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 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.
Yajima
William
W
Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Alberta, Canada T6G 2P5.
Rahman
Muhammad H
MH
Das
Dipankar
D
Suresh
Mavanur R
MR
Kav
Nat N V
NN
D000595:Amino Acid Sequence; D000818:Animals; D000908:Antibodies, Fungal / Q000235:genetics / Q000276:immunology*; D000918:Antibody Specificity; D001203:Ascomycota / Q000302:isolation & purification*; D020171:Botrytis / Q000302:isolation & purification; D029688:Brassica napus / Q000382:microbiology; D019281:Dimerization; D015870:Gene Expression; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D025282:Mycelium / Q000302:isolation & purification; D019151:Peptide Library; D010935:Plant Diseases / Q000382:microbiology*
D000908:Antibodies, Fungal; D007135:Immunoglobulin Variable Region; D019151:Peptide Library