Open AccessCharacterization of immunogenic p230 as the toxin A of Clostridium difficile
Author(s) -
Rautenberg P.,
Stender F.
Publication year - 1986
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1986.tb01756.x
Subject(s) - enterotoxin , toxin , clostridium difficile toxin a , antiserum , clostridium difficile , microbiology and biotechnology , clostridiaceae , polyacrylamide gel electrophoresis , molecular mass , clostridium , gel electrophoresis , chemistry , biology , biochemistry , antibody , bacteria , escherichia coli , enzyme , immunology , genetics , gene , antibiotics
For the identification of toxin A of Clostridium difficile , a 2‐dimensional gel system was used. In its first dimension, samples were separated in the absence of reducing and dissociating agents, conditions which maintained the activity of the enterotoxin. This was followed by reduction and dissociation in the second dimension where a 230 kDa polypeptide was electroeluted. Rabbits were immunized with polyacrylamide gel slices containing entrapped native toxin A and the denatured 230 kDa protein. As revealed by immunoblotting, neutralizing antisera derived from native protein samples recognized the native toxin, the denatured 230 kDa protein and another polypeptide of about M r 35 000. Using both types of antisera as probes the pI of the enterotoxin was about 5.9. Preliminary evidence suggests that the enterotoxin is a multimeric protein of 230 kDa and 35 kDa subunits.