Application of Anti-Death Proteins to Mammalian Cell Culture

Bruno Figueroa Jr, Tina M. Sauerwald, J. Marie Hardwick, and Michael J. Betenbaugh
Johns Hopkins University
3400 North Charles Street
Baltimore, MD  21218
bruno@jhu.edu
(410) 516-5336

Mammalian cells undergo apoptosis due to various stimuli common in large-scale protein production schemes. In our research, CHO and BHK cells have been genetically engineered with anti-death members of the Bcl-2 family, bcl-2 and bcl-xL. Both CHO and BHK modified cells were subjected to two model apoptotic stimuli, Sindbis virus and serum deprivation.  Apoptosis was delayed but not completely inhibited by the overexpression of the anti-death genes. Biological changes occurring within the cell appear to alter the anti-death proteins’ effectiveness to inhibit apoptosis.