VIEW ARTICLE

Techniques

Production of Monoclonal Antibodies to Corynebacterium sepedonicum. S. H. De Boer, Agriculture Canada Research Station, Vancouver, B.C., Canada, V6T 1X2; A. Wieczorek, Agriculture Canada Research Station, Vancouver, B.C., Canada, V6T 1X2. Phytopathology 74:1431-1434. Accepted for publication 25 June 1984. Copyright 1984 The American Phytopathological Society. DOI: 10.1094/Phyto-74-1431.

Five hybridoma cell lines, VRS-H-004-CS3-1 to VRS-H-008-CS3-5, secreting monoclonal antibodies McAb 1 to McAb 5, respectively, to Corynebacterium sepedonicum were produced by fusing splenocytes from immunized BALB/c mice with P3/NS1/1-AG4-1 (NS-1) myeloma cells. Both whole cells and a lithium chloride cell extract were used as immunogen and as antigen in enzyme-linked immunoassays for screening hybridomas. Monoclonal antibodies produced by all five hybridomas reacted in indirect immunofluorescence with 19 strains of C. sepedonicum from 18 different geographic areas. McAb 3 and 4 reacted with strains of C. fascians, C. insidiosum, C. michiganense pv. iranicum, and C. poinsettiae. McAb 1 reacted weakly with C. fascians and C. insidiosum. McAb 2 and 5 did not react with any of the ten other plant pathogenic corynebacteria species tested. All the monoclonal antibody preparations except McAb 5 reacted with one or more unidentified nonpathogenic bacteria isolated from potato.