Successful development of monoclonal antibodies to three carnation viruses, using an admixture of only partially purified virus preparations as immunogen, and their use in virus diagnosis.

摘要

One of the stated advantages of hybridoma-produced monoclonal antibodies (McAbs), when compared with conventionally produced polyclonal antibodies, is the ability to produce and select target-specific McAbs even when impure antigen or antigen mixtures are used as immunogens. This 'advantage' was tested in a project designed to generate McAbs to carnation necrotic fleck closterovirus (CNFV), carnation latent carlavirus (CLV) and/or carnation mottle carmovirus (CarMV) viruses. An admixture of partially purified extracts from singly- and doubly-infected carnation plants was used as immunogen and individually as ELISA screening antigens (along with similarly prepared extracts from healthy plants). Extracts contained 5-20 protein bands as determined bySDS-PAGE electrophoresis. Thirty-eight hybridoma clones were selected, 27 to CNFV, 6 to CLV and 5 to CarMV. Virus specificity was confirmed by ELISA and Western-blot analysis. Selected McAbs have been used routinely in ELISA and tissue-print immunoassays for the detection of these viruses in carnation.