Immunochemical Studies on Fluoresceinthiocarbamyl and Reduced S-Carboxymethylated Cobrotoxin*

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Abstract
The exhaustive fluorescein thiocarbamylation of cobrotoxin through its free amino groups resulted in decrease in lethality to one-sixth that of native toxin without affecting the antigenic specificity. This suggests that the antigenic sites of cobrotoxin are different from the active site(s) of toxicity.Reduced S-carboxymethylated cobrotoxin was completely devoid of lethality and antigenic activity. This indicates that the intact disulfide bonds which maintain the specific secondary structure are essential for the venom toxicity as well as antigenicity.No significant differences were observed between the immunochemical properties of cobrotoxin and fluoresceinthiocarbamyl cobrotoxin as indicated by the results obtained from the precipitin reactions, immunodiffusion and immunoelectrophoresis, reactivities with Fab fragments of the purified antibody, and measurement of the antibody-binding sites in each antigen molecule. Fluoresceinthiocarbamyl cobrotoxin is a good antigen for the production of anti-cobrotoxin antibody.

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