Characterization of Shigella flexneri-specific murine monoclonal antibodies by chemically defined glycoconjugates.
Chemically defined glycoconjugates are demonstrated to have considerable potential for selecting hybridoma antibodies directed toward O-antigenic determinants, especially when used in combination with a panel of well-characterized LPS molecules. Monoclonal antibodies specific for the Shigella flexneri O-antigens of serogroup 5b, variants X and Y, were generated after immunization of BALB/c mice with killed bacterial cells, and active hybrids were selected on the basis of ELISA performed with the purified serotype-specific LPS antigen. Subsequent screening with a variety of glycoconjugates, derived from synthetic oligosaccharides and larger structures obtained by phage Sf6/endo-rhamnosidase hydrolysis of purified LPS established a detailed profile of binding characteristics for Shigella flexneri variant Y-specific antibodies. Together with the results of precipitin analysis and heavy chain isotyping experiments, a limited number of antibodies were selected as candidates for detailed studies of the antibody combining site.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D000942:Antigens, Bacterial / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D051379:Mice; D011135:Polysaccharides, Bacterial / Q000138:chemical synthesis / Q000276:immunology*; D012703:Serotyping; D012763:Shigella flexneri / Q000145:classification / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D011135:Polysaccharides, Bacterial