Volume 90, Issue 3
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645



Envenomation by poisonous animals is a neglected condition according to the World Health Organization (WHO). Antivenoms are included in the WHO Essential Medicines List. It has been assumed that immunoglobulin G (IgG) antivenoms could activate the complement system through Fc and induce early adverse reactions (EARs). However, data in the literature indicate that F(ab') fragments can also activate the complement system. Herein, we show that several batches of IgG and F(ab') antivenoms from the Butantan, Vital Brazil, and Clodomiro Picado Institutes activated the complement classical pathway and induced the production of C3a; however, only those antivenoms from Clodomiro Picado generated C5a. Different protein profiles (IgG heavy chain, protein contaminants, and aggregates) were observed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analyses. Our results show that various antivenoms from different producers are able to activate the classical pathway of the complement system and generate anaphylatoxins, and these findings suggest that factors, such as composition, contaminant proteins, and aggregates, may influence the anticomplementary activity of antivenoms . Therefore, there is a need to further improve antivenom production methods to reduce their anticomplementary activity and potential to cause EARs.


Article metrics loading...

The graphs shown below represent data from March 2017
Loading full text...

Full text loading...



  1. World Health Organization, 2007. Rabies and Envenomings: A Neglected Public Health Issue: Report of a Consultative Meeting. Available at: http://www.who.int/bloodproducts/animal_sera/Rabies.pdf. Accessed February 5, 2013. [Google Scholar]
  2. World Health Organization, 2009. The 17 Neglected Tropical Diseases. Available at: http://www.who.int/neglected_diseases/diseases/en. Accessed February 5, 2013. [Google Scholar]
  3. World Health Organization, 2011. WHO Model List of Essential Medicines. Available at: http://whqlibdoc.who.int/hq/2011/a95053_eng.pdf. Accessed February 5, 2013. [Google Scholar]
  4. World Health Organization, 2010. Guidelines for the Production, Control and Regulation of Snake Antivenom Immunoglobulins. Available at: http://www.who.int/bloodproducts/snake_antivenoms/snakeantivenomguideline.pdf. Accessed February 5, 2013. [Google Scholar]
  5. Lavonas EJ, , 2012. Antivenoms for snakebite: design, function and controversies. Curr Pharm Biotechnol 13: 19801986.[Crossref] [Google Scholar]
  6. Eursakun S, Simsiriwong P, Ratanabanangkoon K, , 2012. Studies on the fractionation of equine antivenom IgG by combinations of ammonium sulfate and caprylic acid. Toxicon 60: 10221029.[Crossref] [Google Scholar]
  7. Cardoso JL, Fan HW, França FO, Jorge MT, Leite RP, Nishioka SA, Avila A, Sano-Martins IS, Tomy SC, Santoro ML, Chudzinski AM, Castro SCB, Kamiguti AS, Kelen EMA, Hirata MH, Mirandola RMS, Theakston RDG, Warrell DA, , 1993. Randomized comparative trial of three antivenoms in the treatment of envenoming by lance-headed vipers (Bothrops jararaca) in São Paulo, Brazil. Q J Med 86: 315325. [Google Scholar]
  8. Isbister GK, Shahmy S, Mohamed F, Abeysinghe C, Karunathilake H, Ariaratnam A, , 2012. A randomised controlled trial of two infusion rates to decrease reactions to antivenom. PLoS One 7: e38739.[Crossref] [Google Scholar]
