CyVera Corporation proposes to develop and validate the feasibility of a rapid, robust, and inexpensive method for performing multiplexed protein expression measurements. These measurements are needed for the early detection, diagnosis, and the management of patients with cancer. This cancer diagnostic platform will be based on the combination of (i) CyVera's newly developed holograpically encoded, multiplexed microparticle assays, (ii) self assembly, and (iii) microfluidics. The format we propose will allow rapid and highly sensitive detection of protein expression patterns in small sample volumes, and will ultimately lead to a high-throughput instrument platform for cancer diagnostics. In Phase I of this project, prototype microfluidic devices will be constructed with antibody functionalized particles. Batches of individually encoded glass particles will be antibody functionalized, pooled, and self-assembled into microfluidic devices. Once assembled, the identity of each type of particle will be read via its holographic code. Five detection analytes in Phase I will be chosen from a set of putative cancer biomarkers. These commercially available markers will include von Willebrand factor (vWF), C-reactive protein (CRP), albumin, free Prostate Specific Antigen (fPSA), and complexed PSA (cPSA), all of which have been reported as prostate cancer biomarkers in the literature. The limit of detection and repeatability of each analyte will be assessed via spike-in experiments in serum samples. The goals of Phase I will be (1) to demonstrate < 10 pg/mL sensitivity of each multiplexed analyte in a complex sample in under one hour (2) low sample volume requirements of < 10 microliters, and (3) ease of fabrication and replication of the microfluidic devices. Success in Phase I will pave the way for the development of an affordable tool for molecular cancer diagnostics and follow-up patient therapy monitoring