Increased production of protein from senescent cells. Monoclonal antibodies (mAbs) are emerging as robust therapeutic options in the fight against cancer and autoimmune diseases. The estimated market in 2004 for therapeutic mAbs is more than $7 billion from 16 FDA approved biopharmaceutical products. Kilogram quantities of mAbs are produced on in large-scale bioreactors from traditional hybridoma cultures or genetically engineered mammalian cells. Bioprocessing of mAbs is a prohibitively expensive endeavor, costing nearly $10,000 per gram. There is a tremendous need for technologies that can make bioprocessing of mAbs easier and reduce production costs to make mAb therapies more affordable. Our company has developed a novel technology that produces premature senescence of cells in culture resulting in a 30-fold enhancement in the production of secreted proteins. The major goal of the proposed Phase I project is to translate this senescence technology into commercially relevant hybridoma cells. Specifically, plasmid or retroviral vectors with tetracycline-inducible promoters driving expression of cyclin-dependent kinase inhibitors are introduced into hybridoma cells. Following antibiotic selection, premature-senescence is tested in populations of induced-transformed cells, as measured by relative cell growth, DNA cell cycle analysis, and expression of senescence-associated biomarkers. Senescence-competent hybridoma cell lines are isolated by limiting dilution and tested individually for enhanced production of immunoglobulin. Future Phase I project goals are to transfer senescent-competent cell lines developed in this Phase I project into commercial-scale bioreactors to assess mAb production on a large scale. We intend to license this technology to biopharmaceutical companies and contract manufacturing laboratories to reduce their cost in bioprocessing of therapeutic proteins from mammalian cells.
