The overarching goal of Phase I is to produce a method that will generate, identify and clone a catalog of protein-protein interactions in a single rapid experiment. In this proposal we will model the technology using antibody-antigen interactions. Recombinant antibody libraries against a proteome could be produced prior to any need, and then desired affinity reagents systematically cloned as needed. Libraries of recombinational biopanning integration product against a proteome could also be distributed along with primer sets against a set of barcodes for facile cloning. In future endeavors, the method will be used to clone tumor-cell or other diseased tissue cDNAs to use as an epitope library to quickly identify potential biomarkers. Successful completion of this phase will allow th commercialization of antibodies that can be used for diagnostic and therapeutic applications at a speed and comprehensiveness that is not easily achieved by current methods. Kits composed of vectors, reagents, strains and methods would be of commercial interest.
To develop a technology that can screen a library of >1010 different antibodies against a library of >104 different protein (and/or peptide) antigens. The proposed technology should be applicable to any single- or mix of organisms, including viral, bacterial and human proteomes. An ability to quickly assemble affinity reagents for testing tumors and damaged organs, for example for diagnostic and biotherapeutics, is contemplated. Testing post-translationally modified proteins displayed on the cell surface using suppressor tRNAs will be discussed briefly.