Cell cycle kinetics and monoclonal antibody productivity of hybridoma cells during perfusion culture

The flow‐cytometric (FCM) analysis of bivariate DNA/lgG distributions has been conducted to study the cell cycle kinetics and monoclonal antibody (MAb) production during perfusion culture of hybridoma cells. Three different perfusion rates were employed to demonstrate the dependency of MAb synthesis and secretion on cell cycle and growth rate. The results showed that, during the rapid growth period of perfusion culture, the level of intracellular igG contents of hybridoma cells changed significantly at each perfusion rate, while the DNA histograms showing cell cycle phases were almost constant. Meanwhile, during the reduced growth period of perfusion culture, the fraction of cells in the S phase decreased, and the fraction cells in the G1/G0 phase increased with decreasing growth rate. The fraction of cells in the G2/M phase was relatively constant during the whole period of perfusion culture. Positive correlation was found between mean intracellular IgG contents and the specific MAb production rate, suggesting that the deletion of intracellular IgG contents by a flow cytometer could be used as a good indicator for the prediction of changes in specific MAb productivity following manipulation of the culture condition. © 1994 John Wiley & Sons, Inc.

[1]  B. Palsson,et al.  Immobilization can improve the stability of hybridoma antibody productivity in serum‐free media , 1990, Biotechnology and bioengineering.

[2]  D S Kompala,et al.  Inclined Sedimentation for Selective Retention of Viable Hybridomas in a Continuous Suspension Bioreactor , 1990, Biotechnology progress.

[3]  J. Hamlin,et al.  Animal cell cycle. , 1978, Annual review of biochemistry.

[4]  M. Gray,et al.  Effect of temperature on hybridoma cell cycle and MAb production , 1992, Biotechnology and bioengineering.

[5]  C. Y. Yang,et al.  Preparation and characterization of monoclonal antibodies to cephalexin-synthesizing enzyme from Acetobacter turbidans. , 1988, Hybridoma.

[6]  S. Reuveny,et al.  Comparison of cell propagation methods for their effect on monoclonal antibody yield in fermentors , 1986 .

[7]  N. Kalogerakis,et al.  Cell cycle model for growth rate and death rate in continuous suspension hybridoma cultures , 1992, Biotechnology and bioengineering.

[8]  B. Palsson,et al.  Population balance between producing and nonproducing hybridoma clones is very sensitive to serum level, state of inoculum, and medium composition. , 1992, Biotechnology and Bioengineering.

[9]  R. Mutharasan,et al.  Cell cycle‐ and growth phase‐dependent variations in size distribution, antibody productivity, and oxygen demand in hybridoma cultures , 1990, Biotechnology and bioengineering.

[10]  D F Ollis,et al.  A flow‐cytometric analysis of hybridoma growth and monoclonal antibody production , 1990, Biotechnology and bioengineering.

[11]  J. Gray,et al.  Techniques in Cell Cycle Analysis , 1987, Humana Press.

[12]  D. Needham,et al.  A physical characterization of GAP A3 hybridoma cells: Morphology, geometry, and mechanical properties , 1991, Biotechnology and bioengineering.

[13]  M. Cornblath,et al.  Reciprocal regulation of glucose and glutamine utilization by cultured human diploid fibroblasts , 1978, Journal of cellular physiology.

[14]  J. Gabriels.  A rapid high performance liquid chromatography technique for monitoring amino acid utilization in cell culture , 1984 .

[15]  E. C. Beuvery,et al.  Effect of dilution rate on growth, productivity, cell cycle and size, and shear sensitivity of a hybridoma cell in a continuous culture , 1993, Biotechnology and bioengineering.

[16]  B. Palsson,et al.  Loss of antibody productivity during long-term cultivation of a hybridoma cell line in low serum and serum-free media. , 1990, Hybridoma.