title: Verification of protein biomarker specificity for the identification of biological stains by quadrupole time-of-flight mass spectrometry.
abstract: Advances in proteomics technology over the past decade offer forensic serologists a greatly improved opportunity to accurately characterize the tissue source from which a DNA profile has been developed. Such information can provide critical context to evidence and can help to prioritize downstream DNA analyses. Previous proteome studies compiled panels of "candidate biomarkers" specific to each of five body fluids (i.e., peripheral blood, vaginal/menstrual fluid, seminal fluid, urine, and saliva). Here, a multiplex quadrupole time-of-flight mass spectrometry assay has been developed in order to verify the tissue/body fluid specificity the 23 protein biomarkers that comprise these panels and the consistency with which they can be detected across a sample population of 50 humans. Single-source samples of these human body fluids were accurately identified by the detection of one or more high-specificity biomarkers. Recovery of body fluid samples from a variety of substrates did not impede accurate characterization and, of the potential inhibitors assayed, only chewing tobacco juice appeared to preclude the identification of a target body fluid. Using a series of 2-component mixtures of human body fluids, the multiplex assay accurately identified both components in a single-pass. Only in the case of saliva and peripheral blood did matrix effects appear to impede the detection of salivary proteins.
authors.affiliation: Department of Biological Sciences, University of Denver, Denver, Colorado, USA.|The Center for Forensic Science Research and Education, Willow Grove, PA, USA.
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Legg
authors.affiliation: Department of Pharmaceutical Sciences, University of Colorado, Aurora, Colorado, USA.
authors.forename: Roger
authors.initials: R
authors.lastname: Powell
authors.affiliation: Department of Pharmaceutical Sciences, University of Colorado, Aurora, Colorado, USA.
authors.forename: Nichole
authors.initials: N
authors.lastname: Reisdorph
authors.affiliation: Department of Pharmaceutical Sciences, University of Colorado, Aurora, Colorado, USA.
authors.forename: Rick
authors.initials: R
authors.lastname: Reisdorph
authors.affiliation: Department of Biological Sciences, University of Denver, Denver, Colorado, USA.|The Center for Forensic Science Research and Education, Willow Grove, PA, USA.
authors.forename: Phillip B
authors.initials: PB
authors.lastname: Danielson
mesh_terms: D015415:Biomarkers / Q000737:chemistry; D001826:Body Fluids / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D005260:Female; D005554:Forensic Medicine; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D010455:Peptides / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D020543:Proteome / Q000032:analysis*; D012333:RNA, Messenger / Q000032:analysis
chemical_list: D015415:Biomarkers; D010455:Peptides; D011506:Proteins; D020543:Proteome; D012333:RNA, Messenger
keywords: Forensic biology; Forensic serology; Mass spectrometry; Proteomics