Development of high-throughput screening assays for profiling snake venom phospholipase A2 activity after chromatographic fractionation

Abstract

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Varespladib Inhibits the Phospholipase A2 and Coagulopathic Activities of Venom Components from Hemotoxic Snakes

“…The PLA 2 activity assay was carried out according to the method recently reported by Still et al [30] using cresol red as a pH indicator. The PLA 2 assay monitors the decrease in pH caused by the enzymatic conversion of L-a-Phosphatidylcholine to fatty acids.…”

A Spider Toxin Exemplifies the Promises and Pitfalls of Cell-Free Protein Production for Venom Biodiscovery

“…Even so, it is important to point out that upscaling by combining multiple batches may also increase costs, so the economic benefits of cell-free production would need to be reassessed. The small amounts of protein synthesized in cell-free systems could also be isolated and enriched by chromatography, as shown by the micro-fractionation of venom components [ 33 , 34 , 35 , 36 , 37 ]. However, this would also add more experimental steps, thus eliminating one of the major advantages of cell-free expression: the rapid and simple production method.…”

Small Molecule Drug Discovery for Neglected Tropical Snakebite

“…This is particularly pertinent when considering that current drug candidates predominately target just two of the major toxin families found in venom, PLA 2 s and SVMPs [13], justifying the need for expanding the chemical space explored against these toxins and other snakebite drug targets to strengthen the possibility of developing a safe and globally effective (pan-species) snakebite drug therapy. There are many validated in vitro models available for key venom toxin families that, due to their basic cellular or biochemical format, will make them amenable to high-throughput screening (HTS) to rapidly expand the chemical space available for drug discovery [29,[51][52][53]. Once such in vitro models are adapted and validated for HTS, appropriate drug libraries should be selected for testing.…”