  9. Morais JF, de Freitas MC, Yamaguchi IK, dos Santos MC, da Silva WD, , 1994. Snake antivenoms from hyperimmunized horses: comparison of the antivenom activity and biological properties of their whole IgG and F(ab')2 fragments. Toxicon 32: 725734.[Crossref] [Google Scholar]
  10. León G, Monge M, Rojas E, Lomonte B, Gutiérrez JM, , 2001. Comparison between IgG and F(ab')2 polyvalent antivenoms: neutralization of systemic effects induced by Bothrops asper venom in mice, extravasation to muscle tissue, and potential for induction of adverse reactions. Toxicon 39: 793801.[Crossref] [Google Scholar]
  11. Otero-Patiño R, Cardoso JLC, Higashi HG, Núñez V, Díaz A, Toro MF, Garcia ME, Sierra A, Garcia LF, Moreno AM, Medina MC, Castañeda N, Silva-Diaz JF, Murcia M, Cardenas SY, Dias da Silva WD, , 1998. A randomized, blinded, comparative trial of one pepsin-digested and two whole IgG antivenoms for Bothrops snake bites in Uraba, Colombia. Trop Med Hyg 58: 183189. [Google Scholar]
  12. Otero-Patiño R, Segura A, Herrera M, Angulo Y, León G, Gutiérrez JM, Barona J, Estrada S, Pereañez A, Quintana JC, Vargas LJ, Gómez JP, Díaz A, Suárez AM, Fernández J, Ramírez P, Fabra P, Perea M, Fernández D, Arroyo Y, Betancur D, Pupo L, Córdoba EA, Ramírez CE, Arrieta AB, Rivero A, Mosquera DC, Conrado NL, Ortiz R, , 2012. Comparative study on the efficacy and safety of two polyvalent, caprylic acid fractionated [IgG and F(ab')2] antivenoms, in Bothrops asper bites in Colombia. Toxicon 59: 344355.[Crossref] [Google Scholar]
  13. Otero R, Gutiérrez JM, Rojas G, Núñez V, Díaz A, Miranda E, Uribe AF, Silva JF, Ospina JG, Medina Y, Toro MF, Garcia ME, León G, Garcia M, Lizano S, De La Torre J, Márquez J, Mena Y, González N, Arenas LC, Puzón A, Blanco N, Sierra A, Espinal ME, Arboleda M, Jiménez JC, Ramírez P, Díaz M, Guzmán MC, Barros J, Henao S, Ramírez A, Macea U, Lozano R, , 1999. A randomized blinded clinical trial of two antivenoms, prepared by caprylic acid or ammonium sulphate fractionation of IgG, in Bothrops and Porthidium snake bites in Colombia: correlation between safety and biochemical characteristics of antivenoms. Toxicon 37: 895908.[Crossref] [Google Scholar]
  14. Herrera M, León G, Segura A, Meneses F, Lomonte B, Chippaux JP, Gutiérrez JM, , 2005. Factors associated with adverse reactions induced by caprylic acid-fractionated whole IgG preparations: comparison between horse, sheep and camel IgGs. Toxicon 46: 775781.[Crossref] [Google Scholar]
  15. Theakston RDG, Warrell DA, Griffiths E, , 2003. Report of a WHO workshop on the standardization and control of antivenoms. Toxicon 41: 541557.[Crossref] [Google Scholar]
  16. Mollnes TE, Hogasen K, De Carolis C, Vaquero E, Nielsen EW, Fontana L, Perricone R, , 1998. High-dose intravenous immunoglobulin treatment activates complement in vivo . Scand J Immunol 48: 312317.[Crossref] [Google Scholar]
  17. García M, Monge M, León G, Lizano S, Segura E, Solano G, Rojas G, Gutiérrez JM, , 2002. Effect of preservatives on IgG aggregation, complement-activating effect and hypotensive activity of horse polyvalent antivenom used in snakebite envenomation. Biologicals 30: 143151.[Crossref] [Google Scholar]
  18. Sarma JV, Ward PA, , 2011. The complement system. Cell Tissue Res 343: 227235.[Crossref] [Google Scholar]
  19. dos Santos MC, D'Império Lima MR, Furtado GC, Colletto GM, Kipnis TL, Dias da Silva W, , 1989. Purification of F(ab')2 anti-snake venom by caprylic acid: a fast method for obtaining IgG fragments with high neutralization activity, purity and yeld. Toxicon 27: 297303.[Crossref] [Google Scholar]
