A one-step isolation procedure for phospholipase A2-free cobra venom factor by fast protein liquid chromatography.
A very rapid and efficient procedure for isolation of cobra venom factor (CoF) from Naja naja venom is presented. The method is based on Mono Q anion exchange chromatography on a system for fast protein liquid chromatography (FPLC). CoF was eluted by a buffer of pH 7.4 at 280 mM salt. A purification of 33.7 X was reached with a yield of at least 27%. Contamination with phospholipase was under the detection limit of a sensitive radiometric assay (less than 25 ppm), while the starting material contained 5%. The preparation displays high C-depleting activity in vivo.
D000818:Animals; D002852:Chromatography, Ion Exchange / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D003169:Complement Inactivator Proteins / Q000302:isolation & purification; D004546:Elapid Venoms / Q000302:isolation & purification*; D008970:Molecular Weight; D010740:Phospholipases / Q000302:isolation & purification*; D010741:Phospholipases A / Q000302:isolation & purification*; D054467:Phospholipases A2
D003169:Complement Inactivator Proteins; D004546:Elapid Venoms; C016310:cobra venom factor; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2