PT - JOURNAL ARTICLE AU - Jenson, Sarah C. AU - Chu, Fanny AU - Barente, Anthony S. AU - Crockett, Dustin L. AU - Lamar, Natalie C. AU - Merkley, Eric D. AU - Jarman, Kristin H. TI - MARLOWE: Taxonomic Characterization of Unknown Samples for Forensics Using <em>De Novo</em> Peptide Identification AID - 10.1101/2024.09.30.615220 DP - 2024 Jan 01 TA - bioRxiv PG - 2024.09.30.615220 4099 - http://biorxiv.org/content/early/2024/09/30/2024.09.30.615220.short 4100 - http://biorxiv.org/content/early/2024/09/30/2024.09.30.615220.full AB - We present a computational tool, MARLOWE, for source organism characterization of unknown, forensic biological samples. The intent of MARLOWE is to address a gap in applying proteomics data analysis to forensic applications. MARLOWE produces a list of potential source organisms given confident peptide tags derived from de novo peptide sequencing and a statistical approach to assign peptides to organisms in a probabilistic manner, based on a broad sequence database. In this way, the algorithm assumes no a priori knowledge of potential sources, and the probabilistic way peptides are taxonomically assigned and then scored enables results to be unbiased (within the constraints of the sequence database). In a proof-of-concept study, we examined MARLOWE’s performance on two datasets, the biodiversity dataset and the Bacillus cereus superspecies dataset. Not only did MARLOWE demonstrate successful characterization to true contributors in single source and binary mixtures in the biodiversity dataset, but also provided sufficient specificity to distinguish species within a bacterial superspecies group. These results suggest that MARLOWE is suitable for candidate- or lead-generation identification of single-organism and binary samples that can generate forensic leads and aid in selecting appropriate follow-on analyses in a forensic context.Competing Interest StatementThe authors have declared no competing interest.