Monoclonal antibodies to Bacteroides fragilis lipopolysaccharide

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Abstract
Monoclonal antibodies (MoAbs) to the lipopolysaccharide (LPS) of B. fragilis were produced by immunizing mice before hybridization with bacterial outer membranes solubilized with Triton X-100. Nineteen stabile clones were established. They all produced antibodies that reacted more strongly with purified B. fragilis LPS than with crude sonicated antigen in an enzyme immunoassay. Four MoAbs were studied by immunoblotting and enzyme immunoassay inhibition. Immunoblotting confirmed that the target of the MoAbs was LPS. Marked and homogeneous staining occurrred in the immunoblotting with both purified LPS and outer membranes in the MW range 8000-27,000. In enzyme immunoassay inhibition, MoAbs reacted positively with 93-96% of B. fragilis strains, including prototype strains ATCC 23745 and NCTC 9343. Within the B. fragilis group, the MoAbs reacted positively with 2 of 5 B. ovatus strains and 2-6 of 9 B. thetaiotaomicron strains. No marked cross-reactivity with other bacteria was observed. These results confirm earlier findings that the B. fragilis LPS contains an immunodominant antigenic determinant common to almost all B. fragilis isolates.

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