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BALB/c mice were immunized with Corynebacterium sepedonicum, and spleen cells from the immunized animals were fused with cells of the mouse myeloma line P3-X63-Ag8.653. Several hybridoma cell cultures were selected for further study. Monoclonal CS-B-5 was specific for C. sepedonicum and did not react significantly with other closely related phytopathogenic corynebacteria in an enzyme-linked immunosorbent assay (ELISA). As few as 10(3) organisms could be detected. This approach should prove useful for developing improved diagnostic procedures for a number of bacterial plant pathogens.

We describe a novel approach of producing monoclonal antibodies (MABs) to one specific protein of a virus or other agent consisting of several proteins, without the use of purified antigen in either the immunization or screening phase of the procedure. This method has general application in the production of MABs when the antigen cannot be obtained in a pure form, but the gene is available. We illustrate this application by producing MAB specific to the nucleocapsid protein (N) of vesicular stomatitis virus serotype Indiana (VSV-IN) from BALB/c mice immunized with an infectious vaccinia virus recombinant vector (v38) that expresses the N gene of VSV-IN. This novel method of immunization obviates the need for initial purification of the protein antigen and injection of adjuvants with the isolated protein as is done in traditional MAB production.