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Use of monoclonal antibodies to identify the distribution of A and M epitopes on smooth Brucella species.

Department of Microbiology, University of Hawaii, Honolulu 96822.

ABSTRACT

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.

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