Purification and characterization of toxic fractions from the Iranian Naja naja Oxiana snake venom

Introduction: Snake venoms contain a vast variety number of pharmacologically active substances. These compounds are of great value in the investigation of important physiologic processes. 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 F4 was selected for further purification. The F4 was purified by anion exchange chromatography and only one fraction resulted in this stage. The fraction obtained from anion exchange chromatography was fractionated by cation exchange chromatography and 5 peaks (F41, 42, 43, 44, 45) achieved in this step. The fractions F42, F43 and F44 were toxic. The fraction F43 showed toxic activity higher than another fractions and finally purified by RP HPLC.Conclusion:

کلیدواژه ها:

Snake ، venom ، Naja naja oxiana ، purification ، toxicity