Production of policlonal antibodies against three Peruvian snake venoms and its evaluation by immunoenzymatic methods

Abstract

In Peru, bites caused by snakes of genus Bothrops and Lachesis are the most important recorded, and the identification of responsible snake is a poorly attended issue, so it is highly recommended to develop immunochemical methods for this purpose. The aim of this research was to produce polyclonal antibodies against venoms of B. atrox, B. barnetti and L. muta, and to evaluate its cross-reactivity by immunoenzymatic methods. To reach this objective, an immunization protocol using New Zealand white rabbits (2 kg) was followed, using complete venoms of three snake species. Venom content was analyzed by SDS-PAGE, while IgGs obtained at the final of protocol were analyzed by ELISA and Western Blot. As part of the results, the immunization schedule allowed producing antibodies from day 20 and sera title was 64000. Cross-reactivity values were 25% and 4% using anti-B. atrox serum against B. barnetti and L. muta venoms, respectively. In Western Blot experiments, reactivity bands were obtained in a range of 14.3 and 97 kDa. In conclusion, the venoms in study are highly immunogenic and produced IgGs could be used to evaluate cross-reactivity and to elaborate a kit for rapid diagnosis of envenomation by these snake’s bites.