Virus strain discrimination using recombinant antibodies.
Most routine testing for plant viruses is currently carried out using monoclonal and polyclonal antibodies. Traditional methods of antibody production however can be time consuming and require the use of expensive cell culture facilities. Recombinant antibody technology however is starting to make an impact in this area, enabling the selection of antibody fragments in a few weeks compared with the many months associated with traditional methods and requires only basic microbiological facilities. Single chain Fv antibody fragments (scFv) have been selected from a synthetic phage-antibody library by affinity selection with purified Potato virus Y, ordinary strain (PVYO). The scFv selected was specific for PVY and detected 7 out of 9 isolates of PVYO whilst it did not detect 15 isolates from the closely related necrotic strains PVYN and PVYNTN. In ELISA the scFv could be used to detect virus at concentrations of 50 ng/ml in plant sap and was shown to have similar limits of detection as commercially available PVY monoclonal antibodies. These results highlight the potential of the technology for the selection of strain specific antibodies with an affinity and assay sensitivity similar to traditional monoclonal antibodies and their use in viral diagnostics.
Boonham
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Central Science Laboratory, Sand Hutton, York YO41 1LZ, UK. n.boonham@csl.gov.uk
Barker
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I
D000914:Antibodies, Viral / Q000235:genetics*; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D005803:Genes, Immunoglobulin; D007128:Immunoglobulin Fragments / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D017800:Potyvirus / Q000145:classification / Q000235:genetics; D011994:Recombinant Proteins; D014764:Viral Proteins / Q000032:analysis; D014780:Viruses / Q000145:classification* / Q000235:genetics*
D000914:Antibodies, Viral; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins; D014764:Viral Proteins