ADVANCED DIAGNOSTICS USING RNA/PROTEIN FUSION TECHNOLOGY

The objective of this project is to develop advanced diagnostic and therapeutic monitoring systems for agents involved in allergy and inflammation. A key initiative in NIAID programs for Healthy People 2010 is the development of tools for drug discovery and monitoring allergy and inflammation that are associated with infectious disease and chronic disabling conditions. Cytokines and chemokines are mediators (or surrogate markers) of these disease processes. In this program, a novel immunoglobulin-like scaffold has been developed as an antibody mimic. From a large random library consisting of 10313-10e14 distinct members, binding proteins were rapidly selected in vitro against key mediators by inflammatory or allergic processes. Antibody mimics to a minimum of 5 cytokine or chemokine targets were selected and will be further evaluated for performance characteristics such as affinity, specificity, epitope mapping, and ability to function on a solid phase surface. If needed, binding proteins can be improved by affinity maturation, and PCR mutagenesis. The resulting antibody-mimics will be used for the development of a prototype addressable protein chip for detection of these markers in a variety of experimental samples. This multiplex microarray will be validated for the detection and quantification of cytokines and/or chemokines in culture supernatants, biological fluids, cells, or tissues. PROPOSED COMMERCIAL APPLICATIONS: This technology provides a powerful alternative to monoclonal antibodies and phage display mediated production of antibodies or fragments thereof. These unique protein binders will be incorporated into an addressable multiplex array for the high throughput analysis of samples from culture supernatants, biological fluids or cells for drug discovery and diagnostic applications.