Recombinant Proteome Expression and Antibody Selection

79636S An important goal of the Genomes-to-Life program is the development of comprehensive databases of structural and functional information on the proteomes of organisms of interest to DOE. This effort will require high-throughput methods for (1) mapping all pair-wise and higher-order interactions among the members of each such proteome, (2) optimizing the heterologous expression of proteome members for interaction mapping and also for purification and analysis in vitro, and (3) obtaining high-affinity reagents for proteome members. In this project, a high-throughput system will be developed for optimization of heterologous protein expression in prokaryotic or eukaryotic cells, utilizing a selectable protein-protein interaction sensor and peptide chaperones. In Phase I, the heterologous protein expression optimization system will be constructed and tested using a model proteome. The expression efficiency of the proteome will be determined before and after selection with the system, and optimization of the system performance will be carried out. Finally, optimized proteome members will be challenged with an antibody library to demonstrate high-throughput antibody selection. Commercial Applications and Other Benefits as described by the awardee: The technology should enable the high-throughput characterization of the structures and functions of all gene products of organisms of interest. These tools would be of use to the many companies engaged in mining human, animal, plant, and micro-organismal genomes for pharmaceutical and industrial applications. Future products could include new targets for the diagnosis and treatment of human, animal, and agricultural diseases; new and better assays for environmental pollutants and biological and chemical weapons; genetically engineered organisms for environmental and industrial applications in bioremediation and bio-organic synthesis and production; and new assays for the discovery and toxicological evaluation of new drugs and other bioactive molecules.