A structured model for monoclonal antibody synthesis in exponentially growing and stationary phase hybridoma cells

An initial structured unsegregated kinetic model describing monoclonal antibody synthesis by a murine hybridoma cell line (9.2.27) grown in 1 liter batch cultures is described. The model is based on the intracellular balances of the heavy and light chain coding mRNAs, the intracellular balances of heavy and light chains and the description of the kinetics of heavy and light chain assembly. Model parameters were varied with specific growth rate in order to account for changes in the rates of antibody synthesis and secretion with entrance of the cells from the exponential into the stationary phase of growth. The parameters were varied based. on experimental data obtained in our laboratory on the variation of total cellular RNA content and the half‐lives of heavy (H) and light (L) chain mRNAs with specific growth rate, and data from other investigators on immunoglobulin synthesis and secretion. The model successfully predicts the experimentally observed decrease in the intracellular heavy and light chain mRNA levels with entrance of 9.2.27 cells from the exponential into the stationary phase of growth, as well as the extracellular accumulation of antibody (IgG2a) during batch culture.

[1]  John Paul,et al.  Tissue culture: Methods and applications: Ed. by Paul F. Kruse, Jr and M. K. Patterson, Jr. 1973. New York and London: Academic Press. Pp. xxvii and 868; 264 text figs. £10.15. , 1974 .

[2]  S. Tonegawa,et al.  Transcription of mouse kappa chain genes: implications for allelic exclusion. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[3]  B. Palsson,et al.  On the orders of magnitude of epigenic dynamics and monoclonal antibody production , 1989 .

[4]  D. Nuss,et al.  Variation in the relative synthesis of immunoglobulin G and non-immunoglobulin G proteins in cultured MPC-11 cells with changes in the overall rate of polypeptide chain initiation and elongation. , 1976, Journal of molecular biology.

[5]  M. A. Bohn,et al.  Determination of specific monoclonal antibody secretion rate during very slow hybridoma growth , 1990 .

[6]  D. Galloway,et al.  Production and characterization of monoclonal antibody to a melanoma specific glycoprotein. , 1981, Hybridoma.

[7]  J. Kearney,et al.  Posttranslational association of immunoglobulin heavy chain binding protein with nascent heavy chains in nonsecreting and secreting hybridomas , 1986, The Journal of cell biology.

[8]  H. Moses,et al.  Effect of cell proliferation on levels and diversity of poly(A)-containing mRNA , 1976, Cell.

[9]  U. Schibler,et al.  The synthesis and processing of the messenger RNAs specifying heavy and light chain immunoglobulins in MPC-11 cells , 1978, Cell.

[10]  R F Geoghegan,et al.  Kinetic modelling of hybridoma cell growth and immunoglobulin production in a large-scale suspension culture. , 1988, Biotechnology and bioengineering.

[11]  H. Hiatt,et al.  Actinomycin D: An Effect on Rat Liver Homogenates Unrelated to Its Action on RNA Synthesis , 1964, Science.

[12]  R. Freedman,et al.  Protein disulphide-isomerase and the formation of native disulphide bonds. , 1984, Biochemical Society transactions.

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

[14]  M. Scharff,et al.  Synthesis, assembly and secretion of gamma-globulin by mouse myeloma cells. V. Balanced and unbalanced synthesis of heavy and light chains by IgG-producing tumors and cell lines. , 1973, Journal of immunology.

[15]  C. Milstein,et al.  Stability of cytoplasmic ribonucleic acid in a mouse myeloma: estimation of the half-life of the messenger RNA coding for an immunoglobulin light chain. , 1974, Journal of molecular biology.

[16]  P. Langridge,et al.  Extraction of nucleic acids from agarose gels. , 1980, Analytical biochemistry.

[17]  J. Kearney,et al.  Organization and expression of immunoglobulin genes in fetal liver hybridomas. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[18]  D F Ollis,et al.  Enhanced Antibody Production at Slowed Growth Rates: Experimental Demonstration and a Simple Structured Model , 1990, Biotechnology progress.

[19]  C. Milstein,et al.  Variability of Interchain Binding of Immunoglobulins: Interchain Bridges of Mouse IgG2a and IgG2b , 1970, Nature.

