Research Appraisalother

Protein Biomarkers as Molecular Tools for Forensic Biofluid Identification

Journal of proteome researchChhikara, Anjali, Gupta, Vinayak, Kumari, Pallavi et al.3 Apr 2026DOI

Clinical Snapshot

20CEBM
Evidence: Weakother

PICO Framework

P — PopulationForensic biological samples (blood, saliva, semen, tissues)
I — InterventionProteomic analysis using mass spectrometry and bioinformatics
C — ComparatorTraditional DNA analysis and conventional forensic methods
O — OutcomesBiofluid identification accuracy, post-mortem interval estimation, species identification

Bottom Line

This narrative review examines the potential of proteomic biomarkers in forensic applications, particularly for biofluid identification and post-mortem interval estimation. While the technology shows promise for enhancing forensic investigations when DNA analysis is insufficient, the review lacks systematic methodology and provides no quantitative evidence synthesis. The authors appropriately acknowledge significant barriers to implementation, including equipment costs, sample stability issues, and the need for standardized protocols that meet legal standards. The field remains in early development, requiring extensive validation studies before routine forensic application. Healthcare professionals should view this as an emerging technology with potential future applications rather than a ready-to-implement forensic tool.

Evidence: Weak

Key Findings

  • Effect Size: Not quantified

  • Primary Outcome: Forensic proteomics shows promise for biofluid identification and PMI estimation

  • Nnt Or Sensitivity: Sensitivity and specificity values not reported

  • Confidence Interval: Not provided

Clinical Application

Currently limited by high costs, equipment requirements, and lack of standardization Would require validation under Australian forensic standards and potential integration with existing forensic protocols Forensic laboratories and legal medicine practitioners

Abstract

Forensic proteomics has rapidly established a significant role in forensics, particularly when DNA analysis is insufficient. Proteomics has been frequently recognized for helping reveal useful information about the origin, state and context of forensic samples. Proteins are considered crucial biomarkers in biological samples because of their ability to elucidate cellular functions and post-mortem alterations. Combining advanced mass spectrometry and bioinformatics has increased the importance of proteomics in forensic sciences. The current study examined the use of proteomic technologies, mass spectrometry, and sophisticated software across the key domains of forensic research. The procedure for Post-Mortem Interval (PMI) estimation and body fluid classification proved to be more precise because of the use of machine learning. Peptide biomarkers have helped identify various species by samples of blood, saliva, and semen, and recognize brain, muscles, and skin tissues. Despite significant advancements, the wide acceptance of forensic proteomics remains problematic due to intricate sample stability, high equipment costs, and strict legal standards. Recent advancements in analytical sensitivity, data interpretation tools, and collaborative efforts toward robust protocols position forensic proteomics as an indispensable component of the forensic toolkit. This review indicates the increasing relevance of proteomics in forensic applications, especially relating to PMI estimation, body fluid differentiation, and disease profiling. It promises to significantly enhance the depth of evidentiary interpretation and contribute to more precise and equitable outcomes in the criminal justice system. Proteomic biomarkers need further validation across a range of environments, and standardized protocols should be developed and tested to ensure that proteomics is suitable for forensic use in the courts.

References

  1. 1.Chhikara, A., Gupta, V., Kumari, P., & Dalal, J. (2026). Protein biomarkers as molecular tools for forensic biofluid identification. Journal of Proteome Research. https://doi.org/10.1021/acs.jproteome.5c00898
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