A novel method for the production of Salmonella flagellar antigen. II. Further purification for the preparation of H antisera.
A method for a simple preparation of Salmonella flagellar antigen is described. The antigen is sufficiently pure to elicit high titered H antibodies of 12,800-51,200 and O titers of less than 50. Highly motile Salmonella test strains are grown on 0.8% swarm agar and harvested with 0.05 n HCl which solubilizes the flagella. The suspension with a pH of 1.5 is kept at 4 degrees C. over night and then centrifuged at 49,000 g for 60 min. The supernatant is neutralized and precipitated with ammonium sulfate at 2/3 saturation. The resulting polymeric flagellin is submitted to a zone electrophoresis on Pevikon. Strips are cut from the "cake" and eluted. The H antigen is found on the anodic side, the O antigen remains near the trough or migrates slightly cathodically. Form 20 plates enough flagellin is collected fro the immunization of 50-100 rabbits. The Latex test proved to be especially suited for the checking of the H antigen.
D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D000942:Antigens, Bacterial / Q000302:isolation & purification*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D005407:Flagella / Q000276:immunology*; D005408:Flagellin / Q000276:immunology / Q000302:isolation & purification; D006385:Hemagglutination Inhibition Tests; D007122:Immunoelectrophoresis; D007841:Latex Fixation Tests; D011817:Rabbits; D012475:Salmonella / Q000276:immunology*; D013045:Species Specificity
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D005408:Flagellin