Monoclonal antibody productivity and the metabolic pattern of perfusion cultures under varying oxygen tensions

Abstract

The metabolic pattern and cell culture kinetics of high-cell-density perfusion cultures were compared under two different oxygen transfer conditions: oxygen limiting and not limiting. When oxygen was a limiting factor during perfusion culture, both specific glucose uptake and lactate production rates increased, compared to non-oxygen-limited condition, by about 60% and 30%, respectively. The specific glutamine uptake rate under oxygen-limited conditions was almost 4.0 times higher than that under non-oxygen-limited conditions. The activity of lactate dehydrogenase (LDH) released into the medium by the dead cells can be used as an indicator for the metabolic and physiological conditions related to oxygen limitation. There was a 3.2 times higher specific rate of LDH activity released by dead cells in oxygen-limited cultures than those in non-oxygen-limited cultures. The specific production rate of monoclonal antibody was not significantly affected by the oxygen transfer conditions during the rapid cell growth period, but it rapidly increased toward the end of perfusion cultures. The higher perfusion rate may have limited further cell growth during high-cell-density perfusion culture, because cell damage was caused by the hydrodynamic shear within a hollow fiber microfiltration cartridge installed to withdraw the spent medium and the waste metabolites.

Apoptosis-resistant E1B-19K-expressing NS/0 myeloma cells exhibit increased viability and chimeric antibody productivity under perfusion culture conditions

“…The significant reduction in the growth rate of E1B-19K-expressing cells was also reflected in an increase in the perfusion to batch q MAb ratio. Indeed, many mammalian cell lines cultivated in perfusion exhibit an increase in specific MAb productivity under such slow growth conditions (Al-Rubeai et al, 1992;Banik and Heath, 1995;Batt et al, 1990;de la Broise et al, 1991de la Broise et al, , 1992Johnson et al, 1996;Hansen et al, 1993;Mercille et al, 1994a-c;Shi et al, 1993;Tokashiki and Takamatsu, 1993;Trampler et al, 1994). The q MAb has been shown to be cell cycle dependent and optimum in the G 1 (Byars and Kidson, 1970;Cazzador and Mariani, 1993;Garatun-Tjeldsto et al, 1976;Linardos et al, 1992;Mercille and Massie, 1998;Ramirez and Mutharasan, 1990;Richieri et al, 1991;Suzuki and Ollis, 1989) or G 2 /M phase (Cherlet et al, 1995;Kromenaker and Srienc, 1991;Mercille and Massie, 1998;Watanabe et al, 1973).…”

Heterogeneous conditions in dissolved oxygen affect N‐glycosylation but not productivity of a monoclonal antibody in hybridoma cultures

“…On the other hand, limiting DOT (below 2%) increases hybridoma viability (Miller at al., 1988;Ozturk and Palsson, 1990). The specific monoclonal antibody (MAb) production rate has been shown to remain constant throughout a wide range of DOT (Ozturk and Palsson, 1991;Shi et al, 1993), although an optimum DOT between 10% and 80% has been reported for MAb concentration (Ozturk and Palsson, 1991). In contrast to the relatively abundant reports of the effect of constant DOT on hybridoma metabolism, physiology, and culture performance, no information exists for the case of fluctuating DOT.…”

“…In hybridomas, which exhibit a high glycolytic activity even in aerobic conditions, oxygen determines the consumption rates of carbon and energy sources (glucose and glutamine) and the production of toxic metabolites such as lactate and ammonia (Miller et al, 1988;Ozturk and Palsson, 1991;Shi et al, 1993). The intracellular redox state is also a function of DOT (Hevehan and Miller, 1999;Zupke et al, 1995).…”

Inducible expression of Bcl‐XL restricts apoptosis resistance to the antibody secretion phase in hybridoma cultures

“…Extensive studies on the growth and antibody production characteristics of hybridomas have revealed that antibody secretion is not growth-associated, but rather that these cells exhibit higher speci®c productivities during the late phase of batch cultures (De la Boise et al, 1991;Flickenger et al, 1990;Shi et al, 1993). However, it is now well established that the conditions of nutrient deprivation observed at the late stage of batch cultures can induce apoptotic death of B-cell hybridomas and thus limit antibody productivity (Duval et al, 1990;Mercille et al, 1994b;Perreault and Lemieux, 1993b;Vomastek and Franek, 1993).…”