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Genzel Y.,Max Planck Institute for Dynamics of Complex Technical Systems | Vogel T.,Max Planck Institute for Dynamics of Complex Technical Systems | Vogel T.,Esslingen University of Applied Sciences | Buck J.,Max Planck Institute for Dynamics of Complex Technical Systems | And 7 more authors.
Vaccine | Year: 2014

High cell densities in animal cell culture can be obtained by continuous perfusion of fresh culture medium across hollow fiber membranes that retain the cells. Careful selection of the membrane type and cut-off allows to control accumulation of target molecules and removal of low molecular weight compounds. In this report, perfusion with the scalable ATF (alternating tangential filtration, Refine Technology) system was evaluated for two suspension cell lines, the avian cell line AGE1.CR and the human cell line CAP. Both were cultivated in chemically defined media optimized for batch cell growth in a 1. L stirred tank bioreactor connected to the smallest ATF unit (ATF2) and infected with cell line-adapted human influenza A virus (A/PR/8/34 (H1N1), typical diameter: 80-100. nm).At concentrations of about 25 million cells/mL three different membrane cut-offs (50. kDa, 0.2. μm and 0.5. μm) were tested and compared to batch cultivations performed at 5 million cells/mL. For medium and large cut-offs no cell-density effect could be observed with cell-specific virus yields of 1428-1708 virions/AGE1.CR cell (infected with moi 0.001) and 1883-4086 virions/CAP cell (moi of 0.025) compared to 1292 virions/AGE1.CR cell and 3883 virions/CAP cell in batch cultures. Even at a concentration of 48 million AGE1.CR cells/mL (cut-off: 0.2. μm) a cell-specific yield of 1266 virions/cell was reached. Only for the small cut-off (50. kDa) used with AGE1.CR cells a decrease in cell-specific yield was measured with 518 virions/cell. Surprisingly, the ratio of infectious to total virions seemed to be increased in ATF compared to batch cultures. AGE1.CR cell-derived virus particles were present in the permeate (0.2 and 0.5. μm cut-off), whereas CAP cell-derived virions were not, suggesting possible differences in morphology, aggregation or membrane properties of the virions released by the two cell lines.To our knowledge, this is the first study that illustrates the potential of ATF-based perfusion of chemically defined media across cell-retaining membranes for production of an influenza A vaccine. © 2014 Elsevier Ltd. Source


Genzel Y.,Max Planck Institute for Dynamics of Complex Technical Systems | Behrendt I.,Max Planck Institute for Dynamics of Complex Technical Systems | Rodig J.,Max Planck Institute for Dynamics of Complex Technical Systems | Rapp E.,Max Planck Institute for Dynamics of Complex Technical Systems | And 5 more authors.
Applied Microbiology and Biotechnology | Year: 2013

Forced by major drawbacks of egg-based influenza virus production, several studies focused on the establishment and optimization of cell-based production systems. Among numerous possible host cell lines from duck, monkey, canine, chicken, mouse, and human origin, only a few will meet regulatory requirements, accomplish industrial standards, and result in high virus titers. From primary virus isolation up to large-scale manufacturing of human vaccines, however, the most logical choice seems to be the use of human cell lines. For this reason, we evaluated the recently established CAP cell line derived from human amniocytes for its potential in influenza virus production in suspension culture in small scale shaker flask and stirred tank bioreactor experiments. Different human and animal influenza viruses could be adapted to produce hemagglutination (HA) titers of at least 2.0 log10 HA units/100 μL without further process optimization. Adjusting trypsin activity as well as infection conditions (multiplicity of infection, infection medium) resulted in HA titers of up to 3.2 log10 HA units/100 μL and maximum cell-specific virus productivities of 6,400 virions/cell (for human influenza A/PR/8/34 as a reference). Surface membrane expression of sialyloligosaccharides as well as HA N-glycosylation patterns were characterized. Overall, experimental results clearly demonstrate the potential of CAP cells for achieving high virus yields for different influenza strains and the option to introduce a highly attractive fully characterized human cell line compliant with regulatory and industrial requirements as an alternative for influenza virus vaccine production. © 2012 Springer-Verlag. Source


Trademark
CEVEC Pharmaceuticals GmbH | Date: 2008-04-01

Chemical preparations for scientific purposes and biological products for scientific purposes in the nature of microorganisms, eukaryotic cell culture lines, polypeptides, proteins, nucleic acids, viral vectors and gene expression vectors, except for medical and veterinary purposes; chemical and biological preparations other than for medical or veterinary purposes, for use in research and characterization of diseases, namely, diagnostic preparations for scientific and research use; cultures of microorganisms for scientific purposes, eukaryotic cell culture lines for scientific purposes. Medicaments for the treatment of cancer, tumorous diseases, proliferative diseases, allergies, auto-immune diseases, inflammation, inflammatory diseases, infectious diseases, osteoporosis, graft versus host and host versus graft diseases, transplant rejection, coronary heart diseases, minimal residual diseases, restenosis, metabolic disorders, neurological disorders, diabetes, B cell malignancy, rheumatoid arthritis, immunological disorders, parasitic reactions, skin diseases, neurodegenerative diseases and cardio-vascular diseases; pharmaceutical and veterinary preparations for use in the treatment of cancer, tumorous diseases, proliferative diseases, allergies, auto-immune diseases, inflammation, inflammatory diseases, infectious diseases, osteoporosis, graft versus host and host versus graft diseases, transplant rejection, coronary heart diseases, minimal residual diseases, restenosis, metabolic disorders, neurological disorders, diabetes, B cell malignancy, rheumatoid arthritis, immunological disorders, parasitic reactions, skin diseases, neurodegenerative diseases and cardio-vascular diseases; chemical and biological preparations for medical use, namely, for medical diagnosis and analysis; eukaryotic cell culture lines, microorganisms, polypeptides, nucleic acids for medical and medical diagnostic purposes. Scientific and industrial research and consulting services, in particular in the fields of molecular biology, biotechnology, genetics; pharmaceutical research and development; medical research and development of methods of medical therapy; scientific and technological services in the nature of breeding, recovery, optimization, synthesis and modification of eukaryotic cell culture lines, micro-organisms, polypeptides, nucleic acids for third parties; research services in the fields of medicine and pharmaceuticals; medical and scientific research, namely, conducting clinical trials for third parties. Medical and veterinary services; medical services, namely, breeding, recovery, optimization, synthesis and modification of eukaryotic cell culture lines, micro-organisms, polypeptides, nucleic acids for third parties; consulting services in the fields of medicine and pharmaceuticals.


Patent
CEVEC Pharmaceuticals GmbH | Date: 2014-12-19

The present invention relates to a method for the production of a permanent human cell line, wherein isolated primary human cells are transfected simultaneously with a sequence allowing the expression of at least one cell transforming factor and a sequence allowing the expression of at least one recombinant polypeptide.


Patent
CEVEC Pharmaceuticals GmbH | Date: 2011-08-16

The present invention relates to a method for the production of an influenza virus-based vaccine using permanent human amniocyte cells, as well as the use of a permanent human amniocyte cell for the production of a influenza virus-based vaccine.

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