Biotechnology and Bioengineering, Vol.48, No.2, 118-122, 1995
Overexpression of Bcl-2, Apoptosis Suppressing Gene - Prolonged Viable Culture Period of Hybridoma and Enhanced Antibody-Production
Human bcl-2 DNA was introduced into mouse hybridoma 2E3 cells and expressed at a high level by using BCMGSneo vector, which reportedly amplifies as multiple copies in the cells independently of their chromosomes. The high expression of bcl-2 in BCMGSneo-bcl-2 transfectants was confirmed by western blotting. In batch cultures, the overexpression of bcl-2 raised the maximum viable cell density by 45%, delayed the initiation of apoptosis by 2 days, and prolonged the viable culture period by 4 days. The delayed initiation of apoptosis was detected by emergence of the ladder pattern on DNA electrophoresis and increase of the dead cell number. The bcl-2 transfectants produced IgG(1) fourfold per batch culture in comparison with 2E3 cells transfected with BCMGSneo but not with bcl-2 : a little less than twofold due to the improved survival of the cells and more than twofold due to the enhanced IgG(1) production rate per cell of the bcl-2 transfectants. The method to engineer hybridoma cells genetically with bcl-2 using BCMGSneo vector for increasing viability and productivity would be widely applied for improving antibody productivity of hybridoma cultures.