Document Title: Establishment of a Fast and Accurate Proteomic Method for Body Fluid/Cell Type Identification

Body fluid identification plays an important role in forensic investigations both for directing the course of further inquiries, as well as in contributing to the evidence in a case. Yet current methods for body fluid detection vary widely in sensitivity, reproducibility, speed, cost and above all specificity – i.e. their ability to confirm that test results are conclusive, in addition many tests are founded on techniques that are decades old and have failed to keep pace with recent advances in science and technology. Two examples are the routinely used tests for blood (Kastle-Meyer) and saliva (amylase). Both assays give false positives for a variety of naturally occurring compounds, both rely on the subjective evaluation of qualitative results based on an analyst’s best judgment, and both are more than 100 years old. Yet even the more quantitative and confirmatory immunoassays, like ELISA, suffer from the inherent limitations of antibody specificity (cross-reactivity), affinity (sensitivity), narrow working ranges (the Hook effect), and manufacturing production variability, all of which can contribute to varying results within and between testing laboratories. Additionally, there are many body fluids for which no tests are available, e.g. menstrual blood and vaginal fluid. The medley of body fluid tests with their significant variability in results (confirmatory vs. presumptive, different sensitivities, etc.) stands in contrast to DNA STR testing, which uses a single methodology on a wide variety of sample types (cells, body fluids, latent fingerprints, hair, etc.) to produce consistent results held to common accepted standards. Part of the advantage inherent of DNA testing is that it focuses on a single biopolymer – DNA, making it relative easy to hone extraction, purification, testing and analysis to a common method. Yet most confirmatory body fluid tests also rely on the detection of a single type of biopolymer – proteins. And although protein chemistry is more variable and complex than that of DNA (there are twenty different amino acids each with distinct chemical properties compared with only four nucleotides for DNA), scientific and technical advances in protein chemistry and analysis now bring it within range of common methods and standards for forensic body fluid identification. The overall objective of this project was to determine if a single, confirmatory methodology, employing mass spectrometry (MS) as the means of detection, could be developed for the