title: Characterization of six murine monoclonal antibodies specific for toxin B of Clostridium difficile.
abstract: Six murine hybridoma cell lines producing monoclonal antibodies (MAbs) specific for Toxin B of Clostridium difficile have been generated from toxin-immunized female RBF/DnJ mice. All six antibodies were reactive in Western blots with a > 200-kD protein in the supernatants of the toxigenic strain 10463 and were unreactive with similarly prepared material from the nontoxigenic strain 2037. Polyclonal antisera from rabbits immunized with Toxin B reacted on Western blots primarily with Toxin B, a 40-kD and a 55-kD band with a minor set of triplet bands at approximately 100 kD. None of the MAbs reacted in a direct EIA with purified Toxin A from C. difficile but two MAbs reacted weakly with a trypsin-sensitive band (> 200 kD) in Western blots of C. sordellii. Polyclonal antisera developed against Toxin B reacted strongly with supernatants from C. sordellii, C. bifermentans, and the nontoxigenic strain 2037. Toxin B-specific antisera was unreactive with supernatants from C. perfringens or purified Toxin A from C. difficile in direct EIA. Toxin B-specific MAbs linked to an affinity column were able to deplete bacterial supernatant of cytotoxigenic activity.
authors.affiliation: Cambridge Biotech Corporation, Worcester, MA 01605.
authors.forename: R T
authors.initials: RT
authors.lastname: Coughlin
authors.forename: M
authors.initials: M
authors.lastname: Annunziato
authors.forename: J
authors.initials: J
authors.lastname: Roberson
authors.forename: D J
authors.initials: DJ
authors.lastname: Marciani
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000942:Antigens, Bacterial / Q000276:immunology* / Q000302:isolation & purification; D001426:Bacterial Proteins; D001427:Bacterial Toxins / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western; D016360:Clostridioides difficile / Q000276:immunology*; D005260:Female; D006825:Hybridomas; D007124:Immunoenzyme Techniques; D051379:Mice; D008815:Mice, Inbred Strains; D011817:Rabbits
chemical_list: D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D001427:Bacterial Toxins; C031815:toxB protein, Clostridium difficile