Liquid Chromatography Flash Oxidation (LC-Fox™) Protein Footprinting System

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The GenNext Phase I SBIR submssion entitled “Liquid Chromatography Flash Oxidation (LC-Fox™) Protein Footprinting System,” is responsive to the ackowledged need for new and improved tools for higher order structural analysis (HOS) of biopharmaceuticals. Hydroxyl radical foot-printing (HRPF) is a HOS analytical technique that involves the irreversible labeling of a protein’s exterior by reaction with hydroxyl radicals with subsequent MS analysis to identify the outer portions of the protein. When applied to dynamically changing mixtures of conformation, HRPF fails to elucidate individual conformers. Our LC-Fox Protein Footprinting is a new, bench-top method to probe dynamic conformational heterogeneous mixtures that can be separated via various forms of chromatography, including size exclusion (SEC), ion exchange (IEX), and hydrophobic interaction chromatography (HIC). In Phase I we will create a prototype system to be tested with isocratic SEC. GenNext Technologies is the only company commercializing technology and products for HRPF HOS analysis. Our overarching goal is to convert the HRPF process from an academic research experiment into a broadly adopted and valuable HOS analytical tool. Our LC-Fox system is predicted to make substantial impact in the field of biopharmaceutical research / development, formulation, and drug delivery, as it will enable the detection and characterization of unwanted conformers (and potential adverse drug reactions) created during the genesis and introduction of new biotherapeutics.