title: Improved detection of Beet necrotic yellow vein virus in a DAS ELISA by means of antibody single chain fragments (scFv) which were selected to protease-stable epitopes from phage display libraries.
abstract: The detection of Beet necrotic yellow vein virus (BNYVV) in stored sugar beets by means of monoclonal antibodies or antibody single chain fragments (scFv) often poses problems, because the immunodominant C-terminal epitope of the viral coat protein is readily lost due to proteolysis. Clones which produce scFv specific for protease-stable BNYVV epitopes were selected from two naive phage display libraries. Fusion proteins of the scFv with a human IgG kappa chain (expressed from the newly designed vector pCL) or with alkaline phosphatase,respectively, allow the ELISA detection of BNYVV even in stored sugar beets with a sensitivity which was comparable or often higher than that achieved with polyclonal antibodies.
authors.affiliation: Biologische Bundesanstalt für Land -undForstwirtschaft, Institut für Pflanzenvirologie, Mikrobiologie undbiologische Sicherheit, Braunschweig, Germany.
authors.forename: K
authors.initials: K
authors.lastname: Uhde
authors.forename: R J
authors.initials: RJ
authors.lastname: Kerschbaumer
authors.forename: R
authors.initials: R
authors.lastname: Koenig
authors.forename: S
authors.initials: S
authors.lastname: Hirschl
authors.forename: O
authors.initials: O
authors.lastname: Lemaire
authors.forename: N
authors.initials: N
authors.lastname: Boonham
authors.forename: W
authors.initials: W
authors.lastname: Roake
authors.forename: G
authors.initials: G
authors.lastname: Himmler
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D001435:Bacteriophages / Q000235:genetics; D019663:Chenopodiaceae / Q000821:virology*; D010450:Endopeptidases / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D019151:Peptide Library; D010935:Plant Diseases / Q000821:virology; D010942:Plant Viruses / Q000276:immunology / Q000302:isolation & purification*; D012328:RNA Viruses / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D007128:Immunoglobulin Fragments; D019151:Peptide Library; D010450:Endopeptidases