In this study, we present high-throughput (HT) venomics,
a novel
analytical strategy capable of performing a full proteomic analysis
of a snake venom within 3 days. This methodology comprises a combination
of RP-HPLC-nanofractionation analytics, mass spectrometry analysis,
automated in-solution tryptic digestion, and high-throughput proteomics.
In-house written scripts were developed to process all the obtained
proteomics data by first compiling all Mascot search results for a
single venom into a single Excel sheet. Then, a second script plots
each of the identified toxins in so-called Protein Score Chromatograms
(PSCs). For this, for each toxin, identified protein scores are plotted
on they-axis versus retention times of adjacent
series of wells in which a toxin was fractionated on thex-axis. These PSCs allow correlation with parallel acquired intact
toxin MS data. This same script integrates the PSC peaks from these
chromatograms for semiquantitation purposes. This new HT venomics
strategy was performed on venoms from diverse medically important
biting species;Calloselasma rhodostoma,Echis ocellatus,Naja pallida,Bothrops asper,Bungarus multicinctus,Crotalus atrox,Daboia russelii,Naja naja,Naja nigricollis,Naja mossambica, andOphiophagus hannah. Our data suggest that high-throughput
venomics represents a valuable new analytical tool for increasing
the throughput by which we can define venom variation and should greatly
aid in the future development of new snakebite treatments by defining
toxin composition.