Evaluation of four different immunogens for the production of snake antivenoms

Four different immunogens were used to produce polyvalent antivenom in rabbits to the venoms of Bothrops atrox, Crotalus atrox, Crotalus adamanteus and Crotalus durissus terrificus. The immunogens were: (1) unfractionated mixture of the four crude venoms, and three fractions of the mixture as follows, (2) HPLC gel filtration high (> 50,000) mol. wt fraction, (3) HPLC gel filtration medium (14,000-50,000) mol. wt fraction, and (4) HPLC gel filtration low (< 14,000) mol. wt fraction. The resultant immune sera were compared with commercial antivenom (Wyeth, polyvalent Crotalidae) for total IgG content, ELISA reactivities, patterns of Western blots and ability to neutralize lethal and local hemorrhagic activities of the four venoms. The results indicate that the rabbit antivenoms had significantly higher ELISA reactivity and blotting signals than Wyeth antivenom. However, neither ELISA nor Western blotting signals correlated with the ability of the antivenoms to neutralize the lethal or hemorrhagic activities of the venoms. The protective ability of the antivenoms varied considerably. In general, antivenoms generated by using fractionated venoms as immunogens exhibited greater protective ability than antivenom produced by using the mixture of four venoms as immunogen. Some of the antivenoms provided greater or comparable protective ability for certain venoms when compared to Wyeth antivenom. It appears that the use of certain venom fractions as immunogens is a promising alternative for production of effective antivenoms.