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Golchinfar,, F., Madani,, R., Emami,, T., Zoulfagharian,, H., Zare,, A., Mouhammadpour,, N. (2016). Designing a competitive ELISA for evaluation of anti-snake venom serum potency. , 29(1), 9-16. doi: 10.22034/vj.2016.105724
F. Golchinfar,; R. Madani,; T. Emami,; H. Zoulfagharian,; A. Zare,; N. Mouhammadpour,. "Designing a competitive ELISA for evaluation of anti-snake venom serum potency". , 29, 1, 2016, 9-16. doi: 10.22034/vj.2016.105724
Golchinfar,, F., Madani,, R., Emami,, T., Zoulfagharian,, H., Zare,, A., Mouhammadpour,, N. (2016). 'Designing a competitive ELISA for evaluation of anti-snake venom serum potency', , 29(1), pp. 9-16. doi: 10.22034/vj.2016.105724
Golchinfar,, F., Madani,, R., Emami,, T., Zoulfagharian,, H., Zare,, A., Mouhammadpour,, N. Designing a competitive ELISA for evaluation of anti-snake venom serum potency. , 2016; 29(1): 9-16. doi: 10.22034/vj.2016.105724

Designing a competitive ELISA for evaluation of anti-snake venom serum potency

تحقیقات دامپزشکی و فرآورده‌های بیولوژیک
Article 2, Volume 29, Issue 1, March 2016, Pages 9-16    PDF (749.7 K)
Document Type: Full Research Paper
DOI: 10.22034/vj.2016.105724
Authors
F. Golchinfar,* 1; R. Madani,2, 3; T. Emami,1; H. Zoulfagharian,4; A. Zare,5; N. Mouhammadpour,6
1Dept. of Biochem & Proteomics, Razi Vaccine and Serum Research Institute, Karaj, IR Iran.
2Proteomics and Biochemistry Department, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.
3Department of Microbiology, School of Specialized Veterinary Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.
4Dept. of Serum & Proteomics, Razi Vaccine and Serum Research Institute, Karaj, IR Iran.
5Dept. of Venomous Animals and Antivenom Production, Razi Vaccine and Serum Research Institute, Karaj, IR Iran.
6Dept. of Venomous Animals and Antivenom Production, Razi Vaccine and Serum Research Institute, Karaj, IR Iran.
Abstract
The venom of Naja naja oxiana, Vipera lebetina, Vipera albicornouta, Echis carinatus, Pseadocerastus persicus and Agkistrodon halys snakes are used to determine the potency of the polyvalent sera in neutralizing the venom. LD50 value per mouse was determined 6.5 μg. Neutralizations methods for obtaining potency of anti venom are competitive ELISA (Enzyme linked immunosorbent assay) and in vivo assay (ED50). Both of these tests were performed to estimate the serum potency in 15 samples of hyper-immune equines. Results of competitive ELISA showed that 1/12000 diluted serum can inhibits 50 percent of antigens. Competitive ELISA was compared with current biological assay ED50. Significant correlations between ELISA titers and values of ED500 at level of P≤ 0.01 and r=0.95 was observed, that indicates competitive ELISA can estimate antibody neutralizing capacity of the serum as well as the in vivo assay. The results of present study shows C-ELISA that measures Ag-Ab complex can used as a suitable replacement method for lethal neutralizing in vivo method.
Keywords
ELISA; Snake venom; Potency; Neutralization
References

