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Improved detection of Beet necrotic yellow vein virus in aDAS ELISA by means of antibody single chain fragments (scFv) which wereselected to protease-stable epitopes from phage display libraries

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Summary.

 The detection of Beet necrotic yellow vein virus (BNYVV) instored sugar beets by means of monoclonal antibodies or antibody singlechain fragments (scFv) often poses problems, because the immunodominantC-terminal epitope of the viral coat protein is readily lost due toproteolysis. Clones which produce scFv specific for protease-stableBNYVV epitopes were selected from two naive phage display libraries.Fusion proteins of the scFv with a human IgG kappa chain (expressed fromthe newly designed vector pCL) or with alkaline phosphatase,respectively, allow the ELISA detection of BNYVV even in stored sugarbeets with a sensitivity which was comparable or often higher than thatachieved with polyclonal antibodies.

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Authors and Affiliations

  1.  Biologische Bundesanstalt für Land -undForstwirtschaft, Institut für Pflanzenvirologie, Mikrobiologie undbiologische Sicherheit, Braunschweig, Germany, , , , , , DE

    K. Uhde & R. Koenig

  2.  Institut für angewandte Mikrobiologie, Universität für Bodenkultur, Wien, Austria, , , , , , AT

    R. J. Kerschbaumer, S. Hirschl & G. Himmler

  3.  Institut National de la Recherche Agronomique, Unite de Recherche Vigne et Vin, Equipe Vection, Colmar, France, , , , , , FR

    O. Lemaire

  4.  Central Science Laboratory, Sand Hutton, U.K., , , , , , GB

    N. Boonham

  5.  Cambridge Antibody Technology, U.K., , , , , , GB

    W. Roake

Authors
  1. K. Uhde
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  2. R. J. Kerschbaumer
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  3. R. Koenig
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  4. S. Hirschl
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  5. O. Lemaire
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  6. N. Boonham
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  7. W. Roake
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  8. G. Himmler
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Received July 26, 1999/Accepted August 13, 1999

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Uhde, K., Kerschbaumer, R., Koenig , R. et al. Improved detection of Beet necrotic yellow vein virus in aDAS ELISA by means of antibody single chain fragments (scFv) which wereselected to protease-stable epitopes from phage display libraries. Arch. Virol. 145, 179–185 (2000). https://doi.org/10.1007/s007050050015

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  • DOI: https://doi.org/10.1007/s007050050015

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