HYBRIDOMA GROWTH AND ANTIBODY-PRODUCTION AS A FUNCTION OF CELL-DENSITY AND SPECIFIC GROWTH-RATE IN PERFUSION CULTURE

Steady state metabolic parameters for hybridoma cell line H22 were det ermined over a wide range of cell densities and specific growth rates in a filtration based homogeneous perfusion reactor, Operating the rea ctor at perfusion rates of 0.75, 2.0, and 2.9 day(-1) (each at four di fferent specific growth rates), viable cell densities as high as 2 x 1 0(7) cells/mL. were obtained. For the cell line under investigation, t he specific monoclonal antibody production rate was found to be a stro ng function of the viable cell density, increasing with increasing cel l density, In contrast, most of the substrate consumption and product formation rates were strong functions of the specific growth rate. Sub strate metabolism became more efficient at high cell densities and low specific growth rates, The specific rates of metabolite formation and the apparent yields of lactate from glucose and ammonia from glutamin e decreased at low specific growth rates and high cell densities. Whil e the specific oxygen consumption rate was independent of the specific growth rate and cell density, ATP production was more oxidative at lo wer specific growth rate and higher cell density. These observed shift s are strong indications of the production potential of high-density p erfusion culture, (C) 1995 John Wiley & Sons, Inc.