Expression of Human Anti-Tetanus Toxoid Antibody in Transfected Murine Myeloma Cells
Author: ["Stephen D. Gillies","Hamanti Dorai","John Wesolowski","Gerry Majeau","Delano Young","Jan Boyd","John Gardner","Keith James"]
Publication: Bio/Technology
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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.
Cite this article
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
