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  • 1
    Electronic Resource
    Electronic Resource
    Membrane chromatography - an integrative concept in the downstream processing of proteins (1995)
    s.l. : American Chemical Society
    Biotechnology progress 11 (1995), S. 357-367 
    Details
    ISSN: 1520-6033
    Source: ACS Legacy Archives
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1021/bp00034a001
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    An approach to integrated antibody production: Coupling of fluidized bed cultivation and fluidized bed adsorption (1996)
    Springer
    Bioprocess and biosystems engineering 15 (1996), S. 21-29 
    Details
    ISSN: 1432-0797
    Source: Springer Online Journal Archives 1860-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Continuous culture may be an efficient way of producing proteins which are susceptible to secondary processing in the course of a fermentation process. Short residence times in these systems support the production of correctly assembled proteins by avoiding substrate limitations and product inhibitions and also minimize the contact of sensitive bioproducts with degrading enzymes. Thus products of increased stability and integrity are obtained from continuous processes. The downstream process following continuous culture has to be adapted to the specific conditions of continuous fermentations, e.g. large liquid volumes and diluted process solutions. In this paper an approach is shown how a fluidized bed adsorption as first recovery operation may be coupled directly to a continuous production. Immobilized hybridoma cells are cultivated in porous glass microcarriers in a continuous fluidized bed process, the cell containing harvest is purified by fluidized bed adsorption using an agarose based cation exchange matrix. By this coupled mode of operation the large biomass containing harvest volume resulting from the continuous cultivation may be applied directly to a fluidized chromatographic matrix without prior clarification, leading to a particle free and initially purified product solution of reduced volume. In an experimental setup a bench-scale fluidized bed bioreactor of 25 ml carrier volume was coupled to a fluidized bed adsorption column operated with 300 ml of adsorbent. This configuration yielded up to 20 mg of monoclonal antibody per day in a cell free solution at fourfold concentration and fivefold purification. The process was run for more than three weeks with consistent product output.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00435523
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    Paper (German National Licenses)
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  • 3
    Electronic Resource
    Electronic Resource
    Fed-batch production of recombinant human calcitonin precursor fusion protein using Staphylococcus carnosus as an expression-secretion system (2000)
    Springer
    Applied microbiology and biotechnology 54 (2000), S. 361-369 
    Details
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A pH-auxostatic fed-batch process was developed for the secretory production of a fusion protein consisting of the pro-part of Staphylococcus hyicus lipase and two synthetic human calcitonin (hCT) precursor repeats under the control of a xylose-inducible promotor from Staphylococcus xylosus. Using glycerol as the energy source and pH-controlled addition of yeast extract resulted in the production of 2000 mg l−1 of the fusion protein (420 mg l−1 of the recombinant hCT precursor) within 14 h, reaching 45 g l−1 cell dry mass with Staphylococcus carnosus in a stirred-tank reactor. Product titer and space-time yield (30 mg calcitonin precursor l−1 h−1) were thus improved by a factor of 2, and 4.5, respectively, compared to Escherichia coli expression-secretion systems for the production of calcitonin precursors. Two hundred grams of the fusion protein was secreted by the recombinant S. carnosus on a 150-l scale (scale-up factor of 50) with a minimum use of technical-grade yeast extract (40 mg fusion protein g−1 yeast extract).
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002530000406
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  • 4
    Electronic Resource
    Electronic Resource
    The influence of dissolved oxygen tension on the metabolic activity of an immobilized hybridoma population (1993)
    Springer
    Cytotechnology 13 (1993), S. 29-39 
    Details
    ISSN: 1573-0778
    Keywords: collagen microspheres ; DOT influence ; fluidized bed bioreactor ; monoclonal antibody ; perfusion culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract In the study presented here a laboratory scale (150 ml) fluidized bed bioreactor was used as a tool for making kinetic measurements on the production of monoclonal antibodies (MAb) with a hybridoma cell line. We determined the influence of dissolved oxygen tension (DOT) on the metabolic activity of a hybridoma population immobilized in macroporous collagen microspheres. The data obtained showed a reduction of the metabolic activity of the immobilized population at reduced DOT, the total number of immobilized cells, however, remained constant. At decreasing DOT an increasing lactate yield from glucose at reduced glutamine consumption was noticed, indicating a shift in the pattern of substrate usage. A mathematical description of maintenance metabolism was formulated and the parameters of growth and maintenance requirements were calculated. A growth associated MAb production was determined under the conditions applied leading to space time yields of 225 mg MAb per liter of total reactor volume and day.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00749973
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    Paper (German National Licenses)
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  • 5
    Electronic Resource
    Electronic Resource
    An approach to integrated antibody production: Coupling of fluidized bed cultivation and fluidized bed adsorption (1996)
    Springer
    Bioprocess engineering 15 (1996), S. 21-29 
    Details
