Qualitative Profiling of Venom Toxins in the Venoms of Several Species Using High-Throughput Venomics and Coagulation Bioassaying.

Envenoming resulting from snakebites is recognized as a priority neglected tropical disease by The World Health Organization. The genus, consisting of different pitviper species, is considered the most medically significant taxa in Central and South America. Further research into venom composition is important to aid in the development of safer and more effective snakebite treatments. In addition, the discovery of toxins that could potentially be used for medical or diagnostic purposes is of interest to the pharmaceutical industry. This study aimed to employ high-throughput (HT) venomics to qualitatively analyze venom composition while utilizing coagulation bioassays for identifying coagulopathic toxins and characterizing coagulopathic activity in various venoms. Using the recently demonstrated HT venomics workflow in combination with post-column coagulopathic bioassaying, focus was placed at anticoagulant toxins. Well-known procoagulant toxins were also investigated, taking into account that using the HT venomics workflow, procoagulant toxins are especially prone to denaturation during the reversed-phase chromatographic separations performed in the workflow. The findings revealed that the venoms of and harbored procoagulant toxins, whereas those of and contained both procoagulant and anticoagulant toxins. In general, anticoagulation was associated with phospholipases As, while procoagulation was associated with snake venom metalloproteinases and snake venom serine proteases. These results showed the identification of coagulopathic venom toxins in the venoms analyzed using multiple analytical methods that complement each other. Additionally, each venom underwent qualitative characterization of its composition.