An in Vitro Potency Assay Using Nicotinic Acetylcholine Receptor Binding Works Well with Antivenoms Against Bungarus Candidus and Naja Naja

Overview

Journal Sci Rep

Specialty Science

Date 2018 Jun 28

PMID 29946111

Citations 4

Authors

Kavi Ratanabanangkoon

Pavinee Simsiriwong

Kritsada Pruksaphon

Kae Yi Tan

Bunkuea Chantrathonkul

Sukanya Eursakun

Choo Hock Tan

Affiliations

Soon will be listed here.

Abstract

In order to facilitate/expedite the production of effective and affordable snake antivenoms, a novel in vitro potency assay was previously developed. The assay is based on an antiserum's ability to bind to postsynaptic neurotoxin (PSNT) and thereby inhibit the PSNT binding to the nicotinic acetylcholine receptor (nAChR). The assay was shown to work well with antiserum against Thai Naja kaouthia which produces predominantly the lethal PSNTs. In this work, the assay is demonstrated to work well with antiserum/antivenom against Bungarus candidus (BC), which also produces lethal presynaptic neurotoxins, as well as antivenom against Sri Lankan Naja naja (NN), which produces an abundance of cytotoxins. The in vitro and in vivo median effective ratios (ERs) for various batches of antisera against BC showed a correlation (R) of 0.8922 (p < 0.001) while the corresponding value for the anti-NN antivenom was R = 0.7898 (p < 0.01). These results, together with the known toxin profiles of various genera of elapids, suggest that this in vitro assay could be used with antisera against other species of Bungarus and Naja and possibly other neurotoxic snake venoms worldwide. The assay should significantly save numerous lives of mice and accelerate production of life-saving antivenoms.

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References

. Progress in the characterization of venoms and standardization of antivenoms. WHO Offset Publ. 1981; (58):1-44. View

Gutierrez J, Calvete J, Habib A, Harrison R, Williams D, Warrell D . Snakebite envenoming. Nat Rev Dis Primers. 2017; 3:17079. DOI: 10.1038/nrdp.2017.79. View

Sintiprungrat K, Watcharatanyatip K, Senevirathne W, Chaisuriya P, Chokchaichamnankit D, Srisomsap C . A comparative study of venomics of Naja naja from India and Sri Lanka, clinical manifestations and antivenomics of an Indian polyspecific antivenom. J Proteomics. 2015; 132:131-43. DOI: 10.1016/j.jprot.2015.10.007. View

Tan C, Wong K, Tan K, Tan N . Venom proteome of the yellow-lipped sea krait, Laticauda colubrina from Bali: Insights into subvenomic diversity, venom antigenicity and cross-neutralization by antivenom. J Proteomics. 2017; 166:48-58. DOI: 10.1016/j.jprot.2017.07.002. View

Ranawaka U, Lalloo D, de Silva H . Neurotoxicity in snakebite--the limits of our knowledge. PLoS Negl Trop Dis. 2013; 7(10):e2302. PMC: 3794919. DOI: 10.1371/journal.pntd.0002302. View

Rial A, Morais V, Rossi S, Massaldi H . A new ELISA for determination of potency in snake antivenoms. Toxicon. 2006; 48(4):462-6. DOI: 10.1016/j.toxicon.2006.07.004. View

Harrison R, Hargreaves A, Wagstaff S, Faragher B, Lalloo D . Snake envenoming: a disease of poverty. PLoS Negl Trop Dis. 2009; 3(12):e569. PMC: 2791200. DOI: 10.1371/journal.pntd.0000569. View

Tan N, Wong K, Tan C . Venomics of Naja sputatrix, the Javan spitting cobra: A short neurotoxin-driven venom needing improved antivenom neutralization. J Proteomics. 2017; 157:18-32. DOI: 10.1016/j.jprot.2017.01.018. View

Abe T, Alema S, Miledi R . Isolation and characterization of presynaptically acting neurotoxins from the venom of Bungarus snakes. Eur J Biochem. 1977; 80(1):1-12. DOI: 10.1111/j.1432-1033.1977.tb11849.x. View

10.

Rowan E . What does beta-bungarotoxin do at the neuromuscular junction?. Toxicon. 2000; 39(1):107-18. DOI: 10.1016/s0041-0101(00)00159-8. View

11.

Harris J, Scott-Davey T . Secreted phospholipases A2 of snake venoms: effects on the peripheral neuromuscular system with comments on the role of phospholipases A2 in disorders of the CNS and their uses in industry. Toxins (Basel). 2013; 5(12):2533-71. PMC: 3873700. DOI: 10.3390/toxins5122533. View

12.

Fernandez J, Alape-Giron A, Angulo Y, Sanz L, Gutierrez J, Calvete J . Venomic and antivenomic analyses of the Central American coral snake, Micrurus nigrocinctus (Elapidae). J Proteome Res. 2011; 10(4):1816-27. DOI: 10.1021/pr101091a. View

13.

Williams D, Gutierrez J, Calvete J, Wuster W, Ratanabanangkoon K, Paiva O . Ending the drought: new strategies for improving the flow of affordable, effective antivenoms in Asia and Africa. J Proteomics. 2011; 74(9):1735-67. DOI: 10.1016/j.jprot.2011.05.027. View

14.

Tan K, Tan C, Fung S, Tan N . Venomics, lethality and neutralization of Naja kaouthia (monocled cobra) venoms from three different geographical regions of Southeast Asia. J Proteomics. 2015; 120:105-25. DOI: 10.1016/j.jprot.2015.02.012. View

15.

Tan C, Tan K, Lim S, Tan N . Venomics of the beaked sea snake, Hydrophis schistosus: A minimalist toxin arsenal and its cross-neutralization by heterologous antivenoms. J Proteomics. 2015; 126:121-30. DOI: 10.1016/j.jprot.2015.05.035. View

16.

Maduwage K, Silva A, OLeary M, Hodgson W, Isbister G . Efficacy of Indian polyvalent snake antivenoms against Sri Lankan snake venoms: lethality studies or clinically focussed in vitro studies. Sci Rep. 2016; 6:26778. PMC: 4882578. DOI: 10.1038/srep26778. View

17.

Chippaux J . Snake-bites: appraisal of the global situation. Bull World Health Organ. 1998; 76(5):515-24. PMC: 2305789. View

18.

Laustsen A, Lomonte B, Lohse B, Fernandez J, Gutierrez J . Unveiling the nature of black mamba (Dendroaspis polylepis) venom through venomics and antivenom immunoprofiling: Identification of key toxin targets for antivenom development. J Proteomics. 2015; 119:126-42. DOI: 10.1016/j.jprot.2015.02.002. View

19.

Rosso J, Gutierrez J, Rochat H, Bougis P . Characterization of alpha-neurotoxin and phospholipase A2 activities from Micrurus venoms. Determination of the amino acid sequence and receptor-binding ability of the major alpha-neurotoxin from Micrurus nigrocinctus nigrocinctus. Eur J Biochem. 1996; 238(1):231-9. DOI: 10.1111/j.1432-1033.1996.0231q.x. View

20.

Ramos-Cerrillo B, de Roodt A, Chippaux J, Olguin L, Casasola A, Guzman G . Characterization of a new polyvalent antivenom (Antivipmyn Africa) against African vipers and elapids. Toxicon. 2008; 52(8):881-8. DOI: 10.1016/j.toxicon.2008.09.002. View