HT-production of Abs using Interaction Trap in Yeast

DESCRIPTION (provided by applicant): Affomix would like to commercialize antibody chips to enable researchers to study the human proteome. Currently, researchers use gene chips as a surrogate means to estimate the concentration of the vast variety of pro teins in cells and tissues. But given the choice, most researchers would prefer to study the proteins directly because of the; i) poor correlation between gene expression and protein concentration, and ii) lack of any information regarding post-translatio nal modification when using gene chips. The problem has been that bench-level methods for studying protein levels and post-translational modification occurring in cells and tissues are not available. Antibody chips would address this problem but until now there has not been a cheap and facile method to isolate antibodies against the nearly 25,000 proteins in the human proteome. Our strategy is to create a new approach for the high-throughput (HT) isolation of high-affinity antibodies. The overall plan is to develop and use an automated format of the yeast two-hybrid (Y2H) system to provide a commercially viable molecular biology 'assembly line' by which to generate thousands of different antibodies. Affomix has three primary commercial obje ctives; 10 supply researchers with high-quality antibody chips, 2) perform custom services for the identification of human antibodies against specified targets, and, 3) develop our own antibody therapeutics. Successful completion of this Phase I proposal will allow us to complete these first 2 objectives. The significant short-term objective of this specific grant application is to establish a robust assay for estimating the strength of a given antibody:antigen interaction. Successful completion of the f our state specific aims will set the state for the increased efficiencies and analysis in a future scaled-up effort. Completion of our Phase II specific aims will allow us to provide researchers with significant new reagents with which to study the proteo mic makeup and diversity of cells, and facilitate efforts to identify, track, quantify, control, capture, and image both individual proteins and molecular mechanisms in living systems.