Reverse-phase high-performance liquid chromatography is commonly employed as a decomplexing strategy in snake venom proteomics. The chromatographic fractions often contain relatively pure toxins that can be assessed functionally for toxicity level through the determination of their median lethal doses (LD50). Further, antivenom efficacy can be evaluated specifically against these venom fractions to understand the limitation of the antivenom as the treatment for snake envenomation. However, methods of toxicity assessment and antivenom evaluation vary across laboratories; hence there is a need to standardize the protocols and parameters, in particular those related to the neutralizing efficacy of antivenom. This chapter outlines the important in vivo techniques and data interpretation that can be applied in the functional study of snake venom proteomes.
1. Examination of the polyacrylamide gel electrophoretic (PAGE) and SDS-PAGE patterns of snake venoms shows that these patterns are useful for species differentiation (and hence identification) for snakes of certain genera but have only limited application for snakes from some other genera, due either to the marked individual variations in the venoms or the lack of marked interspecific differences within the same genus. 2. There is no substantial intersubspecific difference in the electrophoretic patterns of the venoms. 3. In general there are no common characteristics in the electrophoretic patterns of the venom at the generic level because of the wide variations in the electrophoretic patterns of venoms of snakes within the same genus. 4. At the familial level, the venoms of Elapidae exhibited SDS-PAGE patterns distinct from those of Crotalidae.