Monoclonal antibody production in murine ascites. II. Production characteristics.

OBJECTIVE To characterize monoclonal antibody production parameters of five hybridoma cell lines in murine ascites for correlation with clinicopathologic changes in mice. METHODS Five hybridoma cell lines were grown in groups of 20 mice. Fourteen days prior to inoculation with 10(6) hybridoma cells, mice were primed with 0.5 ml of pristane given intraperitoneally. Ascites fluid was collected a maximum of three times by abdominal paracentesis; volume was measured and antibody concentration was determined by ELISA for each sample. RESULTS Trends differed among cell lines when comparing ascites volumes and antibody concentrations over time from the first to the third tap. Antibody production was greatest at tap 1 for Groups 2B11 and 2C6D9; tap 2 for Group 3C9; and tap 3 for Groups RMK and 3D6. Total antibody production ranged from 422.90 to 996.64 mg; total ascites fluid volume ranged from 74.2 to 115.7 ml; and mean antibody concentration for taps 1, 2, and 3 ranged from 2.50 to 15.03 mg/ml among cell lines. CONCLUSION Production characteristics were significantly different among hybridoma cell lines. Determination of production characteristics of hybridomas and correlation with clinicopathologic changes in mice may be valuable in making recommendations for managing mice with ascites.

[1]  L. Trudel,et al.  Monoclonal antibody production in murine ascites. I. Clinical and pathologic features. , 1999, Laboratory animal science.

[2]  N. Hoogenraad,et al.  The effect of pre-injection of mice with pristane on ascites tumour formation and monoclonal antibody production. , 1983, Journal of immunological methods.

[3]  H. Fiebig,et al.  In vitro production of monoclonal antibodies in high concentration in a new and easy to handle modular minifermenter. , 1995, Journal of immunological methods.

[4]  K. James,et al.  Human monoclonal antibodies: their potential, problems, and prospects. , 1989, Advances in biotechnological processes.

[5]  Andrew N. Rowan,et al.  Refinement of Monoclonal Antibody Production and Animal Well-being , 1989 .

[6]  I. Ruhdel,et al.  Monoclonal Antibodies: In Vivo and In Vitro Production on a Laboratory Scale, with Consideration of the Legal Aspects of Animal Protection , 1989, ALTEX.

[7]  R. Kurkela,et al.  Pilot-scale production of murine monoclonal antibodies in agitated, ceramic-matrix or hollow-fiber cell culture systems. , 1993, BioTechniques.

[8]  T. Geske,et al.  Laboratory scale production of monoclonal antibodies in a tumbling chamber. , 1995, Journal of immunological methods.

[9]  M. Potter,et al.  The requirement of an adherent cell substratum for the growth of developing plasmacytoma cells in vivo , 1976, The Journal of experimental medicine.

[10]  B. Brodeur,et al.  Parameters affecting ascites tumour formation in mice and monoclonal antibody production. , 1984, Journal of immunological methods.

[11]  R. Gillette.  Alternatives to pristane priming for ascitic fluid and monoclonal antibody production. , 1987, Journal of immunological methods.

[12]  C. Milstein,et al.  [1] Preparation of monoclonal antibodies: Strategies and procedures , 1981 .

[13]  W. Jones,et al.  Monoclonal antibody production by hybridoma growth in Freund's adjuvant primed mice. , 1986, Journal of immunological methods.

[14]  J. Fox,et al.  Institutional Policies and Guidelines on Adjuvants and Antibody Production. , 1995, ILAR journal.

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

[16]  A. Mizrahi.  Monoclonal antibodies: production and application. , 1989, Advances in biotechnological processes.

[17]  Uwe Marx,et al.  Monoclonal Antibody Production , 1997 .

[18]  G. Belfort,et al.  Continuous hybridoma growth and monoclonal antibody production in hollow fiber reactors–separators , 1986, Biotechnology and bioengineering.

[19]  C. Milstein,et al.  An oscillating bubble chamber for laboratory scale production of monoclonal antibodies as an alternative to ascitic tumours. , 1992, Journal of immunological methods.

[20]  Amyx Hl.  Control of animal pain and distress in antibody production and infectious disease studies. , 1987 .

[21]  J. Kalil,et al.  Multiparametric analyses of hybridoma growth on glass cylinders in a packed-bed bioreactor system with internal aeration. Serum-supplemented and serum-free media comparison for MAb production. , 1994, Journal of immunological methods.

[22]  S. Jones,et al.  Increased monoclonal antibody ascites production in mice primed with Freund's incomplete adjuvant. , 1990, Journal of immunological methods.

[23]  C. Wraight,et al.  The effect of pristane on ascites tumor formation and monoclonal antibody production. , 1986, Methods in enzymology.

[24]  B. Brodeur,et al.  High yield monoclonal antibody production in ascites. , 1986, JIM - Journal of Immunological Methods.

[25]  M. Wüthrich,et al.  A simple and inexpensive high density dialysis tubing cell culture system for the in vitro production of monoclonal antibodies in high concentration. , 1993, Journal of immunological methods.

[26]  D. Garwes,et al.  Comparison of ascites production for monoclonal antibodies in BALB/c and BALB/c-derived cross-bred mice , 1989, Journal of Immunological Methods.

[27]  John Hopkinson,et al.  Hollow Fiber Cell Culture Systems for Economical Cell-Product Manufacturing , 1985, Bio/Technology.

[28]  J. Brandt,et al.  Influence of priming and inoculation dose on the production of monoclonal antibodies in two age groups of BALB/c mice. , 1994, Hybridoma.

[29]  L. Trudel,et al.  Evaluation of hollow fiber bioreactors as an alternative to murine ascites production for small scale monoclonal antibody production. , 1996, Journal of immunological methods.

[30]  S. Reuveny,et al.  Equipment and procedures for production of monoclonal antibodies in culture. , 1989, Advances in biotechnological processes.