The goal of this project is to develop a high-throughput, standardized system for measuring large numbers of cellular or excreted proteins simultaneously. The key discovery to be exploited to accomplish this is in vitro RNA-protein fusion technology being pioneered at Phylos. In brief, this technology allows for the rapid identification, selection and engineering of proteins (including antibodies) with unique characteristics. As such, it provides a rapid alternative for the generation of diagnostic monoclonal antibodies. They now propose to establish methodology for selecting antibody mimics to multiple targets simultaneously. As proof-of-principle for the Phase I portion of this project, they propose to identify high-affinity binders to five cytokines (TNFalpha, IFNgamma, IL2, IL6, and IL12) selected to each target individually as well as in batch-type protocols to all five. The activity of antibody mimics identified by these two methodologies will be characterized and compared with the activity of commercially available monoclonal antibodies. The presence of RNA covalently fused to the encoded protein not only enables the selection and """"""""evolution"""""""" of high-affinity binders but also allows for a means to anchor these molecules via Watson-Crick base pairing onto DNA chips. The result is an addressable chip for detection of multiple specific proteins. Therefore, the ability of antibody mimic fusion molecules to detect their target in chip format will also be characterized.
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