Use of monoclonal antibodies to detect a phytotoxic glycopeptide produced by Ophiostoma ulmi, the Dutch elm disease pathogen

Abstract

Two hybridomas that secrete antibodies specific for a phytotoxic glycopeptide from Ophiostoma ulmi (Buism.) Nannf. were produced by fusing spleen cells of mice immunized with the purified toxin and the Sp2-0/Ag14 mouse myeloma cell line. Specificity of these antibodies was first demonstrated by an enzyme-linked immunosorbent assay (ELISA), then by immunoblotting on nitrocellulose membrane after sodium dodecyl sulfate – polyacrylamide gel electrophoresis of the glycopeptide. Both clones produced antibodies of IgM class as determined by immunodiffusion. These monoclonal antibodies were utilized to detect and localize the toxic glycopeptide in pathogen cells and infected host tissues by immunohistochemical and immunocytochemical techniques.