1.Alape-Girón, A Miranda-Arrieta, K, Cortes-Bratti,X Stiles,B and Gutiérrez.j. 1997. "A comparison of in vitro methods for assessing the potency of therapeutic antisera against the venom of the coral snake Micrurus nigrocinctus.," Toxicon, vol. 35, pp. 573- 581.
2. Art Carter, Wyeth-Ayerst Research, Chazy, NY.1991. Using the Spearman-Karber method to stimate the ED50, The Best contributed paper. pp.1120-1125.
3. Carpent, A. B. 1992. "Enzyme link immunosorbent assay," ClinicalLaboratory Immunology ,vol.1, pp. 20–29.
4. Crowther, J. R. 2001. the Elisa guide book: human press , NewJersey.
5. Crowther, J. R. 2002. "The ELISA Guide book, method in molecular biology.
6. Dolimbek, B. Z., Atassi, M. Z and. Salikhov, S. I. 1998."Presynaptic and postsynaptic neurotoxins. Investigation of the structures of the immune recognition sections." Chemistry of Natural Compounds, vol. 34, pp. 15- 28.
7. Dowling, T. P. 1978. "Primary care--the Cinderella of health care and of nursing too," J N Y State Nurses Assoc, vol. 9, pp. 21-4,
8. Engrall, E. and Perlman, P. 1971. "Enzyme–linked immunosorbent assay (ELISA), Quantitative assay of Immunoglobulin G,". Immunochemistry, vol. 8, pp. 871-874.
9. Hatanaka, H. Oka, M Kohda, D. Tate, S. Suda, A. Tamiya, N. and Inagaki, F. 1994. "Tertiary structure of erabutoxin b in aqueous solution as lucidated by two-dimensional nuclear magnetic resonance.," J Mol Biol., vol. 240, pp. 155-166.
10. Hock.B, Rahman.M, Rauchalles S, Dankwardt A, Seifert, M, Haindl S , and Kramer K, 1999. "Stabilisation of immunoassays and receptor assays," Journal of Molecular Catalysis B: Enzymatic vol. 7, pp. 115–124.
11. Karlsson, E. 1979 "Chemistry of protein toxins in snake venoms. Snake venoms," in Handbook of experimental pharmacology. vol. 52, C.-Y. e. Lee, Ed., ed Berlin: Springer Verlag, , pp. 159-212.
12. Kumar, B. K. Nanda, S. S. Venkateshwarlu, P., kumar, Y. K. and Jadhav, R. T. 2010 . "ANTISNAKE VENOM SERUM (ASVS)," Indian journal of pharmaceutical and biological research. vol. 1, pp. 76-89.
13. Lequn, R. 2005."Enzyme immunoassay (EIA)/ Enzyme linked immunosorbent assay (ELISA)," Clin, Chem, vol. 51, pp. 2415-2418.
14. Maria, W. S., Pacheco, B G., Barbosa, C F Velarde, D T, and Chávez- Olórtegui, C. 2001. Determination of the neutralizing potency of horse antibothropic and anticrotalic antivenoms in blood samples collected on filter paper. Toxicon vol 39, No,10, pp1607-9.
15. Mordvintsev, D. Y. Polyak, Y. L. Levtsova, O. V. Tourleigh, Y. V. Kasheverov, I. E. Shaitan, K. V. Utkin, Y. N and. Tsetlin, V. I .2005 "A model for short alpha-neurotoxin bound to nicotinic acetylcholine receptor from Torpedo californica: comparison with long-chain alpha- neurotoxins and alpha-conotoxins," Comput Biol Chem, vol. 29, pp. 398- 411.
16. Mukherjee, A. K. and. Maity, C. R. 2002."Biochemical composition, lethality and pathophysiology of venom from two cobras-- Naja naja and N. kaouthia," Comp Biochem Physiol B Biochem Mol Biol, vol. 131, pp. 125-32.
17. Nasiripourdori, A. Ranjbar, B.. Naderi-Manesh, H Mehrnejad, F. Soofian, S. Sadeghi, G. and Kolahian, S. 2008."Structural-functional studies of peptides derived from a long-chain snake neurotoxin Naja naja oxiana," Physiology and Pharmacology, vol. 12, pp. 209-220.
28. Nielsen, K. Smith, P. yu, W. Celmgren, L. Nicoletti, P. Perez, B. Bermudez, R. and Renteria, T. 2007." Second generation Competitive enzyme immunoassay for detection of bovine antibodies to brucella abortus in bulk milk samples. Veterinary of Microbiology. vol. 82, pp. 55-60.
19. Nirthanan, S. and Gwee, M. C. E. 2004."Three-Finger -Neurotoxins and the Nicotinic Acetylcholine Receptor,Forty Years On" Journal of Pharmacological Sciences, vol. 94, pp. 1-17.
20. Rial, A. Morais V., Rossi, S. and Massaldi, H. 2006 "A new ELISA for determination of potency in snake antivenoms," Toxicon, vol. 48, pp. 462-6.
21. Raweeritha, R. and Ratanabanangkoon, K. 2005. "Immunochemical and biochemical comparisons of equine monovalent and polyvalent snake antivenoms," Toxicon, vol. 45, pp. 369–375,.
22. Rowel, V. 1992. "Nunc Guid tosalid phase," Nunc Brand product, pp. 5-35.
23. Rungsiwongse, J. and Ratanabanangkoon, K. 1991."Development of an ELISA to assess the potency of horse therapeutic antivenom against Thai cobra venom," J Immunol Methods, vol. 136, pp. 37-43.
24. Sells, P. G. 2003."Animal experimentation in snake venom
research and in vitro alternatives," Toxicon, vol. 42, pp. 115-33.
25.Shashidharamurthy, R. Kemparaju , K. A 2006. Neurotoxic phospholipase A2 variant: Isolationand characterization from eastern regionalIndian cobra (Naja naja) venom, Toxicon 47, pp.727–733.
26. Solano, G. Segura, A. Herrera, M. Gómez, A. Villalta, M. Gutiérrez, J. and León, G. 2010. "Study of the design and analytical properties of the lethality neutralization assay used to estimate antivenom potency against Bothrops asper snake venom.," Biologicals., vol. 38, pp. 577-585.
27. Stiles, B. G. Sexton, F. W. Guest, S. B. Olson, M. A. and Hack, D. C.1994."Characterization of monoclonal antibodies against Naja naja oxiana neurotoxin I," Biochem J, vol. 303 ( Pt 1), pp. 163-70.
28. Tsetlin, V. I. and Hucho, F. 2004 "Snake and snail toxins acting on nicotinic acetylcholine receptors: fundamental aspects and medical applications," FEBS Lett, vol. 557, pp. 9-13.

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