Abstract
Mouse-human heterohybridoma cell line, GF4/1.1, secretes a human anti-tetanus toxoid (anti-TT) antibody of the γ3 isotype. This line was used as a source of mRNA for the construction of cDNA libraries and the subsequent isolation of the sequences encoding the human heavy and light chains. Expression vectors containing the cDNAs were constructed and introduced into a non-Ig producing, murine hybridoma cell line, together with a dihy-drofolate reductase (dhfr) selectable marker gene. Transfected cells producing human anti-TT antibody were grown in medium with increasing concentrations of the selecting drug (methotrexate). The level of antibody secretion was tested in cells that had adapted to the higher drug concentration and was found to increase in parallel with increases in the steady-state level of Ig mRNA. This increased productivity was not associated with detectable levels of gene amplification. Stable clones were obtained that secrete levels of human antibody in either open suspension cultures or within microcapsules that compare favorably with those of murine hybridomas.
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References
James, K. and Bell, G.T. 1987. Human monoclonal antibody production. Current status and future prospects. J. Immunol. Methods. 100:5–40.
Boyd, J.E. and James, K. 1988. Human monoclonal antibodies: Their potential, problems and prospects. In: Advances in Biotechnological Processes. Vol. 11. Mizrahi, A. (Ed.). A. R. Liss, Inc., New York.
Larrick, J.W. and Bourla, J.M. 1986. The prospects for the therapeutic use of human monoclonal antibodies. J. Biol. Res. Mod. 5:379–393.
Morrison, S.L., Johnson, M.J., Herzenberg, L.A. and Oi, V.T. 1984. Chimeric human antibody molecules: mouse antigen-binding domains with human constant region domains. Proc. Natl. Acad. Sci. U.S.A. 81:6851–6855.
Jones, P.T., Dear, P.H., Foote, J., Neuberger, M.S. and Winter, G. 1986. Replacing the complementarity-determining regions in a human antibody with those from a mouse. Nature 321:522–525.
McCune, J.M., Namikawa, R., Kaneshima, H., Shultz, L.D., Lieberman, M. and Weissman, I.L. 1988. The SCID-hu mouse: murine model for the analysis of human hematolymphoid differentiation and function. Science 241:1632–1639.
Mosier, D.E., Gulizia, R.J., Baird, S.M. and Wilson, D.B. 1988. Transfer of a functional immune system to mice with severe combined immunodeficiency. Nature 335:256–259.
Gubler, U. and Hoffman, B.J. 1983. A simple and very effective method for generating cDNA libraries. Gene 25:263–269.
Huynh, T., Young, R. and Davis, R. 1984. Construction and screening of cDNA libraries in λgt10 and λgt11. In: DNA Cloning, Vol. 1: A practical approach. Glover, D. (Ed.). IRL Press, Oxford.
Gillies, S.D. 1986. Vector and method for achieving high level expression in eukaryotic cells. Western European Patent Application No. 8730 0658. 9.
Mulligan, R.C. and Berg, P. 1981. Selection for animal cells that express the E. coli gene coding for xanthine-guanine phosphoribosyl-transferase. Proc. Natl. Acad. Sci. U.S.A. 78:2072–2076.
Simonsen, C.C. and Levinson, A.D. 1983. Isolation and expression of an altered mouse dihydrofolate reductase cDNA. Proc. Natl. Acad. Sci. U.S.A. 80:2495–2499.
Galfre, G. and Milstem, C. 1981. Preparation of monoclonal antibodies: strategies and procedures. Meth. in Enzymol. 73:3–46.
Ochi, A., Hawley, R.G., Shulman, M.J., Traunecker, A., Kohler, G. and Hozumi, N. 1983. Functional immunoglobulin M production after transfection of cloned immunoglobulin heavy chains into lymphoid cells. Proc. Natl. Acad. Sci. U.S.A. 78:6351–6355.
Gillies, S.D., Morrison, S.L., Oi, V.T. and Tonegawa, S. 1983. A tissue-specific transcriptional enhancer element is located in the major intron of a rearranged immunoglobulin heavy-chain gene. Cell 33:717–728.
Rupp, R.G. and Geyer, S.D. 1984. Preparation of medium for large-scale hybridoma culture. J. Tissue Cult. Meth. 8:141–145.
Jarvis, A. and Grdina, T.A. 1983. Production of biologicals from microencapsulated living cells. BioTechniques 1:22–27.
Bell, G.T. 1987. Studies on the production of human monoclonal antibodies. PhD Thesis. University of Edinburg.
Jirik, F.R., Sorge, J., Fong, S., Heitzmann, J.G., Curd, J.G., Chen, P.P., Goldfien, R. and Carson, D.A. 1986. Cloning and sequence determination of a human rheumatoid factor light-chain gene. Proc. Natl. Acad. Sci. U.S.A. 83:2195–2199.
Schimke, R.T. 1988. Gene amplification in cultured cells. J. Biol. Chem. 263:5989–5992.
Rupp, R.G. 1986. Use of cellular microencapsulation in large-scale production of monoclonal antibodies. In: Large-Scale Mammalian Cell Culture. Tolbert, W.R. (Ed.). Academic Press, New York.
Brown, B.A., Davis, G.L., Salzgaber-Muller, J., Simon, P., Ho, M.-K., Shaw, P.S., Stone, B.A., Sands, H. and Moore, G.P. 1987. Tumor-specific genetically engineered murine/human chimeric monoclonal antibody. Canc. Res. 47:3577–3583.
Gillies, S.D. and Lo, K.-M. 1989. Expression of chimeric antibodies. In: Production of Therapeutic Biologicals. Jarvis, A. (Ed.). Marcel Dekker, Inc. New York.
Dorai, H. and Moore, G.P. 1987. The effect of DHFR-mediated gene amplification on the expression of transfected immunoglobulin genes. J. Immunol. 139:4232–4241.
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Gillies, S., Dorai, H., Wesolowski, J. et al. Expression of Human Anti-Tetanus Toxoid Antibody in Transfected Murine Myeloma Cells. Nat Biotechnol 7, 799–804 (1989). https://doi.org/10.1038/nbt0889-799
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DOI: https://doi.org/10.1038/nbt0889-799
