Specificity of antibodies to the reconstituted crotoxin complex, from the venom of South American rattlesnake ( Crotalus durissus terrificus), using enzyme-linked immunosorbent assay (ELISA) and double immunodiffusion

Abstract

Some differences between the reaction of antiserum to reconstituted crotoxin complex, to ‘native’ crotoxin and to whole Crotalus durissus terrificus (South American rattlesnake) venom were detected by enzyme-linked immunosorbent assay (ELISA) and by immunodiffusion. ELISA showed the presence of antibodies to the components of the crotoxin complex, phospholipase A 2 and crotapotin. When the antiserum was tested against North American rattlesnake venoms some species gave a positive reaction ( C. horridus atricaudatus and C. basiliscus), whilst the venoms of other species were virtually negative ( C. durissus totonacus, C. horridus horridus, C. viridis viridis and C. atrox). Confirmation of antigenic similarities between crotoxin and Mojave toxin (from the venom of C. scutulatus scutulatus) was obtained with ELISA and some implications of these species differences in antigen-antibody reaction are discussed. No paraspecificity was observed with heterologous snake venoms from other Crotalidae, Elapidae, Hydrophiidae or Viperidae. ELISA also showed a lack of cross-reactivity of the antiserum to heterologous purified phospholipases A 2 from Enhydrina schistosa, Naja nigricollis or Apis mellifera venoms or from porcine pancreas. The antiserum reacted to the homologous phospholipase A 2 inactivated with p-bromophenacyl bromide as well as to the detoxified crotoxin complex reconstituted with this modified phospholipase A 2. This may be useful in the raising of high titres of antibody to crotoxin.