Improved method for the purification and characterization ingredients of the Naja naja oxiana snake venom

Introduction: The Snake venom toxins are responsible for causing severe pathology and toxicity following envenomation including necrosis, apoptosis, neurotoxicity, myotoxicity, cardiotoxicity, profuse hemorrhage, and disruption of blood homeostasis. However, some snake venom toxins have great potential as drugs for treating human diseases. It is foreseeable that some of peptide or proteins contained in composition of venom can be candidate in use for pharmaceutical purposes, if purified and fully characterized. The purpose of this study is purification proteins composed in Naja Naja Oxiana snake venom and characterization first, secondary and tertiary structures of important ingredients. Method: The Naja Naja Oxiana snake crude venom was obtained from the razi institute as a freeze dried powder. The ingredients of crud venom was purified using gel filtration (sephadex G-50), anion exchange (DEAE sepharose), cation exchange (CM sepharose) and RP HPLC (C18, 150×4.6 mm cloumn). Then desired purified component was characterized. Protein content was determined by Lowry method and toxicity (LD50) via intravenous injection in mice (18-20 gr).Results: In this study five fractions (F1, 2, 3, 4, 5) were isolated by gel filtration of crud venom. The F3 and F4 were toxic. The F3 was selected for further purification. The fraction F3 was purified by anion exchange chromatography on DEAE sepharose. Four anionic fraction collected. First fraction was toxic and named F31. The fraction obtained from anion exchange chromatography was fractionated by cation exchange chromatography and 7 peaks (F311, 312,313,314,315,316,317) achieved in this step. The fractions F314 and F317 were toxic. The fraction F314 showed toxic activity higher than another fractions and finally purified by RP HPLC.Conclusion:

Keywords:

Snake venom , Naja naja oxiana , purification , toxicity