[20]  J. Rothman,et al.  Peptide binding and release by proteins implicated as catalysts of protein assembly. , 1989, Science.

[21]  M. Wabl,et al.  Immunoglobulin mRNA stability varies during B lymphocyte differentiation. , 1988, The EMBO journal.

[22]  H. Lodish.  Molecular Cell Biology , 1986 .

[23]  R. Stevens.  Translational Control of Antibody Synthesis , 1974 .

[24]  R. G. Krueger,et al.  The effect of actinomycin D and 5-azacytidine on macromolecular synthesis in murine myeloma tumor cells. , 1972, Journal of immunology.

[25]  H. Green,et al.  Changes in RNA in relation to growth of the fibroblast: II. The lifetime of mRNA, rRNA, and tRNA in resting and growing cells , 1974 .

[26]  R. Perry,et al.  Lifetime of the messenger RNA's which code for ribosomal proteins in L-cells. , 1971, Biochimica et biophysica acta.

[27]  R. Weinmann,et al.  Mechanism of action of DRB. III. Effect on specific in vitro initiation of transcription. , 1983, Journal of molecular biology.

[28]  M. Percy,et al.  A theoretical model for the covalent assembly of immunoglobulins. Application to the assembly of human immunoglobulin G in vitro. , 1975, The Journal of biological chemistry.

[29]  R. Roth,et al.  Role of disulfide interchange enzyme in immunoglobulin synthesis. , 1981, Biochemistry.

[30]  T. Pihlajaniemi,et al.  Protein disulphide-isomerase activity in various cells synthesizing collagen. , 1983, European journal of biochemistry.

[31]  C. Auffray,et al.  Purification of mouse immunoglobulin heavy-chain messenger RNAs from total myeloma tumor RNA. , 2005, European journal of biochemistry.

[32]  D. Kompala,et al.  A structured kinetic modeling framework for the dynamics of hybridoma growth and monoclonal antibody production in continuous suspension cultures , 1989, Biotechnology and bioengineering.

[33]  J. Rothman.  Polypeptide chain binding proteins: Catalysts of protein folding and related processes in cells , 1989, Cell.

[34]  W M Miller,et al.  A kinetic analysis of hybridoma growth and metabolism in batch and continuous suspension culture: Effect of nutrient concentration, dilution rate, and pH , 2000, Biotechnology and bioengineering.

[35]  Y. Namba,et al.  Immunoglobulin synthesis by cultured mouse myeloma cells. , 1969, Journal of immunology.

[36]  H. Amos,et al.  mRNA half-life measured by use of actinomycin D in animal cells — A caution , 1966 .

[37]  U. Chen.  Anti-IgM antibodies inhibit IgM expression in lipopolysaccharide-stimulated normal murine B-cells: study of RNA metabolism and translation. , 1988, Gene.

[38]  R. Singer,et al.  Biological Sciences: Stability of HeLa Cell mRNA in Actinomycin , 1972, Nature.

[39]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[40]  G. Muscat,et al.  Growth-related changes in specific mRNAs upon lectin activation of human lymphocytes. , 1985, DNA.

[41]  E. Meurs,et al.  Alpha‐interferon inhibits the expression of heavy chain mu messenger RNA in Daudi cells. , 1988, The EMBO journal.

[42]  D. Linzer,et al.  Growth-related changes in specific mRNAs of cultured mouse cells. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[43]  R. Laskov,et al.  Synthesis, assembly and secretion of gamma globulin by mouse myeloma cells. II. Assembly of IgG2b immunoglobulin by MPC 11 tumor and culture cells. , 1971, Journal of molecular biology.

[44]  R. Laskov,et al.  Synthesis and assembly of immunoglobulin polypeptide chains. , 1970, Progress in allergy.

[45]  M. Flickinger,et al.  Large-scale purification of a murine antimelanoma monoclonal antibody , 1986 .

[46]  R. Perry,et al.  Isolation, purification, and properties of mouse heavy-chain immunoglobulin mRNAs. , 1978, Biochemistry.

[47]  P. Leder,et al.  Characterization of light chain and light chain constant region fragment mRNAs in MPC 11 mouse myeloma cells and variants , 1975, Cell.

[48]  R. Perry,et al.  Messenger RNA turnover in mouse L cells. , 1973, Journal of molecular biology.