Author Identifier
Date of Award
2026
Keywords
high resolution mass spectrometry, proteomics, taxonomic classification, forensics
Document Type
Thesis
Publisher
Edith Cowan University
Degree Name
Doctor of Philosophy
School
School of Science
First Supervisor
Hayley Abbiss
Second Supervisor
Joel Gummer
Third Supervisor
Glendon Parker
Abstract
Hair fibres constitute a common and persistent form of trace biological evidence in forensic investigations. The determination of hair origin can provide valuable forensic intelligence and assist in the prioritisation of evidentiary material. Traditional classification methods based on hair morphology require specialised expertise, offer limited taxonomic resolution, and are not readily scalable. This thesis examines the application of mass spectrometry based proteomics as an alternative biomolecular approach for the taxonomic classification of hair, with emphasis on taxonomically informative biomarker discovery and translation into forensic workflows.
A discovery phase based on high resolution mass spectrometry was established to characterise hair proteomes from a diverse panel of human and non-human species of forensic relevance. A reproducible protocol for protein isolation and tryptic digestion from trace hair material was developed, enabling identification of 226 taxonomically informative peptides, of which 207 are newly reported. These markers were derived predominantly from keratins and keratin associated proteins and included peptides with both species-level and higher-order taxonomic discriminatory value. A curated hair protein sequence database and associated analytical workflow were subsequently developed to support classification of fur-hair samples without prior assumptions regarding species origin.
To facilitate implementation within existing forensic laboratories, the peptide biomarkers identified during the discovery phase were translated into a targeted analytical format using multiple reaction monitoring on triple quadrupole instrumentation. A qualitative assay for the identification of human hair was developed and validated, demonstrating reproducible detection of diagnostic peptides from sub-millimetre fragments. The targeted workflow was subsequently extended to non-human taxa, establishing the feasibility of streamlined peptide panels for broader forensic hair classification.
A hybrid biomarker discovery workflow was developed for native marsupial species, enabling identification of both species-specific and higher-order taxonomic markers despite sparse reference proteome coverage. To this end, addressing the limited representation of native Australian fauna in public protein sequence databases. Incorporation of these markers into the targeted liquid-chromatography mass spectrometry assay demonstrated utility for the triage and interpretation of environmentally derived mixed hair evidence.
Collectively, this thesis establishes a robust analytical framework for forensic hair proteomics spanning biomarker discovery, taxonomic classification, and targeted implementation, demonstrated the potential of proteomic analysis to extend the forensic value of hair evidence.
Access Note
Access to this thesis is embargoed until 4th July 2029
Recommended Citation
Keane, R. (2026). Characterisation of genetically variant proteins using high-resolution mass spectrometry for taxonomic assignment of biological evidence in the forensic setting. Edith Cowan University. https://doi.org/10.25958/kd9d-dn54