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Production of Monoclonal Antibodies Using a Secretion Capture Report Web

Bio/Technology volume 13, pages 787–790 (1995)Cite this article

Abstract

We describe a novel method for the production of monoclonal antibodies using a secretion capture report web (SCRW). Following HAT selection in bulk culture, individual hybridomas are encapsulated in biotinylated agarose drops. Antibody secreted by the hybridoma is captured within the agarose drop using an avidin bridge and biotinylated anti-mouse immunoglobulin. The secreted antibody is detected by a fluorescent reporter which can be either a second anti-mouse antibody or an antigen. The binding of the reporter can be quantitated and the desired hybridoma directly cloned by flow cytometry. Multiparameter (i.e., two-color) reporter analysis can also be used to selectively enrich and clone rare hybridomas secreting antibodies directed to unique epitopes. The method allows the characterization of thousands of clones per second and the isolation of hundreds of clones per day.

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Authors and Affiliations

  1. Department of Leukocyte Biology, Syntex Research, 3401 Hillview Ave., Palo Alto, California, 94304

    John S. Kenney & Marie-Hélène Ancel

  2. Department of Analytical and Structural Biology, Syntex Research, 3401 Hillview Ave., Palo Alto, California, 94304

    Forest Gray & John F. Dunne

  3. Department of Biophysics, UA491 CNRS, Université Louis Pasteur, Strasbourg, France

    John S. Kenney & Marie-Hélène Ancel

Authors
  1. John S. Kenney
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  2. Forest Gray
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  3. Marie-Hélène Ancel
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  4. John F. Dunne
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Correspondence to John S. Kenney.

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Kenney, J., Gray, F., Ancel, MH. et al. Production of Monoclonal Antibodies Using a Secretion Capture Report Web. Nat Biotechnol 13, 787–790 (1995). https://doi.org/10.1038/nbt0895-787

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