Use of monoclonal antibodies to identify the distribution of A and M epitopes on smooth Brucella species.
The smooth types of Brucella species express two lipopolysaccharides (LPSs) (A and M) which are antigenically distinct. Their existence as cross-reactive antigenic complexes makes definitive serology difficult. Murine hybridomas were produced and selected for their ability to produce monoclonal antibodies to the specific A- or M-LPS epitopes. The specificity of the monoclonal antibodies was determined by microplate enzyme-linked immunoassay, binding inhibition assay, microplate agglutination, and dot blot assay. Monoclonal antibody 12AE6 was specific for an epitope on the A LPS of Brucella spp., which was also expressed on Yersinia enterocolitica O:9. A unique epitope of M LPS was detected by monoclonal antibody 33.1.5. The two monoclonal antibodies did not exhibit cross-reactions when assayed with whole Brucella cells or purified M- and A-LPS preparations. Cross-reactive serology with polyvalent sera can be attributed to the presence of common antigenic determinants on both molecules. The use of A- and M-LPS-specific monoclonal antibodies has the potential to replace the more laborious methods involved in the production of monospecific sera.
Douglas
J T
JT
Department of Microbiology, University of Hawaii, Honolulu 96822.
Palmer
D A
DA
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000942:Antigens, Bacterial / Q000032:analysis*; D001667:Binding, Competitive; D002002:Brucella / Q000276:immunology*; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D006825:Hybridomas; D007118:Immunoassay; D008070:Lipopolysaccharides / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D000939:Epitopes; D008070:Lipopolysaccharides