Cucumber mosaic cucumovirus antibodies from a synthetic phage display library.
Antibody fragments (scFv) that bind specifically to particles of cucumber mosaic cucumovirus (CMV) were obtained from a library which encodes a diverse array of synthetic antibody fragments, each displayed on the surface of filamentous bacteriophage. After four rounds of selection and enrichment, several clones were obtained which produced scFv that bound specifically to purified particles of CMV in ELISA. BstNI digestion of phagemid DNA resulted in the same restriction pattern for all clones. The nucleotide sequences of three of the clones showed that they belonged to the human VH1 family and that they had a complementarity determining region loop of 7 amino acids. Phage-displayed antibodies and soluble scFv secreted by these clones reacted with particles of CMV in sap from infected plants in ELISA. In immunoblotting tests, soluble scFv preparations reacted with SDS-denatured coat protein extracted from purified preparations of CMV isolates belonging to either subgroup I or II and also with protein extracted by SDS treatment of seeds harvested from naturally infected lupin plants. The results demonstrate the feasibility, and potential applicability, of recombinant antibody methods in plant pathology.
Virology Department, Scottish Crop Research Institute, Invergowrie, Dundee, United Kingdom.
A
A
Ziegler
L
L
Torrance
S M
SM
Macintosh
G H
GH
Cowan
M A
MA
Mayo
D000595:Amino Acid Sequence; D000914:Antibodies, Viral / Q000276:immunology*; D001435:Bacteriophages / Q000235:genetics / Q000276:immunology; D001483:Base Sequence; D018553:Cucumis sativus / Q000821:virology; D017799:Cucumovirus / Q000235:genetics / Q000276:immunology*; D004279:DNA, Viral; D015723:Gene Library; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D014771:Virion / Q000276:immunology
D000914:Antibodies, Viral; D004279:DNA, Viral; D007128:Immunoglobulin Fragments; D011994:Recombinant Proteins