reduction of the respective amount in the feed solution by 60% and 80%, respectively. For both processes, the estimated amount of total protein per vaccine dose would meet the level required for manufacturing of human influenza vaccines prepared in cell cultures. Depending on the strategy chosen for sanitization and equilibration of columns the calculated overall productivity for the SMB process was up to 3.8 times higher compared to the batch mode. SMB, therefore, has the potential to replace single column discontinuous chromatography in order to design more efficient purification trains for production of cell culture-derived influenza vaccines.
Copyright ? 2013 Elsevier B.V. All rights reserved.
KEYWORDS:
Downstream processing, Influenza virus, Simulated moving bed, Size exclusion chromatography, Vaccine purification
PMID:
23932371
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/23932371
Continuous purification of influenza virus using simulated moving bed chromatography.
Kr?ber T, Wolff MW, Hundt B, Seidel-Morgenstern A, Reichl U.
Source
Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstra?e 1, 39106 Magdeburg, Germany. Electronic address: kroeber@mpi-magdeburg.mpg.de.
Abstract
Continuous size exclusion chromatography for the separation of cell culture-derived influenza virus from contaminating proteins was established successfully. Therefore, an open loop simulated moving bed (SMB) setup with one column per zone was applied. Several operating conditions were tested and overall trends were found to be in agreement with expectations derived from theory. Furthermore, the separation performance was compared to an optimized conventional batch chromatography. The yield of influenza virus in the product fraction, based on a hemagglutination assay, was 70% (SMB) and 80% (batch), respectively. The amount of contaminating protein per product was 0.61μgkHAU-1 (SMB) compared to 0.29μgkHAU-1 (batch). This corresponds to a reduction of the respective amount in the feed solution by 60% and 80%, respectively. For both processes, the estimated amount of total protein per vaccine dose would meet the level required for manufacturing of human influenza vaccines prepared in cell cultures. Depending on the strategy chosen for sanitization and equilibration of columns the calculated overall productivity for the SMB process was up to 3.8 times higher compared to the batch mode. SMB, therefore, has the potential to replace single column discontinuous chromatography in order to design more efficient purification trains for production of cell culture-derived influenza vaccines.
Copyright ? 2013 Elsevier B.V. All rights reserved.
KEYWORDS:
Downstream processing, Influenza virus, Simulated moving bed, Size exclusion chromatography, Vaccine purification
PMID:
23932371
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/23932371