Biomarkers are urgently needed for the early and accurate diagnosis of Alzheimer's disease (AD). SurroMed is using a high throughput, multiplexed proteomics approach to detect, identify and quantify a large number of putative biomarkers in animal models and human cerebrospinal fluid (CSF) specimens. Aim I proposes to develop methods to fractionate CSF and identify peptides and proteins in CSF using micro-scale liquid chromatography and mass spectrometry (MS). We will use proprietary bioinformatics to identify and quantify metabolites, peptides, proteins in mass spectra. Aim II proposes to identify specific candidate biomarkers related to underlying pathophysiological processes with known or hypothesized involvement in AD, specifically the ubiquitin proteasome system (UPS). We will develop conditions to simulate AD using proteasome inhibition in rodent primary neuronal cell cultures since proteasome inhibition is suspected to occur as an early event in AD pathology. We will use a variety of affinity techniques to capture peptides or proteins that accumulate due to UPS inhibition. Captured proteins will be identified by MS, and added to a list of putative biomarkers. Taken together, these studies will demonstrate the feasibility of our broad phenotypic analysis approach to identifying and quantifying biomarkers and provide us with validated methods and tools for studies of human CSF samples. PROPOSED COMMERCIAL APPLICATION: We propose to develop a global solution for proteomic analysis using an innovative technology platform for probing the molecular signatures of Alzheimer's disease (AD). The analysis will be coupled with proprietary bioinformatics to identify and validate putative biomarkers for AD. This research program will lead to commercialization of diagnostic kits, therapeutic targets, instrumentation, and bioinformatics.