Methods matter: Comparison of techniques used for sea anemone venom extraction.

The study of cnidarian (coral, sea anemone, and jellyfish) venom provides important evolutionary and ecological insights and unlocks vast opportunities for biodiscovery of novel compounds. The success of the field is dependent on not only the acquisition of sufficient quantities of venom but also the ability to compare venom between species and studies. To date, no direct comparison of the main techniques used to acquire sea anemone venom has been performed to determine the comparability or validity of these methods to yield venom derived from within cnidarian venom apparatus - cnidae. This study aims to compare the venom extracted from a sea anemone via three common methods: isolated cnidae, electrostimulation, and physical manipulation. Using a range of non-targeted proteomic and mass spectrometric techniques, we showed each method yielded distinct differences in both the composition and abundance of components detected for extraction method. Furthermore, few identified components were shared between each of the extraction methods. These results highlight that different venom collection methods yield vastly different results. While further investigation is required, to validate the source of each of the components from within each sample, we argue that sample collection from isolated cnidae is likely to be the most representative of true venom components.