Anti-serine protease monoclonal antibodies from B. jararaca venom: generation of hybridomas

Abstract

Considered by the World Health Organization as a neglected tropical disease of the largest category, snakebite affects about five million people, causes about one hundred thousand deaths annually, and approximately four hundred thousand amputations and other sequelae. In Brazil, about 90% of accidents are caused by snakes of the genus Bothrops; Its venom is a highly complex mixture of proteins, peptides and other factors that, when interacting with human proteins, trigger various effects. The metalloproteases, serine proteases and phospholipase A2, are among the most abundant toxins in botropic venom. The treatment for Bothrops snake envenomation is the administration of the antivenom bothrops, which is produced in horses by administering sublethal doses of a mixture of venoms. Although it is highly effective in the treatment of systemic effects and avoids lethality, the botropic antivenom has some limitations, such as the treatment of local effects, the formation of immune complexes and partial neutralization of toxins belonging to the class of serine proteases. With the evolution of antibody engineering, the addition of a neutralizing monoclonal antibody specific for the serine proteases of the botropic venom to the commercial antivenom may be an improvement strategy for the treatment of envenomation by snakes of the genus Bothrops. In this context, this project aims at the production of hybridomas, the purification and characterization of monoclonal anti-serine antibodies from Bothrops jararaca. In addition, to evaluate in vitro in comparison with commercial botropic antivenom, the neutralizing efficacy of the solution composed of conventional botropic antivenom and monoclonal anti-serine antibodies from Bothrops jararaca.