Based on recent progress in sequencing the human genome, a new goal has emerged: to express and analyze the human proteome. The development of proteome arrays will accelerate progress in several key areas of biology and medicine including mapping cellular pathways, discovering new drug targets and identifying disease related biomarkers. However, the production of proteome arrays is currently hindered by the need for improved methods of expressing proteins from gene libraries, engineering proteins for site-specific surface attachment and detecting multiple protein-protein interactions. This project aims to overcome these limitations by combining cell-free methods for rapid expression of specially engineered protein libraries with advanced technology for microarray fabrication and readout.AmberGen will produce and evaluate proteome arrays constructed from cell-free expressed protein libraries and designed for high sensitivity and multiplex readout. The rapid detection of protein-protein interactions will be used to map cellular pathways and to discover new biomarkers. Prototype proteome arrays from the pathogen H. pylori and selected genes from the human genome will be constructed and evaluated in Phase I. Phase II will focus on applications in the areas of autoimmune and infectious diseases.