Neutralization of the hemolytic activity of the Crotalus durissus terrificus venom by F(ab’2) anti - crotalic PLA2

RODRÍGUEZ, JUAN PABLO; DE MARZI MAURICIO; ACOSTA OFELIA; LEIVA LAURA; MALCHIOIDI EMILIO LUIS

Mar del Plata, Buenos Aires

Congreso; XXXVII Reunión Anual de la Sociedad Argentina de Farmacología Experimental; 2005

We examined the ability of F(ab´)2 from IgG antibodies obtained in rabbits against PLA2, to neutralize whole venom from Crotalus durissus terrificus. PLA2 was isolated from the whole venom by gel filtration chromatography (Sephadex G-75). Specific anti-sera was obtained by subcutaneous and intramuscular inoculation of rabbits with PLA2 (700 μg.ml–1) and Freund adjuvant. IgG antibodies were purified from rabbits anti-sera by FPLC and digested with pepsine to produce F(ab´)2 anti PLA2. Pepsine digestion was developed in a antibodies/enzyme ratio of 50:1 for 18 h at 37 ºC. In order to evaluate the ability of the F(ab´)2 anti-PLA2 to neutralize the activity of the venom, Ouchterlony test, kinetic inhibition test and indirect hemolytic activity test were carried out. The neutralizing capacity of this anti-venom was comparable to that of commercial anti-serum raised against the whole venom. These results strongly suggest that F(ab´)2 against PLA may be considered as an anti-venom that would not produce adverse reactions and/or the inclusion of it as a supplement in polyclonal anti-venoms.