SBIR-STTR Award

The objective of this proposal is to apply novel affinity mass spectrometry (AMS) technologies to identify protein biomarker fingerprints indicative of alcohol consumption. The methodology used combines micro-affinity protein capture from complex biological solutions with mass spectrometric detection. In this first phase (R43), we will screen a small number of plasma samples (n=48) to identify candidate protein profiles, that can appropriately differentiate sample origin from healthy abstinent controls from individuals differentially exposed to alcohol. These samples will be screened with affinity pipettes employing numerous wide specificity affinity surfaces, which include hydrophilic, hydrophobic and metal chelating ligands to name a few. Each of these ligand surfaces possesses specific affinity characteristics that will result in the capture and purification of various protein groups. Observed molecular profiles (fingerprints) that have statistically significant features that correctly group samples will be viewed as potential biomarkers. In the second (validation) phase (R44), we will construct immuno-targeted assays for identified significant features determined within each biomarker fingerprint and individually validate them against a larger number of samples (n=400). In the event that multiple protein biomarkers are validated, a multiplexed immuno-based AMS panel for alcohol exposure will be developed. This will be achieved through the construction of multi-analyte affinity pipettes that will selectively retrieve the targeted biomarkers that differentiate healthy from diseased states. Such panels will also be subject to the same validation process that each of its components individually endured. The ultimate result of this research will be a validated protein biomarker assay(s) that can be used to screen for alcohol exposure.

Thesaurus Terms:
alcoholic beverage consumption, biomarker, matrix assisted laser desorption ionization, method development chelating agent clinical research, human subject

The objective of this proposal is to validate potential biomarkers of excessive alcohol exposure discovered in the Phase 1 portion of this work (1R43AA016233-01). This will entail the development of pipettor tip based immuno-affinity mass spectrometry (AMS) devices for the routine and repetitive high throughput analysis of the biomarkers in question. Resultant devices will allow for both quantitative and qualitative differentiation of the markers of interest. With the successful development of immuno-AMS devices for the biomarkers in question, they will be used to screen clinical samples collected by our clinical collaborators (Aurora Psychiatric Hospital, Wauwatosa, WI) in order to validate their performance. Final metrics for the evaluation of the biomarker assays developed will be a clinical sensitivity and specificity of 85%, with coefficients of variation of <15%.

Public Health Relevance:
Alcohol abuse has a significant impact on our society, yet there are no accurate biomarker tests for prognostic, diagnostic or treatment efficacy assessment currently available. This application is to validate new biomarkers of excessive alcohol exposure using a novel in pipettor tip immuno-affinity mass spectrometry based assay. Clearly, accurate and specific biomolecular assays are of great importance, positively impacting the associated health, financial and emotional repercussions of alcohol abuse.

Thesaurus Terms:
Acute; Affinity; Alcohol Abuse; Alcohols; Antigens, Differentiation; Assay; Bioassay; Biologic Assays; Biological Assay; Chemical Class, Alcohol; Chronic; Clinical; Clinical Sensitivity; Development; Devices; Diagnostic Or Prognostic Tests; Differentiation Antigens; Differentiation Markers; Educational Workshop; Emotional; Engineering; Engineerings; Evaluation; Expression Profiling; Expression Signature; Feedback; Funding; Goals; Grant; Health; Heavy Drinking; Hospitals, Psychiatric; Marker Antigens; Markers, Differentation; Mass Spectrum; Mass Spectrum Analysis; Medicine; Mental Hospitals; Mental Institutions; Metric; Molecular Fingerprinting; Molecular Profiling; Niaaa; National Institute On Alcohol Abuse And Alcoholism; Nature; Performance; Phase; Photometry/Spectrum Analysis, Mass; Process; Programs (Pt); Programs [publication Type]; Proteins; Psychiatric Hospitals; Research; Sampling; Science Of Medicine; Sensitivity And Specificity; Societies; Spectrometry, Mass; Spectroscopy, Mass; Spectrum Analyses, Mass; Spectrum Analysis, Mass; Suggestion; Treatment Efficacy; Validation; Variant; Variation; Work; Workshop; Addiction; Alcohol Exposed; Alcohol Exposure; Alcohol Problem; Base; Biomarker; Cohort; Drink Heavily; Drinking; Ethanol Abuse; Ethanol Exposed; Ethanol Exposure; Excess Alcohol Consumption; Excess Alcohol Ingestion; Excess Ethanol Ingestion; Excessive Alcohol Consumption; Excessive Alcohol Ingestion; Excessive Alcohol Intake; Excessive Drinking; Excessive Ethanol Ingestion; Experiment; Experimental Research; Experimental Study; Exposed To Alcohol; Exposure To Alcohol; Extreme Drinking; Gene Product; Hazardous Alcohol Use; Heavy Alcohol Use; High Throughput Analysis; Interest; Meetings; Mental Health Facility; Molecuar Profile; Molecular Signature; Novel; Problem Drinking; Programs; Public Health Relevance; Research Study; Response; Therapeutic Efficacy; Therapeutically Effective