Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 333860
Title Effect of feed and bleed rate on Hybridoma cells in an acoustic perfusion bioreactor: Part 1. Cell density, viability and cell-cycle distribution
Author(s) Dalm, M.C.F.; Cuijten, S.M.R.; Grunsven, W.M.J. van; Tramper, J.; Martens, D.E.
Source Biotechnology and Bioengineering 88 (2004)5. - ISSN 0006-3592 - p. 547 - 557.
DOI https://doi.org/10.1002/bit.20287
Department(s) Bioprocess Engineering
VLAG
Publication type Refereed Article in a scientific journal
Publication year 2004
Keyword(s) continuous suspension-culture - hamster ovary cells - protein-free medium - serum-free medium - antibody productivity - mammalian-cells - interferon-gamma - batch culture - growth-rate - retention
Abstract For the development of optimal perfusion processes the effect of the feed and bleed rate on cell growth in a perfusion bioreactor was studied. The viable-cell density, viability, growth, death, and lysis rate and cell-cycle distribution of a hybridoma cell line producing an IgG1 were studied over a range of specific feed and bleed rates. It was found that the feed and bleed rates applied in the different cultures could be divided into two regions based on the viable-cell density and cell-cycle distribution. The cultures in the first region, low feed rates (0.5 and 1.0 d(-1)) combined with low bleed rates (0.05 and 0.10 d(-1)), were nutrient-limited, as an increase in the feed rate resulted in an increase in the viable-cell density. The cultures in the second region, high feed and bleed rates, were nonnutrient-limited. In this region the viable-cell density decreased more or less linearly with an increase in the bleed rate and was independent of the feed rate. This suggests that the cells were limited by a cell-related factor. Comparison of Trypan-blue dye-exclusion measurements and lactate-dehydrogenase activity measurements revealed that cell lysis was not negligible in this bioreactor set-up. Therefore, lactate-dehydrogenase activity measurements were essential to measure the death rate accurately. The specific growth rate was nearly constant for all tested conditions. The viability increased with an increase of the bleed rate and was independent of the feed rate. Furthermore, the specific productivity of monoclonal antibody was constant under all tested conditions. For the optimal design of a perfusion process it should first be established whether viability is an important parameter. If not, a bleed rate as low as possible should be chosen. If low viabilities are to be avoided, the bleed rate chosen should be higher, with the value depending on the desired viability. Next, the feed rate should be set at such a rate that the cells are just in the nonnutrient-limited region. (C) 2004 Wiley Periodicals, Inc.
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