Merck

Bothropic antivenom based on monoclonal antibodies, is it possible?

Toxicon : official journal of the International Society on Toxinology (2013-06-05)
Thiago S Frauches, Jorge H Petretski, Andrea C V Arnholdt, Elena B Lasunskaia, Eulógio C Q de Carvalho, Thereza L Kipnis, Wilmar D da Silva, Milton M Kanashiro
ABSTRACT

Neutralizing monoclonal antibodies against three major toxic components of Bothrops atrox venom were produced and tested. The mAbs against phospholipase A2, hemorrhagic metalloprotease, and thrombin-like enzymes were produced in large amounts and purified with caprylic acid followed by ammonium sulfate precipitation. Purified mAbs were analyzed by SDS-PAGE and their ability to neutralize the respective toxins was tested. Five Swiss mice were injected i.p. with 13.5 mg of pooled mAbs and challenged via s.c. route with venom. Survival rate was recorded for the next 48 h. All mice treated and challenged with venom survived, whereas only one mouse in the control group survived. Bleeding time in mice treated with mAbs was similar to that observed in control mice. Our results show that monoclonal antibodies neutralized the lethal toxicity of Bothrops venom and indicate that there is a reasonable possibility of developing antivenoms based on humanized mAbs to treat victims of venomous animals in the future.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ammonium-14N2 sulfate solution, 40 wt. % in H2O, 99.99 atom % 14N
Sigma-Aldrich
Octanoic acid, ≥99%
Sigma-Aldrich
Ammonium sulfate-14N2 solution, 40 wt. % in H2O, 99.99 atom % 14N
Sigma-Aldrich
Octanoic acid, natural, ≥98%, FG
Sigma-Aldrich
Octanoic acid, ≥98%, FG
Sigma-Aldrich
Ammonium sulfate, suitable for plant cell culture, ≥99.0%
Sigma-Aldrich
Ammonium sulfate, BioXtra, ≥99.0%
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Ammonium sulfate, for molecular biology, ≥99.0%
Sigma-Aldrich
Ammonium sulfate, BioUltra, ≥99.0% (T)
Supelco
Ammonium sulfate, analytical standard, for Nitrogen Determination According to Kjeldahl Method, traceable to NIST SRM 194
Sigma-Aldrich
Ammonium sulfate, anhydrous, Redi-Dri, ReagentPlus®, ≥99.0%
Sigma-Aldrich
Ammonium sulfate, anhydrous, free-flowing, Redi-Dri, ACS reagent, ≥99.0%
Sigma-Aldrich
Ammonium sulfate, ReagentPlus®, ≥99.0%
Sigma-Aldrich
Ammonium sulfate, ACS reagent, ≥99.0%
Caprylic acid, European Pharmacopoeia (EP) Reference Standard
Supelco
Caprylic Acid (Octanoic Acid), Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Ammonium sulfate, 99.999% trace metals basis