Co-expression of bcl-2 and bag-1, apoptosis suppressing genes, prolonged viable culture period of hybridoma and enhanced antibody production |
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Authors: | Satoshi Terada Tomoaki Komatsu Tetsuo Fujita Ain Terakawa Teruyuki Nagamune Shinichi Takayama John C Reed Eiji Suzuki |
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Abstract: | Human bcl-2 and bag-1 DNA were introduced into mouse hybridoma 2E3- O cells and expressed. The expression of bcl-2 in BCMGneo-bcl2
transfectants was confirmed by ELISA and that of bag-1 in pZeo-bag1 was confirmed by western blotting. In batch cultures,
the over-expression of bcl-2 prolonged the culture period by 2 days and co-expression of bcl-2 and bag-1 prolonged the culture
period by 3 days. The delayed increase in the dead cell number in culture of the bcl-2 and bag-1 cotransfectant indicated
the additional antiapoptosis effect of bcl-2 and bag-1 cotransfection in comparison with the bcl-2 only transfection. The
bcl-2 transfectants (2E3O-Bcl2) produced antibody twofold per batch culture in comparison with 2E3-O cells transfected with
BCMGSneo (2E3O-Mock). Enhancement of this MoAb production was due to the improved survival of the cells and was not due to
stimulation of antibody production rate per cell by Bcl-2 expression. And the bcl-2 and bag-1 co-transfectant (2E3O-Bcl2-BAG1)
produced antibody approximately fourfold of 2E3O-Mock per batch culture. Enhancement of this MoAb production was due to the
improved survival of the cells and was partly due to stimulation of MoAb production rate per cell in the non-growing phase
by the cotransfection. The method to engineer hybridoma cells genetically with bcl-2 and bag-1 for increasing viability and
productivity would be widely applied for improving antibody productivity of hybridoma cultures.
This revised version was published online in August 2006 with corrections to the Cover Date. |
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Keywords: | antibody productivity apoptosis BAG-1 Bcl-2 cell survival hybridoma |
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