Monoclonal antibodies against alpha toxin of Clostridium perfringens.
Ten distinct monoclonal antibodies (MAbs) against alpha toxin of Clostridium perfringens were produced by the fusion of SP2/O with spleen cells of mice immunized with alpha toxoid, and alpha toxin mixed with or without ethylenediamine-tetraacetate (EDTA). The antibody activity was evaluated by antigen-binding activity in an enzyme linked immunosorbent assay (ELISA), by phospholipase C (PLC)-neutralizing activity using both egg yolk lecithin and p-nitrophenylphosphoryl-choline (PNPPC) hydrolysis reactions and by anti-lethal activity in mice. Since the toxin-neutralizing activities of each MAb were not parallel, it has been suggested that the three biological activities may not be located in the same site in the toxin molecule. This report also describes the development of a simple purification of the toxin by affinity chromatography and a sensitive immunoassay for quantitation of the toxin using the monoclonal antibody.
Sato
H
H
Department of Applied Immunology, National Institute of Health, Tokyo, Japan.
Chiba
J
J
Sato
Y
Y
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001427:Bacterial Toxins / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D002135:Calcium-Binding Proteins; D002846:Chromatography, Affinity; D003016:Clostridium perfringens / Q000472:pathogenicity*; D051379:Mice; D008807:Mice, Inbred BALB C; D010738:Type C Phospholipases
D000911:Antibodies, Monoclonal; D001427:Bacterial Toxins; D002135:Calcium-Binding Proteins; D010738:Type C Phospholipases; C040649:alpha toxin, Clostridium perfringens