Comparing Traditional and Toxin-Oriented Approaches towards Antivenom Production against <i>Bitis arietans</i> Snake Venom
Felipe Raimondi Guidolin,
Kemily Stephanie de Godoi,
Angela Alice Amadeu Megale,
Cristiane Castilho Fernandes da Silva,
Roberto Tadashi Kodama,
Daniela Cajado-Carvalho,
Leo Kei Iwai,
Patrick Jack Spencer,
Fernanda Calheta Vieira Portaro,
Wilmar Dias da Silva
Affiliations
Felipe Raimondi Guidolin
Immunochemistry Laboratory, Butantan Institute, São Paulo 05503-900, Brazil
Kemily Stephanie de Godoi
Immunochemistry Laboratory, Butantan Institute, São Paulo 05503-900, Brazil
Angela Alice Amadeu Megale
Immunochemistry Laboratory, Butantan Institute, São Paulo 05503-900, Brazil
Cristiane Castilho Fernandes da Silva
Laboratory of Structure and Function of Biomolecules, Butantan Institute, São Paulo 05503-900, Brazil
Roberto Tadashi Kodama
Laboratory of Structure and Function of Biomolecules, Butantan Institute, São Paulo 05503-900, Brazil
Daniela Cajado-Carvalho
Laboratory of Applied Toxinology (LETA), Center of Toxins, Immune-Response and Cell Signaling (CeTICS), Butantan Institute, São Paulo 05503-900, Brazil
Leo Kei Iwai
Laboratory of Applied Toxinology (LETA), Center of Toxins, Immune-Response and Cell Signaling (CeTICS), Butantan Institute, São Paulo 05503-900, Brazil
Patrick Jack Spencer
Nuclear and Energy Research Institute, University of São Paulo, São Paulo 05508-000, Brazil
Fernanda Calheta Vieira Portaro
Laboratory of Structure and Function of Biomolecules, Butantan Institute, São Paulo 05503-900, Brazil
Wilmar Dias da Silva
Immunochemistry Laboratory, Butantan Institute, São Paulo 05503-900, Brazil
Accidents with snakes are responsible for about 32,000 deaths annually in sub-Saharan Africa, caused mostly by snakes from the genus Bitis, in particular Bitis arietans. B. arietans venom is composed of a complex mixture of toxins, mainly metalloproteases, serine proteases, phospholipases, lectins, and disintegrins. In this work, we compared two approaches to anti-B. arietans antivenom production: immunization with crude snake venom (“traditional approach”) and immunization with selected key toxins isolated from the snake venom (“toxin oriented” approach). Fractions from B. arietans venom were isolated by size exclusion chromatography. Crude venom and samples containing serine proteases or metalloproteases were selected for the immunization of BALB/c mice. Anti-B. arietans and anti-serine proteases plasmas showed a similar recognition profile and higher titers and affinity than the anti-metalloproteases plasma. Cross-recognition of other Bitis venoms was observed, but with low intensity. Although the plasma of all experimental groups inhibited the enzymatic activity of B. arietans venom in vitro, in vivo protection was not achieved. Our results have shown limitations in both approaches considered. Based on this, we proposed a model of polyclonal, species-specific, monovalent antivenoms that could be used as a base to produce customizable polyvalent sera for use in sub-Saharan Africa.
