Venom:antivenom immune complex binding assay using size-exclusion high-performance liquid chromatography (SE-HPLC)
Abstract
The treatment of envenomation with effective snake antivenom immunoglobins has become a critical worldwide health issue. Current methods for testing the effectiveness of new antivenom mixtures in neutralizing venom toxicity/lethality use animal models (e.g. mice). Neutralization of venom toxicity/lethality requires the formation of venom-antivenom immune complexes (though the extent of complex formation in vivo is unknown). Size-exclusion high-performance liquid chromatography (SE-HPLC) is a reproducible quantitative method to characterize venom-antivenom immune complex formation in vitro within a relatively short time. Changes in SE-HPLC elution profiles due to dose-dependent formation of venom-antivenom immune complexes are present for 1) Crotalis atrox (western diamondback rattlesnake) venom and the current antivenom used clinically in North America [FabAV (Ovine); CroFabTM], and 2) Bothrops jararaca venom (Brazil) and Bothropic antivenom [F(ab')2AV (Equine); Brazil]. Changes in profile region areas were fit to a hyperbolic dose-response function to estimate maximum binding and venom/antivenom concentrations at half-maximum binding.
Citation
Sanny, C. G., & Shults, C. A. (2019, Feb. 22). Venom:antivenom immune complex binding assay using size-exclusion high-performance liquid chromatography (SE-HPLC). Poster presented on Research Day at the Oklahoma State University Center for Health Sciences, Tulsa, OK.
Collections
- Research Day 2019 [49]