Isolation and purification of toxic fractions from iranian horned viper (pseudocerastes persicus)

Maedeh sadat Samianifard,1 Ali nazari,2,* Fatemeh tahoori,3

1. Biochemistry Dept. , Razi Vaccine and Serum Research Institute
2. Biochemistry Dept. Razi Vaccine and Serum Research Institute
3. Central Lab Dept ,Razi Vaccine and Serum Research Institute

Abstract


Introduction

Snake venoms are a combination of pharmacologically active molecules, including organic and mineral components, small peptides and proteins. pseudocerastes persicus is a genus of venomous viper widespread in iran. this study was designed to find out the iranian horn viper venom fractions and determining the toxic components.

Methods

The crude venom of pseudocerastes persicus from iran was fractionated using rp-hplc and analyzing them by toxicity test on animals. 0.1 ml of crude venom (1mg/ml) was loaded on to chromatographic column with c18 packing materials with the length 15 cm using agilent 1100. chromatographic separation was performed using the following mobile phases: a - 0.1% trifluoroacetic acid (tfa) in water, b - 0.1% tfa in acetonitrile with gradient elution. all fractions were gathered, a and analyzed for their component by sds-page and toxicity by animal injection.

Results

Horned viper venom was fractionated into 27 peaks with different intensity in absorbance. sds-page analysis revealed the proteins are in a range of 14 to 110 kda, while fractions 12 and 16 were the toxic components.

Conclusion

In brief, determining of toxic fractions of medically important snake venoms can give a strong signal to antivenom manufacturer for making efficient products. this study reveals that iranian horned viper venom includes a varying range of peptide and proteins with two toxic fractions. further experiments are in progress to characterize the real identity of the fractions of this venom.

Keywords

Venom - toxic fraction- pseudocerastes persicus