A Microfluidic System for Affinity Reagent Generation and Assay Development for High-Throughput Biomarker Validation

Protein biomarkers are finding increasing application in Force Health Protection and health surveillance. However, very few biomarkers have been verified and validated. This deficiency has been ascribed primarily to availability of reagents (affinity probes). There is a need for assays to measure biomarker candidates, which must be accompanied by the development of reagents for detection. We propose to develop a microfluidic high-throughput reagent generation platform using capillary electrophoresis (CE) to enable the discovery and directed evolution of molecular probes from combinatorial libraries using an in vitro protein selection technique. The platform will be adapted to develop immunoassays using the probes selected. Key benefits include reduced reagent consumption (cost savings), automation, thermostable probes, and faster screening. In Phase I, the microfluidic system will be designed and fabricated, and proof-of-concept established by accomplishing probe screening and assay development for multiple protein biomarkers. In Phase II, the microfluidic system will be used to develop reagents for biomarkers provided by USACEHR. Currently, no miniaturized system to develop reagents in a cost-effective, high-throughput format is commercially available. A multidisciplinary team with proven expertise in microfluidics, protein engineering, and device development has been assembled.