Monoclonal antibodies for detection, serological characterization and immunopurification of grapevine fleck virus

Ten stable hybridoma cell lines secreting monoclonal antibodies to grapevine fleck virus (GFkV) were selected after fusing spleen cells of immunized Balb/C mice with mouse myeloma cells (SP2/0-Ag 14). All MAbs reacted positively in ELISA with leaf extracts from fifty GFkV-infected grapevines from various geographical origins. MAb 2B5 was used for routine detection of GFkV and appeared to be more sensitive than polyclonal antibodies. The first attempt to purify GFkV by immunoaffinity chromatography using MAb 2B5 led to highly purified coat protein. This procedure encompassed fewer steps and allowed the use of tissues other than rootlets for satisfactory purification.