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Synthesis and self-assembly of a functional monoclonal antibody in transgenic Nicotiana tabacum

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Abstract

Immunoglobulin light and heavy chains are synthesized in mammalian cells as precursors containing a signal peptide. Processing and assembling result in formation of active antibodies. Chimeric genes have been made containing the coding sequence of the barley α-amylase signal peptide which has been fused to cDNAs coding for either the mature light or the mature heavy chain of a monoclonal antibody. A plasmid was constructed linking both chimeric genes under the control of plant active promoters in an expression cassette. This DNA fragment was stably integrated into the genome of Nicotiana tabacum by Agrobacterium tumefaciens mediated gene transfer. Synthesis of light and heavy chains and assembly to antibodies was detected in transgenic tobacco tissue using specific secondary antibodies. By electron microscopic immunogold labeling, the presence of assembled antibody could be detected within the endoplasmic reticulum. Affinity chromatography indicated biological activity of the assembled immunoglobulin produced in plant cells. Unexpectedly, a significant amount of assembled antibodies was found within chloroplasts.

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Author notes

  1. Klaus Düring

    Present address: Center for Applied Plant Molecular Biology, University of Hamburg, Ohnhorststr. 18, D-2000, Hamburg 52, FRG

Authors and Affiliations

  1. Max Planck Institute for Plant Breeding, D-5000, Köln, FRG

    Klaus Düring, Fritz Kreuzaler & Jeff Schell

  2. Institute for Biology III, RWTH Aachen, D-5100, Aachen, FRG

    Sigrun Hippe

  3. Institute for Biology I, RWTH Aachen, D-5100, Aachen, FRG

    Fritz Kreuzaler

Authors
  1. Klaus Düring
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  2. Sigrun Hippe
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  3. Fritz Kreuzaler
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  4. Jeff Schell
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Düring, K., Hippe, S., Kreuzaler, F. et al. Synthesis and self-assembly of a functional monoclonal antibody in transgenic Nicotiana tabacum . Plant Mol Biol 15, 281–293 (1990). https://doi.org/10.1007/BF00036914

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  • DOI: https://doi.org/10.1007/BF00036914

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