Protein markers for use in forensic and biometric applications

A survey of over 15 years' worth of relevant forensic and biomedical literature reveals a shortlist of candidate protein markers for use in protein-based forensic analysis. The selection (currently based on over 200 publications, not listed here due to size limitation) is summarized in Table 13.1. For clarity we have divided these into five main categories: Biometrics, Blood Origin, Lifestyle, Time of Death, Trauma and Death. We have also included a few 'small molecule' metabolites (e.g. testosterone, etc.) as these are also relevant to molecular foren-sics and antibodies are commercially available and the same assay format (CDA; see Barry et al., 2003; Barry and Soloviev, 2004) can be used. We have recently obtained 40 antisera against 20 of the -100 targets listed in Table 13.1. Prediction of antigenic peptide sequences is a crucial part of any antibody generation programme; the chosen markers and their respective anti-peptide antibodies are shown in Table 13.2 (for abbreviated protein names, see the legend to Table 13.1). The choice of the peptides was made on the basis of their predicted immunogenicity using in-house software (manuscript in preparation).

Another parameter requiring special attention is the choice of the solid support for the microarrays. Extensive data on protein array production parameters have been published in recent years by us and others (Kuznezow et al., 2003; Soloviev and Terret, 2005). Protein arrays have been produced on plastic or glass slides (most similar to DNA microarrays), hydrogels, filters or nitrocellulose (CASTĀ® and FASTĀ® Slides from Whatman/Schleicher Schnell), in microtitre plate wells or on beads. We have tested a number of surfaces to ensure maximum compatibility with crude rabbit antisera, the printing robot and the assay (in protein markers for use in forensic and biometric applications 209 Table 13.1 Protein markers for use in forensic and biometric applications

(a) Time of Death

Seasonal

Winter

Autumn/winter

Seasonal affective disorder

Diurnal

Sleep-wake behaviour

Sleep-wake behaviour

Time since death

Postmortem interval (PMI) marker

PMI up to 24-h marker

PMI (0-5 days) marker

Survival time

<5 min survival time (incised wounds) Neck injury: few min survival time Neck injury: several min survival time Neck injury: long time

(b) Lifestyle

Parallels IL-1, TNF

Cleavage of C3

Hypoxia-inducible levels of VEGF Degradation of cTnI ~ log(time)

Diet

Carbohydrate, polyunsaturated fat, animal protein, milk, dairy products, fish, poultry, minerals (K, Zn, Mg, Ca, P) High vegetable consumption, particularly tomatoes Western diet (red and processed meats, sweets, desserts, French fries, refined grains)

Prudent diet (higher intake of fruit, vegetables, legumes, fish, poultry, whole grains) Mediterranean diet Coffee consumption Moderate and severe malnutrition Protein malnutrition Diet, weight loss Alcohol consumption Alcohol intake (in healthy individuals and alcoholic liver disease) Chronic alcohol abuse Alcoholics Alcohol abstinence Smoking

Exposure to either cadmium or tobacco smoke

Smoking, intensive smoking Smoking cessation

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