    ISSN: 0178-515X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract  Continuous culture may be an efficient way of producing proteins which are susceptible to secondary processing in the course of a fermentation process. Short residence times in these systems support the production of correctly assembled proteins by avoiding substrate limitations and product inhibitions and also minimize the contact of sensitive bioproducts with degrading enzymes. Thus products of increased stability and integrity are obtained from continuous processes. The downstream process following continuous culture has to be adapted to the specific conditions of continuous fermentations, e.g. large liquid volumes and diluted process solutions. In this paper an approach is shown how a fluidized bed adsorption as first recovery operation may be coupled directly to a continuous production. Immobilized hybridoma cells are cultivated in porous glass microcarriers in a continuous fluidized bed process, the cell containing harvest is purified by fluidized bed adsorption using an agarose based cation exchange matrix. By this coupled mode of operation the large biomass containing harvest volume resulting from the continuous cultivation may be applied directly to a fluidized chromatographic matrix without prior clarification, leading to a particle free and initially purified product solution of reduced volume. In an experimental setup a bench-scale fluidized bed bioreactor of 25 ml carrier volume was coupled to a fluidized bed adsorption column operated with 300 ml of adsorbent. This configuration yielded up to 20 mg of monoclonal antibody per day in a cell free solution at fourfold concentration and fivefold purification. The process was run for more than three weeks with consistent product output.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00435523
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    Paper (German National Licenses)
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  • 6
    Electronic Resource
    Electronic Resource
    The influence of particle size distribution and operating conditions on the adsorption performance in fluidized beds (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 55 (1997), S. 54-64 
    Details
    ISSN: 0006-3592
    Keywords: fluidized-bed adsorption ; dispersion ; particle diameter ; bed height ; frontal adsorption ; mass transport ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The influence of matrix properties and operating conditions on the performance in fluidized-bed adsorption has been studied using Streamline diethyl-aminoethyl (DEAE), an ion exchange matrix based on quartz-weighted agarose, and bovine serum albumin (BSA) as a model protein. Three different particle size fractions (120-160 μm, 120-300 μm, and 250-300 μm) were investigated. Dispersion in the liquid phase was reduced when particles with a wide size distribution were fluidized compared to narrow particle size distributions. When the mean particle diameter was reduced, the breakthrough capacities during frontal adsorption were enlarged due to a shorter diffusion path length within the matrix. At small particle diameters the effect of film mass transfer became more relevant to the adsorption performance in comparison to larger particles. Therefore matrices designed for fluidized-bed adsorption should have small particle diameter and increased mean particle density to ensure small diffusion path length in the particle and a high interstitial velocity to improve film mass transfer. Studies on the influence of sedimented matrix height on axial mixing showed an increased Bodenstein number with increasing bed length. Higher breakthrough capacities were also found for longer adsorbent beds due to reduced dispersion and improved fluid and particle side mass transfer. With increasing bed height the influence of flow rate on breakthrough capacity was reduced. For a settled bed height of 50 cm breakthrough capacities of 80% of the equilibrium capacity for flow rates varying from 3 to 9 cm/min could be achieved. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 55: 54-64, 1997.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
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    Paper (German National Licenses)
  • 7
    Electronic Resource
    Electronic Resource
    Pilot scale processing of detergent-based aqueous two-phase systems (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 55 (1997), S. 339-347 
    Details
    ISSN: 0006-3592
    Keywords: detergent based aqueous two phase systems ; phase separation by sedimentation ; phase separation by centrifugation ; separation of detergent based aqueous phase systems ; biomass influence ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Detergent based aqueous two-phase systems have several specific properties, e.g., extreme small density differences between the two liquid phases (0.003-0.005 g/cm3), low interfacial tensions (5-10 μN/m) and complex rheological behavior of the product containing detergent-rich phase, which make processing difficult. We describe the successful separation of these aqueous two-phase systems in the pilot scale (1-20 kg) in the presence and absence of microbial cells, either by settling under gravity or in centrifugal separators. The performance of self-desludging liquid-liquid separators and of a nozzle separator was analyzed in detail to judge large scale application. With a feed rate of 16 L/h, stable operation was possible in the desludging machine. Up to 56 L/h could be processed with very close control of the hydrodynamic balance. In a small nozzle separator, feed rates of 90 L/h could be realized, but the purity of the separated phases and the yield of the top phase was slightly lower than in the liquid-liquid separator. The presence of surface-active components in the feed may alter the separation characteristics of the phase systems significantly. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 55: 339-347, 1997.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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    Paper (German National Licenses)
  • 8
    Electronic Resource
    Electronic Resource
    52. Stofftransportmechanismen in der Fließbettchromatographie und ihre Bedeutung für die Adsorption von Proteinen (1998)
    Weinheim : Wiley-Blackwell
    Chemie Ingenieur Technik - CIT 70 (1998), S. 1100-1101 
    Details
    ISSN: 0009-286X
    Keywords: Chemistry ; Industrial Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/cite.330700956
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    Paper (German National Licenses)
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