Fig. 2
Scheme for RP-HPLC fractionation of snake venoms. A considerable number of snake venomic studies have used the chromatographic conditions indicated in the diagram. The venom proteins are separated using an analytical (4.6 × 250 mm, particle diameter of 5 μm) reverse-phase C18 column, eluted at a flow rate of 1 mL/min by a linear gradient of water containing 0.1% of trifluoroacetic acid (TFA) (solution A) and 70% acetonitrile (CNCH3) containing 0.1% TFAa, and the eluate monitored at 215 nm. The timetable for the mixing of these solutions (A, B), and the shape of the gradient (dashed line) are indicated. As an example, the approximate elution regions for some of the common protein components of snake venoms are indicated by colored boxes. This procedure has been applied to venoms of a number of viperid and elapid snakes, helping in the standardization and comparability of results between different laboratories. 3FTx: three-finger toxin; Kunitz: Kunitz-type serine protease inhibitor; PLA2: phospholipase A2; CTL: C-type lectin; SP: serine protease; CRiSP: cystein-rich secretory protein; NGF: nerve-growth factor; VEGF: vascular endothelium growth factor; MP: metalloproteinase; LAAO: L-amino acid oxidase; PDE: phosphodiesterase; 5′-NU: 5′-nucleotidase; HYA: hyaluronidase; PLB: phospholipase B