Detection of Sclerotinia sclerotiorum Using a Monomeric and Dimeric Single-Chain Fragment Variable (scFv) Antibody

Sclerotinia sclerotiorum (Lib.) de Bary is a phytopathogenic fungus capable of causing significant yield losses in numerous crops, including canola, in which the fungus causes sclerotinia stem rot. Immunological detection methods to rapidly determine the presence of S. sclerotiorum on plants may pro...

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Bibliographic Details
Published in:Journal of agricultural and food chemistry 2008-10, Vol.56 (20), p.9455-9463
Main Authors: Yajima, William, Rahman, Muhammad H, Das, Dipankar, Suresh, Mavanur R, Kav, Nat N. V
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
antibodies
Antibodies, Fungal - genetics
Antibodies, Fungal - immunology
Antibody Specificity
Ascomycota - isolation & purification
Biological and medical sciences
Botrytis - isolation & purification
Botrytis cinerea
Brassica napus - microbiology
Brassica napus var. napus
Canola
Chemical Aspects of Biotechnology/Molecular Biology
diagnostic techniques
Dimerization
dose response
flowers
Food industries
Food microbiology
Fundamental and applied biological sciences. Psychology
Gene Expression
General aspects
immunoassays
Immunoglobulin Variable Region - genetics
Immunoglobulin Variable Region - immunology
immunologic techniques
Methods of analysis, processing and quality control, regulation, standards
Mice
Mice, Inbred BALB C
Molecular Sequence Data
Mycelium - isolation & purification
Peptide Library
phage display
phytopathogen
Plant Diseases - microbiology
plant pathogenic fungi
rapid methods
scFv
Sclerotinia sclerotiorum
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Summary:Sclerotinia sclerotiorum (Lib.) de Bary is a phytopathogenic fungus capable of causing significant yield losses in numerous crops, including canola, in which the fungus causes sclerotinia stem rot. Immunological detection methods to rapidly determine the presence of S. sclerotiorum on plants may provide growers with a viable diagnostic tool to aid with fungicide use decisions. This paper discusses the generation of a monomeric and dimeric single-chain, variable fragment (scFv) antibody with affinity for S. sclerotiorum using phage display technology. The bacterially expressed and purified scFv is shown to bind S. sclerotiorum with some cross-reactivity with the closely related phytopathogen Botrytis cinerea (Pers.:Fr.). The dimeric scFv displayed improved binding to the fungus as compared to the monomer and could detect the presence of mycelia in inoculated canola petals. To the authors’ knowledge, this is the first report of a scFv dimer with affinity for S. sclerotiorum that has the potential for use in the development of a new diagnostic test.
ISSN:0021-8561
1520-5118
DOI:10.1021/jf801768g