DNA profiling has been called the “gold standard” of modern forensic testing. The characterization and confirmatory identification of the tissue source of a DNA profile, however, still presents numerous challenges for forensic serologists. Current chemical reaction-, enzyme activity- and antibody-based assays used by forensic practitioners for body fluid identification yield only presumptive results. Positive results with non-target body fluids or cross-reactivity with non-human sources has been well documented. Some serological tests can consume unacceptable quantities of precious evidence while for other body fluids (e.g., vaginal fluids) there are no commercially available tests at all. Comprehensive proteome mapping and comparative multidimensional HPLC analyses in combination with Q-TOF mass spectrometry has resulted in the identification and verification of a series of protein “biomarker panels” for six forensically-relevant body fluids (i.e., peripheral and menstrual blood, vaginal secretions, semen, urine and saliva). These high-specificity panels have served as the foundation for the development of a multiplexed serological assay for the single-pass identification of these body fluids. Using the Agilent Technologies Agilent® AssayMAP Bravo automation platform for front end sample preparation and an Agilent 6495 triple quadrupole mass spectrometer coupled to a 1290 UHPLC for mass analysis, an efficient workflow has been developed to meet the demanding reliability and throughput needs of the forensic operational environment. To date, a 23-protein, 51-peptide panel for fluid identification has been developed with a run time of only 10 minutes. procedures and reliable interpretation guidelines. The data generated to date demonstrate the utility of mass In conjunction with the automation platform, several hundred samples can be analyzed per week. A series of developmental validation studies are currently in progress to establish performance limits of the assay (reproducibility, repeatability, sensitivity, case-samples, etc.) and to inform the creation of standard operating procedure for the use of spectrometry as a unified platform for high sensitivity confirmatory body fluid identification. This technology has the potential to replace the diversity of test methods currently employed with a simpler and more standardized work flow for forensic body fluid identification.

NextGen serology: Protein mass spectrometry for the forensic identification human body fluids

256th National Meeting (2018)

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