Virus strain discrimination using recombinant antibodies

Article type: Research Article

Authors: N. Boonham, | I. Barker,

Affiliations: Central Science Laboratory, Sand Hutton, York YO41 1LZ, UK. Tel.: +44 1904 462000; Fax: +44 1904 462111; E-mail: [email protected].

Abstract: 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 {\it Potato virus Y}, ordinary strain (PVY^{\rm O}). The scFv selected was specific for PVY and detected 7 out of 9 isolates of PVY^{\rm O} whilst it did not detect 15 isolates from the closely related necrotic strains PVY^{\rm N}and PVY^{\rm NTN}. 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.

Journal: Disease Markers, vol. 16, no. 1-2, pp. 95-97, 2000