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. 2014 Aug 19;6(8):2471-82.
doi: 10.3390/toxins6082471.

Immunological cross-reactivity and neutralisation of European viper venoms with the monospecific Vipera berus antivenom ViperaTAb

Affiliations

Immunological cross-reactivity and neutralisation of European viper venoms with the monospecific Vipera berus antivenom ViperaTAb

Nicholas R Casewell et al. Toxins (Basel). .

Abstract

Medically important cases of snakebite in Europe are predominately caused by European vipers of the genus Vipera. The mainstay of snakebite therapy is polyclonal antibody therapy, referred to as antivenom. Here we investigate the capability of the monospecific V. berus antivenom, ViperaTAb®, to cross-react with, and neutralise lethality induced by, a variety of European vipers. Using ELISA and immunoblotting, we find that ViperaTAb® antibodies recognise and bind to the majority of toxic components found in the venoms of the Vipera species tested at comparably high levels to those observed with V. berus. Using in vivo pre-clinical efficacy studies, we demonstrate that ViperaTAb® effectively neutralises lethality induced by V. berus, V. aspis, V. ammodytes and V. latastei venoms and at much higher levels than those outlined by regulatory pharmacopoeial guidelines. Notably, venom neutralisation was found to be superior to (V. berus, V. aspis and V. latastei), or as equally effective as (V. ammodytes), the monospecific V. ammodytes "Zagreb antivenom", which has long been successfully used for treating European snake envenomings. This study suggests that ViperaTAb® may be a valuable therapeutic product for treating snakebite by a variety of European vipers found throughout the continent.

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Figures

Figure 1
Figure 1
In vitro ELISA cross-reactivity of ViperaTAb® and Zagreb antivenoms to a variety of European viper venoms. The 50% binding value represents the amount of antivenom (in ng) required to bind half of the test venom on the microtitre plate in the ELISA experiment. Thus, the lower the 50% value, the greater the binding capacity of the antivenom. Bars for the venoms used to raise the antibodies present in each antivenom are highlighted by bold borders (V. berus for ViperaTAb®; V. ammodytes for Zagreb). Error bars represent standard deviation.
Figure 2
Figure 2
The protein profiles of four Vipera venoms and their immunological cross-reactivity with ViperaTAb® and Zagreb antivenoms. (A) One dimensional SDS-PAGE gel of four Vipera venoms stained with Coomassie Blue R-250 and their immunological cross-reactivity with (B) the monospecific V. berus antivenom ViperaTAb® and (C) the monospecific V. ammodytes Zagreb antivenom.
Figure 3
Figure 3
Relative in vivo potencies of ViperaTAb® and Zagreb antivenoms against venoms from four species of the genus Vipera. (A) Lethal dose 50 (LD50) values for each of the four Vipera venoms studied; (B) Effective dose 50 (ED50) values detailing the relative potency of ViperaTAb® and Zagreb antivenoms. The effective dose is the amount of antivenom antibodies required to protect 50% of a population of mice from the lethal effects of five LD50 doses of each venom. Error bars represent 95% confidence intervals.

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