Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

Optimization of purification procedure for horse F(abÂ´)2 antivenom against Androctonus crassicauda (Scorpion) venom

Azadeh Taherian¹ , Mohammad Fazilati², Ahmad Taghavi Moghadam³, Hamid Tebyanian⁴

¹Department of Biology, Payame Noor University; ²Department of Biochemistry, Payame Noor University, P.O.Box 19395-4697; ³Razi Serum and Vaccine Research Institute, Ahwaz Branch, Agricultural Research, Education and Extension Organization (AREEO); ⁴Research Center for Prevention of Oral and Dental Disease, Baqiyatallah University of Medical Sciences, Tehran, Iran.

For correspondence:- Azadeh Taherian Email: azadeh.taherian@yahoo.com Tel:+98 9166098242

Accepted: 25 February 2018 Published: 31 March 2018

Citation: Taherian A, Fazilati M, Moghadam AT, Tebyanian H. Optimization of purification procedure for horse F(abÂ´)2 antivenom against Androctonus crassicauda (Scorpion) venom. Trop J Pharm Res 2018; 17(3):409-414 doi: 10.4314/tjpr.v17i3.4

© 2018 The authors.
This is an Open Access article that uses a funding model which does not charge readers or their institutions for access and distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) and the Budapest Open Access Initiative (http://www.budapestopenaccessinitiative.org/read), which permit unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited..

Abstract

Purpose: To immunize antiserum of horse with Androctonus crassicauda scorpion venom in order to achieve an antivenom with higher purity by combined caprylic acid/ammonium sulfate.

Methods: The optimum pH to terminate enzymatic digestion was evaluated. Purification was performed by various combinations of caprylic acid (0 to 2.5 mL %) and ammonium sulfate (0 to 20 g %) at 25, 30 and 37 ºC. The effects of three factors (caprylic acid, ammonium sulfate and temperature) were evaluated based on precipitation of non-immunoglobulin proteins. Antivenom purity was evaluated by determining the concentration of desired soluble protein and undesired albumin, as well as by turbidity and titration.

Results: The results showed that the optimum pH for inhibition of enzyme activity and precipitation of impurities was 4.8. SDS-PAGE revealed that the highest impurity precipitation and lowest protein aggregation was occurred at the combination of 1.5 mL % caprylic acid and 10 g % ammonium sulfate at 37 ºC.

Conclusion: The modified method of purification significantly decreases turbidity, albumin impurity concentration and processing time but increased antibody titer and purity of antivenom. Therefore, it is a potentially suitable method for purifying antivenom in commercial production

Keywords: Antivenom, Scorpion, Androctonus crassicauda, Purification, Albumin, Antiserum