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Enhancement of productivity of recombinant alpha-amidating enzyme by low temperature culture.
Furukawa K
,
Ohsuye K
.
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We have produced a recombinant C-terminal alpha-amidating enzyme (799BglIIalpha-AE) derived from Xenopus laevis by culturing a CHO cell line named 3mu-1S. Recently, we demonstrated that culturing 3mu-1S cells at a temperature below 37 degrees C led to the following phenomena: inhibited cell growth with high viability, enhanced cellular productivity (maximally at 32 degrees C), and suppressed medium consumption and release of impurities from the cells. Therefore, it is suggested that the 799BglIIalpha-AE production will be increased by culturing a sufficient number of the cells at a low temperature (especially at 32 degrees C). To assess this effect on batch and perfusion cultures, the culture temperature was shifted from 37 to 32 degrees C in the mid-exponential phase in the case of batch culture and from 37 to 34 degrees C when the cell density became high enough in the case of perfusion culture. Application of the low temperature culture to batch and perfusion cultures was effective in comparison with the culture at 37 degrees C: the productivity per medium and the productivity per time were increased severalfold with enhanced cellular productivity at a low culture temperature. The low temperature culture also increased the relative content of 799BglIIalpha-AE in the supernatant and reduced the glucose consumption. The method presented here would contribute to production of bioactive proteins using other recombinant cell lines.
Borth,
Growth and production kinetics of human x mouse and mouse hybridoma cells at reduced temperature and serum content.
1992, Pubmed
Borth,
Growth and production kinetics of human x mouse and mouse hybridoma cells at reduced temperature and serum content.
1992,
Pubmed
Bradbury,
Mechanism of C-terminal amide formation by pituitary enzymes.
1982,
Pubmed
Chevrier-Miller,
Low temperature enhancement of reporter genes expression directed by human immunodeficiency virus type 1 long terminal repeat.
1996,
Pubmed
Chuppa,
Fermentor temperature as a tool for control of high-density perfusion cultures of mammalian cells.
1997,
Pubmed
Eipper,
Identification in pituitary tissue of a peptide alpha-amidation activity that acts on glycine-extended peptides and requires molecular oxygen, copper, and ascorbic acid.
1983,
Pubmed
Furukawa,
Effect of culture temperature on a recombinant CHO cell line producing a C-terminal α-amidating enzyme.
1998,
Pubmed
,
Xenbase
Giard,
Examination of parameters affecting human interferon production with microcarrier-grown fibroblast cells.
1980,
Pubmed
Giard,
Effect of temperature on the production of human fibroblast interferon.
1982,
Pubmed
Kato,
Two enzymes concerned in peptide hormone alpha-amidation are synthesized from a single mRNA.
1990,
Pubmed
Kojima,
Enhanced production of interferon by temperature shift-down from 37 C to 25 C in rabbit cell cultures stimulated with Newcastle disease virus.
1974,
Pubmed
Mizuno,
Cloning and sequence of cDNA encoding a peptide C-terminal alpha-amidating enzyme from Xenopus laevis.
1987,
Pubmed
,
Xenbase
Mizuno,
Peptide C-terminal alpha-amidating enzyme purified to homogeneity from Xenopus laevis skin.
1986,
Pubmed
,
Xenbase
Ohsuye,
Cloning of cDNA encoding a new peptide C-terminal alpha-amidating enzyme having a putative membrane-spanning domain from Xenopus laevis skin.
1988,
Pubmed
,
Xenbase
Reuveny,
Factors affecting cell growth and monoclonal antibody production in stirred reactors.
1986,
Pubmed
Sureshkumar,
The influence of temperature on a mouse-mouse hybridoma growth and monoclonal antibody production.
1991,
Pubmed
Takagi,
Comparison of the optimal culture conditions for cell growth and tissue plasminogen activator production by human embryo lung cells on microcarriers.
1994,
Pubmed
Vilcek,
Stabilization of interferon messenger RNA activity by treatment of cells with metabolic inhibitors and lowering of the incubation temperature.
1973,
Pubmed
Weidemann,
Low temperature cultivation--a step towards process optimisation.
1994,
Pubmed
