Experimental efficacy of IgY antibodies produced in eggs against the venom of the peruvian snake Bothrops atrox

Abstract

Objectives . To develop an immunization protocol in order to produce avian Ig y immunoglobulins against Bothrops atrox Peruvian snake venom and to evaluate its neutralizing capacity. Materials and methods . Six Hy Line Brown hens were immunized each two weeks using 500μg/doses of B. atrox venom in a period of two months. Each week, eggs were collected for Ig y isolation from yolk using two consecutive steps with caprilic acid and ammonium sulfate. Detection of Ig y anti- B. atrox were performed by double immunodiffusion, whereas title and cross-reactivity were analyzed using ELISA and Western Blot technics, respectively. f urthermore, letal dose (DL 50 ) and Medium Effective Dose (DE 50 ) were obtained by Probit analysis. Results . As a result of this protocol, chicken Ig y ’s were obtained in a concentration of 8,5 ± 1,35 mg/yolk mL. DE 50 from avian antivenom was 575 μL/venom mg. Cross-reactivity studies showed Bothrops atrox venom share more commom epitopes with Bothrops brazili (47%) than others Bothrops venoms showing Lachesis muta (19%) and Crotalus durissus (12%) venoms a low crossing reactivity, instead. Conclusions . Using this procedure, we could purify chicken Ig y with a neutralizant capacity of B. atrox venom which is comparable to the antivenom of equine origin and demonstrate its capacity as a immunoanalitical tool to evaluate the cross reactivity with others peruvian snakes.