A novel method for the production of Salmonella flagellar antigen. II. Further purification for the preparation of H antisera.

Abstract

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.