Articles
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
Article number
568_26
Pages
177 – 183
Language
English
Abstract
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 by SDS-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.
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 by SDS-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.
Authors
R. Jordan, M.A. Guaragna
Keywords
ELISA, carnation necrotic fleck closterovirus, carnation latent carlavirus
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