Production of recombinant antibody by rCHO cells in bioreactor systems

Abstract

The increasing needs for therapeutic proteins derived from mammalian cells such as recombinant antibody has led to many developments in the area of animal cell technology. In this study, physiological and cultural characteristics of recombinant CHO cells for the production of recombinant antibody was investigated and applied to bioreactor systems for the development of the high productive culture of recombinant production mainly focused on depth filter perfusion culture. In hyper-osmotic condition, Cell growth was inhibited, otherwise, specific productivities was increased. Cellular growth rate and cell density integral was increased, otherwise, specific productivities were decreased, when Glycine betaine was added. In DFPS, near 400 mOsm/kg, no significant harmful inhibition and antibody productivity decrease was occurred for future fortified medium design. Addition of Sodium butyrate (1,2,4,8 mM) had a severe cytotoxicity on rCHO cells. However, Specific Productivity was increased sharply. In the aspect of perfusion culture for higher productivity, Addition of sodium butyrate may lead unstable operation of long term culture. Change of culture temperature had an effect on cell growth, longevity, and productivity. For temperature strategies for high productivity, first, temperature shift from 37 ℃ to 33 ℃ at 120 hour from innoculm was 28% higher antibody concentration than that of 37 ℃ stable control experiment. Second, temperature oscillation between 37 ℃ and 33 ℃ for a day and for other day could get 47%, 39% higher antibody concentration than that of 37 ℃ stable control experiment, respectively. Medium component screening for enriched medium development showed several candidates for $rCHO CS^*13-1.00$. The $rCHO 26^*-320$ cells were successfully cultivated in a depth filter perfusion system, showing high cell density, high antibody production rate, and stable long-term culture. In the DFPS, the volumetric productivities were 43 and 53 times higher th...