CELL GROWTH ARREST BY NUCLEOTIDES AS A TOOL FOR IMPROVED PRODUCTION OF BIOPHARMACEUT1CALS

摘要

Arresting cell growth and thus decreasing cell division potentially lessens the chance for genetic drift in the cell population: this would be of utmost importance for the consistent production of high qualitx products. Cell growth arrest of two industrially relevant animal cell lines (CHO and BKK cells) has been successfully achieved b) the overexpression of a cyclin-dependen: kinase inhibitor (p27) and of a tumor suppressor gene (1RF-1) (genetic strategy), punnes addition (chemical strategy) and bythe depletion of essential nutrients from the culture medium (environmental strategy). Adenosine and AMP addition, as well as the ovcrexpression of p27 led to a 3 to 4-fold increase in the specific productivity of two recombinant proteins: the secretedform of the human placental alkaline phosphatase (SEAP) and the coagulation Factor VUI. AMP addition showed to be the most promising tool for biopharmacemicals production; fine tuning of drug addition was required. All the tested cell growth arrest strategies interfered not only with cell cycle but also with cell metabolism. A careful look onto cell cycle distribution in the different scenarios created, shows whether it is important to consider recombinant protein expression dependency upon cell cycle in process optimization and operation strategies.