Applications and Methods for Continuous Monitoring of Physiological Chemistry

With the notable exception of glucose, most metabolic biomarkers cannot be accurately, and practically measured over long periods, in vivo and in real-time. A primary reason is stable, selective enzymes suitable for biosensors are not available. Biosensors could revolutionize acute and long-term monitoring in the development, clinical, first responder and battlefield arenas. Pinnacle has developed intellectual property (patent PCT/US11/51193) that provides a general solution for the use of cofactors to enhance the biosensors sensitivity. We have also discovered that fusion proteins can dramatically enhance the stability of oxidases without compromising catalytic activity. Using high throughput screening in a cell-free system, directed evolution and large scale protein production in E.coli, we propose to develop a platform methodology for the optimization of existing (e.g. histamine), and the creation of new, (e.g.cortisol) oxidases for biosensors. Pinnacle Technology manufactures, sells and warranties biosensors for the detection of glucose, glutamate, lactate, and alcohol in animals. With the Richter and Gao laboratories at the University of Kansas we have successfully cloned, expressed, purified and characterized oxidases for biosensors. The enzymes produced will be available for direct sale, or as biosensors. World-wide total biosensor sales are expected to reach $14 billion by 2016.