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  • 1
    Electronic Resource
    Electronic Resource
    An integrated strategy for the process development of a recombinant antibody-cytokine fusion protein expressed in BHK cells (1999)
    Springer
    Applied microbiology and biotechnology 52 (1999), S. 345-353 
    Details
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Recombinant fusion proteins offer important new therapeutic approaches for the future. This report describes the use of three different genetic strategies (i.e. “mono-”, “bi-” and “tri-cistronic” vectors) to achieve stable secretion from BHK cells of a glycosylated antibody-cytokine fusion protein designed for use in anti-tumour therapy. It describes selection of a robust and effective production cell line based on stability of secretion of the product, quality of mRNA and protein products and performance in in vitro bioassays for potency. The data obtained at this stage were utilised in the selection of a suitable candidate production cell line. The relative productivity and general performance of the cells in stirred tank and fixed bed culture systems indicated that a variety of cell culture technologies provided robust tools for production of a highly selected cell clone. Consistency of the product glycosylation was determined by analysis of released oligosaccharides using matrix-assisted laser desorption ionisation – time of flight mass spectrometry and high-performance anion exchange chromatography. These investigations showed consistent expression of three glycoforms of the fusion protein which varied in their relative proportions in different culture systems and at different time points in a fixed bed reactor with continuous perfusion. In conclusion, this study dealt with a range of important scientific and technical issues which are essential for regulatory approval and commercial success of a recombinant protein and elucidates some useful markers for process development for similar recombinant biologicals.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002530051530
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    Immobilization of animal cells in porous carrier culture
    Amsterdam : Elsevier
    Trends in Biotechnology 8 (1990), S. 204-209 
    Details
    ISSN: 0167-7799
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
    URL: http://linkinghub.elsevier.com/retrieve/pii/0167-7799(90)90177-Y
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    Paper (German National Licenses)
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  • 3
    Electronic Resource
    Electronic Resource
    Use of lactate dehydrogenase release to assess changes in culture viability (1990)
    Springer
    Cytotechnology 3 (1990), S. 301-307 
    Details
    ISSN: 1573-0778
    Keywords: culture viability ; lactate dehydrogenase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract This study reports the use of lactate dehydrogenase release to monitor changes in culture viability in flask culture and fixed bed, porosphere bioreactor systems. Lactate dehydrogenase release shows good agreement with increase in non-viable cell numbers and decline in glucose utilisation in flask cultures. Studies with the immobilised system show that lactate dehydrogenase release can detect loss of viability which is not always indicated by a decrease in glucose utilisation. The data show that culture viability in a repeated-feed-and-harvest system is influenced markedly by both a) the medium change regime itself and b) the use of an immobilised bioreactor compared to a flask system for the same medium change regime.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00365494
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    Paper (German National Licenses)
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  • 4
    Electronic Resource
    Electronic Resource
    Fixed bed porous glass sphere (porosphere) bioreactors for animal cells (1988)
    Springer
    Cytotechnology 1 (1988), S. 339-346 
    Details
    ISSN: 1573-0778
    Keywords: fixed bed ; immobilisation ; perfusion ; scale-up of cell cultures
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Process intensity of fixed bed glass sphere culture systems is increased considerably by replacing solid glass spheres with open pore glass spheres. This technique demonstrates the possibility of having a system capable of both volumetric and cell density scale up and being suitable for substrate attached and suspension cells. The yields achieved for a number of attached cell lines (approximately 107/ml) demonstrate an increase approaching one order of magnitude over solid glass spheres (approximately 106/ml). Also suspension cells were successfully entrapped in the open pore structure with similar yields.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00365079
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    Paper (German National Licenses)
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