Mathematical descriptions of hybridoma culture kinetics: I. Initial metabolic rates

Abstract

The lack of quantitative descriptions of mammalian culture kinetics limits the ability to optimally design and control cell culture bioreactors. This limitation is ad dressed by developing mathematical equations relating the initial growth rate and the antibody productivity of the hybridoma cell line, CRL-1606, to its environmental state. This initial rate approach, in contrast with steady state analysis of chemostat cultures, approximates steady state behavior, since the rates were measured over brief time intervals and at low cell concentrations (< 50,000 cells/mL). The advantage of this approach is that it is much faster than the chemostat approach.An equation for the growth rate was developed that superimposed Monod equations in serum and glutamine with "noncompetitive" -type inhibition constants were inversely proportional to the lactate and ammonium concentrations. The Monod constant is critical for relating initial, low cell level culture states to other states.Lactate was found to be the only environmental parameter that significantly inhibited antifibronectin monoclonal antibody production by the CRL-1606 hybridomas. Volumetric productivity was strictly related to culture viability, which was observed to decline at growth rates below 0.02 h(-1). Lactate was also found to significantly inhibit ammonium production.

Effect of serum-free and serum-containing medium on cellular levels of ER-based proteins in various mouse hybridoma cell lines

“…Strategies to enhance MAB yield have extensively involved medium formulation (Glacken et al, 1988;Glassy et al, 1988;Merten and Litwin, 1991;Reuveny et al, 1985;Velez et al, 1986), modes of cell propagation (Altshuler et al, 1986;Boraston et al, 1984;Fazekas de St. Groth, 1983;Reuveny et al, 1986), and the design of high performance reactor configurations (Altshuler et al, 1986;Arathoon and Birch, 1986;Brunt, 1986;Hu et al, 1985;Posillico, 1986). Most of these procedures have focused on demonstrating the feasibility of the process (Leno et al, 1992a).…”

Establishment of an apoptosis-resistant and growth-controllable cell line by transfecting with inducible antisense c-Jun gene

“…Therefore, preventing cell death that starts in the late logarithmic growth phase (Vomastek and Franek, 1993) and maintaining the cells viable in batch culture for longer time periods should increase protein production of the culture. The cells die due to depletion of nutrients such as amino acids (Franek and Chladkova-Sramkova, 1995) and glucose (Mercille and Massie, 1994;Singh et al, 1994), limitation of growth factors such as serum components (Singh et al, 1994), or accumulation of toxic metabolites (Glacken et al, 1988;Tohyama et al, 1990). The overgrowth inevitably leads to some of these cell death inducing conditions.…”

Metabolic‐flux analysis of continuously cultured hybridoma cells using 13CO2 mass spectrometry in combination with 13C‐lactate nuclear magnetic resonance spectroscopy and metabolite balancing

“…Apparently, they take up higher amounts of glucose and glutamine than they actually require for energy and biosynthesis. The accumulation of these waste products inhibits cell growth, reduces cell-specific production, and affects product quality (Anderson and Goochee, 1995;Glacken et al, 1988;Kimura and Miller, 1996;Ozturk et al, 1992).…”