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Highly efficient targeting and accumulation of a Fab fragment within the secretory pathway and apoplast of Arabidopsis thaliana

  • Peeters, Koen
  • De Wilde, Chris
  • Depicker, Anna
Publication Date
Jan 01, 2001
Ghent University Institutional Archive
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To further improve antibody production in plants, constructs were designed to minimize transgene silencing and to retain a F-ab fragment within the secretory pathway of transgenic Arabidopsis thaliana plants. The levels of antibody accumulation suggest that placing the sequences that encode Fd and light chain under the control of nonidentical 3' regions reduces susceptibility to posttranscriptional gene silencing compared with when the individual polypeptide-encoding sequences are placed under the control of identical 3' regions. High levels of accumulation (up to 6% of total soluble protein) were found for both secreted and intracellularly targeted antibody fragments. Immunofluorescence microscopic analysis showed that F-ab fragments devoid of any additional C-terminal sequence were efficiently secreted, whereas retention of F-ab fragments within the endomembrane system of the secretory pathway was achieved by C-terminal fusion of the DIKDEL sequence to the antibody tight chain. Furthermore, analysis by immunoprecipitation and ELISA showed that intracellular retention of antibody fragments did not affect antigen-binding activity, and more than 80% of the isolated antibody fragments were found to bind antigen. Taken together, our results provide improvements to the technology of recombinant antibody production in transgenic plants.

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