An antibody adsorption technique facilitates antigen selection for development of serotype-specific monoclonal antibodies to Yersinia enterocolitica.
A simple procedure is described to aid in the selection of antigens from gram-negative organisms that are most likely to exhibit serotype-specific epitopes when used as immunogens in the mouse. Several such proteins of Yersinia enterocolitica serotype O:8 were identified by immunoblotting through their binding to polyclonal antibodies that remained in immune serum after the serum had been adsorbed with common antigens prepared from a variety of closely related serotypes and Gram-negative organisms. The selected proteins were purified electrophoretically and used to immunize mice to obtain immune spleen cells for cell fusions. Serotype-specific monoclonal antibodies (MAbs) were successfully developed to a 36-kDa outer membrane protein resembling the porin OmpF. Additional MAbs also were produced that cross-reacted with other serotypes. Only cross-reactive epitopes were detected on a 34-kDa protein. The MAbs were used to show that the relative expression of the 36-kDa protein varied severalfold according to the medium used to grow the cells. These MAbs are promising agents for serological identification of Y. enterocolitica serotype O:8. The procedure should be useful in many situations where extensive cross-reactivities impede generation of highly specific MAbs.
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Department of Bioscience and Biotechnology, Drexel University, Philadelphia, PA 19104.
Magee
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D000327:Adsorption; D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis* / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000942:Antigens, Bacterial / Q000276:immunology*; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis / Q000276:immunology; D003429:Cross Reactions; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D012703:Serotyping; D015008:Yersinia enterocolitica / Q000145:classification / Q000276:immunology*
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins