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Establishment of stable mouse/human-human hybrid cell lines producing large amounts of anti-tetanus human monoclonal antibodies with high neutralizing activity

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Abstract

To establish stable hybrid cell lines producing human anti-tetanus antibody with high toxinneutralizing activity, peripheral lymphocytes from humans hyperimmunized with tetanus toxoid were, after in vitro antigen stimulation, fused with a mouse/human heteromyeloma or human lymphoblastoid cell line and cloned. Unlike the IgM secretors (six clones), the IgG secretors we obtained (six clones) produced anti-tetanus human monoclonal antibodies with high neutralizing activity (the highest one, cell line G2, 4.3 IU/100 μg IgG).

Appropriate combinations of three or four kinds of monoclonal antibodies of the IgG type resulted in markedly increased neutralizing activity comparable with that of anti-tetanus human polyclonal immunoglobulin preparations currently used clinically on the basis of toxin-specific IgG content. Five of these cell lines produced 10≈20 pg of antibody per ml for more than 3 months. The cell line G2 produced 6 mg of antibody per day in serum-free medium in a 500-ml bioreactor in perfusion culture and 13–104 mg in a nude mouse. These cell lines satisfied, for the first time, the minimal requirements for applying human monoclonal antibodies to clinical use.

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Authors and Affiliations

  1. Morinaga Institute of Biological Science, 2-1-1 Shimosueyoshi, Tsurumi-ku, 230, Yokohama, Kanagawa, Japan

    M. Kamei & S. Hashizume

  2. Department of Tuberculosis Research, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, 565, Osaka, Japan

    N. Sugimoto, K. Ozutsumi & M. Matsuda

Authors
  1. M. Kamei
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  2. S. Hashizume
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  3. N. Sugimoto
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  4. K. Ozutsumi
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  5. M. Matsuda
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Kamei, M., Hashizume, S., Sugimoto, N. et al. Establishment of stable mouse/human-human hybrid cell lines producing large amounts of anti-tetanus human monoclonal antibodies with high neutralizing activity. Eur J Epidemiol 6, 386–397 (1990). https://doi.org/10.1007/BF00151713

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