Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Review
. 2017 May 13;9(5):163.
doi: 10.3390/toxins9050163.

Preclinical Evaluation of the Efficacy of Antivenoms for Snakebite Envenoming: State-of-the-Art and Challenges Ahead

José María Gutiérrez  1 , Gabriela Solano  2 , Davinia Pla  3 , María Herrera  4   5 , Álvaro Segura  6 , Mariángela Vargas  7 , Mauren Villalta  8 , Andrés Sánchez  9 , Libia Sanz  10 , Bruno Lomonte  11 , Guillermo León  12 , Juan J Calvete  13
Affiliations
Review

Preclinical Evaluation of the Efficacy of Antivenoms for Snakebite Envenoming: State-of-the-Art and Challenges Ahead

José María Gutiérrez et al. Toxins (Basel). .

Abstract

Animal-derived antivenoms constitute the mainstay in the therapy of snakebite envenoming. The efficacy of antivenoms to neutralize toxicity of medically-relevant snake venoms has to be demonstrated through meticulous preclinical testing before their introduction into the clinical setting. The gold standard in the preclinical assessment and quality control of antivenoms is the neutralization of venom-induced lethality. In addition, depending on the pathophysiological profile of snake venoms, the neutralization of other toxic activities has to be evaluated, such as hemorrhagic, myotoxic, edema-forming, dermonecrotic, in vitro coagulant, and defibrinogenating effects. There is a need to develop laboratory assays to evaluate neutralization of other relevant venom activities. The concept of the 3Rs (Replacement, Reduction, and Refinement) in Toxinology is of utmost importance, and some advances have been performed in their implementation. A significant leap forward in the study of the immunological reactivity of antivenoms against venoms has been the development of "antivenomics", which brings the analytical power of mass spectrometry to the evaluation of antivenoms. International partnerships are required to assess the preclinical efficacy of antivenoms against snake venoms in different regions of the world in order to have a detailed knowledge on the neutralizing profile of these immunotherapeutics.

Keywords: antivenomics; antivenoms; neutralization tests; preclinical efficacy; snake venoms; the 3Rs.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest. The founding sponsors had no role in the design of the study, in the writing of the manuscript, and in the decision to publish the review.

Figures

Figure 1
Figure 1
Most relevant toxic activities induced by venoms of different snake groups that need to be considered in the preclinical evaluation of antivenoms. Venoms of many elapid species, and few viperids, mainly induce neurotoxicity, which is assessed by lethality. Sea snake venoms induce both neurotoxicity and systemic myotoxicity. The main effect in envenomings by spitting cobras is cutaneous necrosis, which has to be evaluated in addition to lethality. Several Australian terrestrial elapids, as well as few rattlesnake venoms, induce neurotoxicity, myotoxicity and consumption coagulopathy, which depends on their coagulant enzymes. Envenomings by many viperid species cause, in addition to lethality, local tissue damage (hemorrhage and myotoxicity), and systemic effects associated with bleeding and coagulopathy. Other pathophysiological alterations that would need to be considered in the preclinical evaluation of antivenoms are thrombocytopenia, platelet hypoaggregation, acute kidney injury, and systemic vascular capillary leakage syndrome. Reproduced from [46], copyright 2013 Elsevier.
Figure 2
Figure 2
Possibilities for implementing the 3Rs in antivenom preclinical assessment. Some examples of interventions that have been developed or are being implemented in Toxinology laboratories to replace, reduce and refine the assays using animals are described.
Figure 3
Figure 3
Cartoon of the “second generation” antivenomics workflow [127]. Panels (A) and (B) illustrate, respectively, the generation of the immunoaffinity (a) and control antibody (d) columns, and the steps of the antivenomic protocol to assess which toxins show immunoreactivity towards the immobilized antivenom molecules (c) and which do not bind to the immunoaffinity column (b). Mock matrix and control IgG columns, run in parallel to the immunocapture experiment, serve as specificity controls. Panel (C) displays three immunoaffinity experiments using venoms of snakes from different families: antivenomic analysis of D. typus (Colubridae: Colubrinae) venom against CroFab™ antivenom (left) [134], antivenomic analysis of P. papuanus (Elapidae) venom against Australian antivenom (middle) [132], and antivenomic analysis of B. diporus (Viperidae: Crotalinae) venom against Butantan pentabothropic antivenom (right) [135]. Chromatograms labeled “a” display reference RP-HPLC separation of the venom proteins. Major protein classes identified in the different chromatographic fractions are highlighted (3FTx, three-finger toxin; PLA2, phospholipase A2; CRISP, cysteine-rich secretory protein; SVSP, snake venom serine proteinase; PIII-SVMP, snake venom metalloproteinase of class PIII; CTL, C-type lectin-like molecule; LAO, L-amino acid oxidase; VAP, vasoactive peptide; SVMPi, tripeptide inhibitor of SVMPs). Chromatograms “b” and “c” display, respectively, reverse-phase separations of the immunocaptured and the non-bound column fractions recovered from the immunoaffinity columns. Chromatograms “d” and “e” show, respectively, reverse-phase HPLC separations of the venom components recovered in the bound fractions of mock matrix and control IgG columns.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Chippaux J.P. Snake-bites: Appraisal of the global situation. Bull. World Health Organ. 1998;76:515–524. - PMC - PubMed
    1. Gutiérrez J.M., Theakston R.D.G., Warrell D.A. Confronting the neglected problem of snake bite envenoming: The need for a global partnership. PLoS Med. 2006;3:e150. doi: 10.1371/journal.pmed.0030150. - DOI - PMC - PubMed
    1. Kasturiratne A., Wickremasinghe A.R., de Silva N., Gunawardena N.K., Pathmeswaran A., Premaratna R., Savioli L., Lalloo D.G., de Silva H.J. The global burden of snakebite: A literature analysis and modelling based on regional estimates of envenoming and deaths. PLoS Med. 2008;5:e218. doi: 10.1371/journal.pmed.0050218. - DOI - PMC - PubMed
    1. Calmette A. Contribution a l’etude du venin des serpents. Immunisation des animaux et traitment de l’envenimation. Ann. Inst. Pasteur. 1894;8:275–291.
    1. Phisalix C., Bertrand G. Sur la propriete antitoxique du sang des animaux vaccines contre le venin de vipere. C. R. Soc. Biol. 1894;46:111–113.

Publication types

LinkOut - more resources