Abstract
BackgroundSnakebite incidence in southwestern China is mainly attributed to one of the several venomous snakes found in the country, the white-lipped green pit viper Trimeresurus albolabris. Since antivenom produced from horses may cause numerous clinical side effects, the present study was conducted aiming to develop an alternative antivenom antibody (immunoglobulin Y - IgY) from leghorn chickens.MethodsIgY in egg yolk from white leghorn chicken previously injected with T. albolabris venom was extracted by water, precipitated by ammonium sulfate and purified by affinity chromatographic system. IgY was identified by SDS-PAGE, ELISA and Western blot, and its neutralizing assay was conducted on mice.ResultsChickens injected multiple times with T. albolabris venom elicited strong antibody responses, and from their egg yolk IgY was isolated and purified, which exhibited a single protein band on SDS-PAGE and two bands (about 65 and 35 kDa, respectively) under reduced conditions. Immunoblot analysis revealed that these IgY are polyclonal antibodies since they bind with most venom components. In the neutralizing assay, all mice survived while the ratios of IgY/venom reached up to 3.79 (50.0 mg/13.2 mg).ConclusionsIgY antibody response was successfully conducted in white leghorn chicken injected with T. albolabris venom. IgY against T. albolabris venom was obtained for the first time, and it exhibited strong neutralizing potency on mice. These results may lay a foundation for the development of IgY antivenom with clinical applications in the future.
Highlights
Snakebite incidence in southwestern China is mainly attributed to one of the several venomous snakes found in the country, the white-lipped green pit viper Trimeresurus albolabris
To overcome the disadvantages of antivenom made from horse blood, it is essential to attempt other pathways to produce the antibodies against snake venom
Isolation and specific activity of IgY After three steps, IgY from egg yolk was obtained and exhibited a single protein band (180 ~ 200 kDa) through SDS-PAGE. It presented two bands of about 65 kDa and 35 kDa, respectively (Fig. 2), which represented one heavy chain and one light chain of IgY demonstrated by Western blot analysis (Fig. 3)
Summary
Snakebite incidence in southwestern China is mainly attributed to one of the several venomous snakes found in the country, the white-lipped green pit viper Trimeresurus albolabris. Since antivenom produced from horses may cause numerous clinical side effects, the present study was conducted aiming to develop an alternative antivenom antibody (immunoglobulin Y - IgY) from leghorn chickens. In southwestern China, the frequent snakebite cases in rural regions are mainly attributable to Trimeresurus albolabris, Naja naja and Deinagkistrodon acutus. In these areas, antivenom raised in horses are recommended for the treatment of most. We employed chickens to produce antivenom antibodies.
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