Summary
Mouse human-human heterohybridomas secreting human monoclonal antibodies (MoAb) against tetanus toxoid and hepatitis B virus surface antigen were effectively cultivated in a medium containing a serum substitute called GFS, a 55% to 70% ammonium sulphate fraction of serum from adult cattle. A perfusion culture system using a jar fermentor equipped with a cell sedimentation column with a double jacket was developed and applied to produce human MoAb. In this fermentor, maximum cell density of a heterohybridoma reached 1.2×107 cells/ml and MoAb was continuously accumulated at a constant rate for at least 40 days; this led to the production of more than one gram of human MoAb using a culture vessel with a 1-1 working volume.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Feder J, Tolbert WR (1983) The large-scale cultivation of mammalian cells. Scientific American 248:36–43
Glassy MC, Ferrone S (1982) Differential segregation patterns of human chromosomes in somatic cell hybrids constructed with human B-lymphocytes and human melanoma cells. Cancer Res 42:3971–3973
Himmelfarb P, Thayer PS, Martin HE (1969) Spin filter culture: The propagation of mammalian cells in suspension. Science 164:555–557
Ichimori Y, Sasano K, Itoh H, Hitotsumachi S, Kimura Y, Kaneko K, Kida M, Tsukamoto K (1985) Establishment of hybridomas secreting human monoclonal antibodies against tetanus toxin and hepatitis B virus surface antigen. Biochem Biophys Res Commun 129:26–33
Köhler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature (London) 256:495–497
Murakami H, Masui H, Sato GH, Sueoka N, Chow TP, Kano-Sueoka T (1982) Growth of hybridoma cells in serum-free medium: Ethanolamine is an essential component. Proc Natl Acad Sci USA 79:1158–1162
Nowinski R, Berglund C, Jane J, Lostrom M, Bernstein I, Young W, Hakomori S, Hill L, Cooney M (1980) Human monoclonal antibody against forssman antigen. Science 210:537–539
Olsson L, Kaplan HS (1980) Human-human hybridomas producing monoclonal antibodies of predefined antigenic specificity. Proc Natl Acad Sci USA 77:5429–5431
Ruddle F (1973) Linkage analysis in man by somatic cell genetics. Nature (London) 242:165–169
Sasai S, Umemoto S, Fujimoto T (1985) Establishment of a medium for mass culture of mammalian cells. Abst. 3rd Internation Cell Culture Congress (Sendai) p 46
Sato S, Kawamura K, Fujiyoshi N (1983) Mass propagation of animal cells on middle scale. Abst. 1st Symp. on Basic Technologies for Future Industries — Biotechnology — (Tokyo) p 119–128
Steinitz M, Klein G, Koskimies S, Makel O (1977) EB virus-induced B lymphocyte cell lines producing specific antibody. Nature 269:420–422
Teng NNH, Lam KS, Riesa RC, Kaplan HS (1983) Construction and testing of mouse-human heteromyelomas for human monoclonal antibody production. Proc Natl Acad Sci USA 80:7308–7312
Tolbert W, Feder J, Kimes RC (1981) Large-scale rotating filter perfusion system for high-density growth of mammalian suspension cultures. In Vitro 17:885–890
Truitt KE, Larrick JW, Raubitschek AA, Buck DW, Jacobson SW (1984) Production of human monoclonal antibody in mouse ascites. Hybridoma 3:195–199
Weiss MC, Green H (1967) Human-mouse hybrid cell lines containing partial complements of human chromosomes and functioning human genes. Proc Natl Acad Sci USA 58:1104–1111
Yelton DE, Diamond BA, Kwan SP, Scharff MD (1978) Fusion of mouse myeloma and spleen cells. Cur Top Microbiol Immunol 81:1–7
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kitano, K., Shintani, Y., Ichimori, Y. et al. Production of human monoclonal antibodies by heterohybridomas. Appl Microbiol Biotechnol 24, 282–286 (1986). https://doi.org/10.1007/BF00257050
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00257050