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

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 pr...

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Bibliographic Details
Published in:Archives of virology 2000-01, Vol.145 (1), p.179-185
Main Authors: UHDE, K, KERSCHBAUMER, R. J, KOENIG, R, HIRSCHL, S, LEMAIRE, O, BOONHAM, N, ROAKE, W, HIMMLER, G
Format: Article
Language:English
Subjects:
Antibodies, Monoclonal - immunology
Bacteriophages - genetics
Beet necrotic yellow vein virus
Biological and medical sciences
Chenopodiaceae - virology
Endopeptidases - metabolism
Enzyme-Linked Immunosorbent Assay - methods
Epitopes
Fundamental and applied biological sciences. Psychology
Generalities. Techniques. Transmission, epidemiology, ecology. Antiviral substances, control
Humans
Immunoglobulin Fragments - genetics
Immunoglobulin Fragments - immunology
Life Sciences
Microbiology
Microbiology and Parasitology
Molecular Sequence Data
Peptide Library
Phytopathology. Animal pests. Plant and forest protection
Plant Diseases - virology
Plant Viruses - immunology
Plant Viruses - isolation & purification
Plant viruses and viroids
RNA Viruses - immunology
RNA Viruses - isolation & purification
single-chain variable fragment antibody
Techniques used in virology
Virology
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Summary: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.
ISSN:0304-8608
1432-8798
DOI:10.1007/s007050050015