The application of forensic proteomics to identify an unknown snake venom in a deceased toddler

Proteomics is the global analysis of proteins in a sample, and its methodologies are commonly applied in life science research. Despite its wide applicability however, proteomics is rarely used as a tool in criminal investigations. Here we present a case where the technique provided key evidence in a case that involved the death of a two-year old girl. The defendant was known to keep exotic snakes, including several venomous species, which led the coroner to probe whether there could be snake venom in the blood of the deceased. One major challenge of the investigation was the overwhelming presence of several blood proteins, such as apolipoprotein and complement proteins, which hinders the detection of less abundant analytes. In a counter-acting strategy, a combination of immunodepletion and fractionation methods was used; the sample was then submitted to tandem mass spectrometry for peptide identification. Using this strategy, 15,000 peptides could be sequenced. However, the subsequent challenge was to differentiate between human and snake proteins, given the genetic similarities that are shared by the two vertebrate species. After a thorough bioinformatics search and manual inspection, we found that < 1% of the sequenced peptides could be matched unequivocally to snake proteins, including a well-known venom component, phospholipase A2. This evidence, in part, led to a court-issued search warrant of the defendant's home, followed by his arrest and an eventual guilty plea with formal sentencing to 18 months in prison. The work outlined here is an example of how proteomics technology can help to expand the toolkit for molecular forensics. (C) 2021 Elsevier B.V. All rights reserved.