PRODUCTION AND CHARACTERIZATION OF MONOCLONAL ANTIBODIES TO CLOSTRIDIUM PERFRINGENS ENTEROTOXIN DERIVED FROM DIFFERENT ANIMAL SPECIES

The enterotoxins (ETs) of Clostridium perfringens isolated from enterotoxemic or diarrheic alpacas, pigs, calves, dogs, and horses were obtained from sporulated cell extracts. The ETs from alpaca, pig, and calf isolates were chromatographed on Sephadex G-lOO. Monoclonal antibodies (MAbs) against ETs derived from alpaca, pig, and calf C. perfringens isolates were produced. The MAbs were used in neutralization of cytotoxicity and immunoblot assays to study cross-reactivity among all five ETs. Using neutralization of cytotoxicity technique, each MAb exhibited neutralization against the cytopathic effect of all five ETs on Vero cells. The neutralization ratio in this study was I ng ET: 50 ng MAb. On immunoblots, the anti-alpaca ET and anti-pig ET cross-reacted with partially purified ETs from pig and alpaca respectively. In addition, anti-alpaca ET and anti-pig ET crossreacted with partially purified ETs from calf and horse, but not ET from dog isolates. The anti-calf ET cross-reacted with alpaca, calf, dog, and pig but not with ET from the horse isolate. The immunoblots also indicated that the MAbs recognized aggregated ET subunits. Our data indicated that C. perfringens ETs derived from different animal species are very similar.