Produção de anticorpos contra metaloproteases e crotoxina usando antígenos sintéticos e caracterização proteômica e toxicológica do veneno de Bothrops brazili

Abstract

Snakes belonging to the genera Bothrops, Crotalus, Lachesis and Micrurus are responsible for most cases of snakebites in Brazil. Snakes of the genus Crotalus spp and Micrurus spp have neurotoxic venom with high lethality if not neutralized in time. Bothrops and Lachesis venom display proteolytic action, causing permanent damage even after treatment with antiofidic sera. The traditional use of venoms in the immunization of animals for the production of antiofidic sera have drawbacks, since the immunized animals may suffer because of toxic components of the venom, which reduces their life expectancy. Beside, the extensive inter and intraspecific variability in the biochemical composition of snake venoms can affect the neutralizing efficacy of antiofidic sera. Thus, it is crucial to use new technologies to search for and produce alternative non-toxic immunogens that can be used in the production of antiofidic serums. The use of synthetic peptides and multiepitopic proteins that mimic epitopes of the main toxins of each venom would be an alternative for the production of neutralizing antibodies with less impact on experimental animals. This work is presented in the format of three articles, each one containing in a separated chapter of results. Each chapter presents its individualized abstract. The content description of each chapter is shown below: Chapter 1, (Article 1 - Identification of a linear B-cell epitope in the catalytic domain of bothropasin, metalloproteinase from Bothrops jararaca snake venom), this article describes the mapping and synthesis of a linear B cell epitope for bothropasin, capable of to induce neutralizing antibodies. Chapter 2, (Article 2 - Engineered protein containing Crotoxin epitopes induces neutralizing antibodies in immunized rabbits), this article descridbes the production of a multiepitopic recombinant crotoxin protein, capable of inducing neutralizing antibodies. Chapter 3, (Article 3 - Proteomic and toxinological characterization of Peruvian pitviper Bothrops brazili ("jergón shushupe"), venom.), In this article, the components of Bothrops brazili (BbV) venom were analyzed by RP-HPLC, SDS-PAGE and MALDI-TOF/TOF, showing that snake venom metalloproteinases (SVMPs) were the most abundant proteins in the venom of B. brazili. Each article presents its discussion separately. At the end of the third article a general discussion of the three articles is presented.