The isolation of the α-toxin of Clostridium welchii type A by zone electrophoresis in vertical columns

Abstract

Culture filtrates of C. welchii type A concentrated by high-pressure ultrafiltration, have been examined by zone electrophoresis and immunoelectrophoresis. Zone electrophoresis was carried out with ethanolysed cellulose as the stabilizing medium in columns of a novel design. The columns were internally as well as externally cooled; a description of their construction is given. With such an apparatus, the a-toxin of C. welchii type A was quantitatively separated from the hyaluronidase, proteolytic and 9 -toxin activities. The lecithinase, hemolytic and lethal activities were electrophoretically inseparable. Immunoelectrophoresis was carried out on the toxin preparations. At least 9 different antigens were found in the one crude preparation and at least 25 in another. The [theta]-toxin obtained from the former preparation by zone electrophoresis was shown to be free from the other 8 antigens but was of low specific activity.