  20. Rojas G, Jiménez JM, Gutiérrez JM, , 1994. Caprylic acid fractionation of hyperimmune horse plasma: description of a simple procedure for antivenom production. Toxicon 32: 351363.[Crossref] [Google Scholar]
  21. Pidde-Queiroz G, Furtado MFD, Filgueiras CF, Pessoa LA, Spadafora-Ferreira M, van den Berg CW, Tambourgi DV, , 2010. Human complement activation and anaphylatoxins generation induced by snake venom toxins from Bothrops genus. Mol Immunol 47: 25372544.[Crossref] [Google Scholar]
  22. Laemmli UK, , 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680685.[Crossref] [Google Scholar]
  23. Morrissey JH, , 1981. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem 117: 307310.[Crossref] [Google Scholar]
  24. Towbin H, Staehelin T, Gordon J, , 1979. Electrophoretic transfer of proteins from acrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 43504354.[Crossref] [Google Scholar]
  25. Tanaka GD, Pidde-Queiroz G, Furtado MFD, van den Berg C, Tambourgi DV, , 2012. Micrurus snake venoms activate human complement system and generate anaphylatoxins. BMC Immunol 13: 4.[Crossref] [Google Scholar]
  26. Solano S, Segura A, León G, Gutiérrez JM, Burnouf T, , 2012. Low pH formulation of whole IgG antivenom: impact on quality, safety, neutralizing potency and viral inactivation. Biologicals 40: 129133.[Crossref] [Google Scholar]
  27. Dall'Acqua WF, Cook KE, Damschroder MM, Woods RM, Wu H, , 2006. Modulation of the effector functions of a human IgG1 through engineering of its hinge region. J Immunol 177: 11291138.[Crossref] [Google Scholar]
  28. Fumia S, Goede JS, Fischler M, Luginbühl A, Frick S, Fodor P, Lutz HU, , 2008. Human F(ab')2-containing immune complexes together with anti-hinge natural antibodies stimulate complement amplification in vitro and in vivo . Mol Immunol 45: 29512961.[Crossref] [Google Scholar]
  29. Zhou W, , 2012. The new face of anaphylatoxins in immune regulation. Immunobiology 217: 225234.[Crossref] [Google Scholar]
  30. Otero R, León G, Gutiérrez JM, Rojas G, Toro MF, Barona J, Rodríguez V, Díaz A, Núñez V, Quintana JC, Ayala S, Mosquera D, Conrado LL, Fernández D, Arroyo Y, Paniaqua CA, López M, Ospina CE, Alzate C, Fernández J, Meza JJ, Silva JF, Ramírez P, Fabra PE, Ramírez E, Córdoba E, Arrieta AB, Warrell DA, Theakston RD, , 2006. Efficay and safety of two whole IgG polyvalent antivenoms, refined by caprylic acid fractionation with or without beta-propiolactone, in the treatment of Bothrops asper bites in Colombia. Trans R Soc Trop Med Hyg 100: 11731182.[Crossref] [Google Scholar]
  31. Stone SF, Isbister GK, Shahmy S, Mohamed F, Abeysinghe C, Karunathilake H, Ariaratnam A, Jacoby-Alner TE, Cotterell CL, Brown SG, , 2013. Immune response to snake envenoming and treatment with antivenom; complement activation, cytokine production and mast cell degranulation. PLoS Negl Trop Dis 7: e2326.[Crossref] [Google Scholar]
  32. Abubakar IS, Abubakar SB, Habib AG, Nasidi A, Durfa N, Yusuf PO, Larnyang S, Garnvwa J, Sokomba E, Salako L, Theakston RD, Juszczak E, Alder N, Warrell DA, Nigeria-UK EchiTab Study Group; , 2010. Randomised controlled double-blind non-inferiority trial of two antivenoms for saw-scaled or carpet viper (Echis ocellatus) envenoming in Nigeria. PLoS Negl Trop Dis 4: e767.[Crossref] [Google Scholar]
  33. Nezlin R, , 2010. Interactions between immunoglobulin G molecules. Immunol Lett 132: 15.[Crossref] [Google Scholar]

Data & Media loading...

  • Received : 12 Oct 2013
  • Accepted : 16 Dec 2013
  • Published online : 05 Mar 2014

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error