eChalk Talk: Revolutionizing Protein Higher Order Structural Analysis with Flash Oxidation (Fox) Protein Footprinting
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This chalk talk will offer an overview of Hydroxyl Radical Protein Footprinting, focusing on explaining the fundamentals of the technique and applications relevant to the pharmaceutical sciences. The foundations of hydroxyl radical protein labeling chemistry and the relationship of labeling to protein topography will be reviewed. Furthermore, a brief description of the Flash Oxidation (Fox) technology and the method used to generate radicals as well as the method for real-time dosimetry and the requirements for sample analysis by LC-MS/MS will be presented. Dr. Sharp will present examples of applications, focusing on the analysis of biopharmaceuticals, along with presenting data highlighting higher order structure effects of formulations. Dr. Sharp will show the effects of different buffers on the aggregation propensity and aggregation interfaces of a therapeutic monoclonal antibody along with verifying the ability of polysorbate to prevent aggregation without changing the topography of the monomeric protein. He will also present results defining the epitope of a therapeutic antibody. These applications illustrate a few of the potential applications of Fox technology towards the analysis of protein biopharmaceuticals.