Produção e caracterização de um anticorpo monoclonal contra metaloprotease do veneno da serpente Bothrops atrox

Abstract

Ophidism is a neglected disease that represents a serious public health problem worldwide, with high rates of morbidity and mortality. Among the main snakes, Bothrops genus is responsible for almost 90% of snakebites in Brazil, with Bothrops atrox being the main snake responsible for accidents in the Amazon region. These accidents are known to cause serious systemic and local effects, which can result in amputation of the affected limb, and even lead to the patient death. The symptoms observed in snakebite envenomation occur due to the action of several toxins present in the venom, and among them, metalloproteases stand out. Metalloproteases (SVMPs) are a family of proteins present in Bothrops venom and other snakes of Viperidae family. Commonly classified into three classes, based on their most important protein domains, SVMPs are the main toxins responsible for the hemorrhagic effects caused by bothropic envenomation. The current treatment for such accidents is serum therapy, which, despite being effective, has problems related to production cost, animal ethics and the possibility of adverse reactions. Considering the drawbacks related to serum therapy and the hemorrhagic effect caused by snake venom metalloproteases, the present work aimed to develop a monoclonal antibody capable of neutralizing the hemorrhagic effect of B. atrox venom. For this, the venom was used as an immunogen for the monoclonal antibody production. The selected monoclonal antibody (mAb 4H4D11) is able to recognize bands of approximately 50 and 35 kDa in the venom, and has cross-reactivity with other venoms from Bothrops genus, in addition to Lachesis genus. mAb 4H4D11 partially neutralizes the fibrinogenolytic activity of B. atrox venom in vitro and the hemorrhagic activity in vivo. The results of this work can contribute to the development of new generation antivenoms, aiming to improve the treatment of snakebites.