Technologies for Higher Order Structural Analysis

Many structural biology researchers are looking for a way to make Higher Order Structural Analysis easier and more robust because they know how crucial a role it plays in biotherapuetic stability, safety, and biological function. Yet, traditional HOS approaches are known to be either complex, slow, dangerous, and expensive or unreliable, uninformative, and irreproducible. Choosing the best approach for your lab can be confusing with so many factors to consider.

Dr. Chea takes you through your options, explaining the pros and cons of each method. Then she will provide an overview of the Flash Oxidation (Fox™) Protein Footprinting System and how it is being applied in epitope mapping, mAb aggregation, and conformational analysis. By better understanding your options for higher order structure analysis, you can choose the method that empowers your lab to develop the safest and most effective biotherapeutics.

Presented by Emily Chea, PhD

Dr. Emily Chea received her PhD while studying with Professor Dr. Lisa Jones at the University of Maryland, Baltimore. Emily has made significant contributions to the development of HRPF, with particular emphasis on in-cell FPOP (IC-FPOP) for proteome-wide structural biology characterization of cellular drug–target interactions. Currently, Emily is a key member of the GenNext scientific team in her role as an Applied Research Scientist.

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Meet Dr. Joshua Sharp

 Watch as Genny the GenNext Fox interviews the world-renowned and internationally recognized expert in Hydroxyl Radical Protein Footprinting (HRPF)—Dr. Joshua Sharp. Inventor of the first benchtop HRPF system and the pioneer of UV photolysis of hydrogen peroxide for hydroxyl radical generation, Josh has a lot of interesting things to say about HRPF.

JoVE Protocol

Online method to perform flash oxidation protein footprinting using inline radical dosimetry and a plasma light source.

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FASEB’s Protein Aggregation Conference:
Function, Dysfunction, and Disease

Flash Oxidation (Fox™) Hydroxyl Radical Protein Footprinting for mAb Aggregation

Spend less than 10-minutes with Dr. Chea as she explains how the Flash Oxidation (Fox™) Protein Footprinting System can characterize monoclonal antibody (mAb) aggregation and determine how excipients help stabilize the protein. Excipients, like amino acids, can stabilize proteins and limit aggregation. By comparing the change in hydroxyl radical modification before and after the addition of an amino acid, you can detect intermolecular interactions that take place during mAb aggregation. By better understanding the higher order structure of protein aggregation, you will be armed with the information you need to optimize mAb formulation and development.

Presented by Emily Chea, PhD

Dr. Emily Chea received her PhD while studying with Professor Dr. Lisa Jones at the University of Maryland, Baltimore. Emily has made significant contributions to the development of HRPF, with particular emphasis on in-cell FPOP (IC-FPOP) for proteome-wide structural biology characterization of cellular drug–target interactions. Currently, Emily is a key member of the GenNext scientific team in her role as an Applied Research Scientist.

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AAPS e-Chalk Talk

Revolutionizing Protein Higher Order Structural Analysis with Flash Oxidation (Fox™) Protein Footprinting

Learn about fundamentals of the HRPF along with applications relevant to the pharmaceutical sciences. Understand the  foundations of hydroxyl radical protein labeling chemistry and the relationship of labeling to protein topography. Fox technology, the methods used to generate radicals, 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 biopharmaceutical applications, while showing data highlighting higher order structure effects of formulations. Dr. Sharp will discuss 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.

Presented by Joshua Sharp, PhD

Dr. Joshua Sharp is an internationally recognized expert in HRPF, pioneering novel MS-based technology to study HOS focusing on the structure-function relationships of proteins and carbohydrates. With more than 21 peer-reviewed articles, Dr. Sharp has also presented numerous invited lectures around the world. Along with acting as the Chief Technology Officer of GenNext, Dr. Sharp is also Associate Professor of Pharmacology, Associate Professor of Chemistry and Biochemistry, and Director of the Glycoscience Center of Research Excellence at the University of Mississippi.

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ABRF 2021

The Flash Oxidation (Fox™) System: A Novel Laser-Free Photochemical Oxidation Protein Footprinting Platform

Dr. Joshua Sharp, Chief Technology Officer of GenNext Technologies and Associate Professor of Pharmacology, Associate Professor of Chemistry and Biochemistry, and Director of the Glycoscience Center of Research Excellence at the University of Mississippi, presents during the ABRF 2021 Virtual Meeting on the Fox Footprinting System.

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Presented by Joshua Sharp, PhD

Dr. Joshua Sharp is an internationally recognized expert in HRPF, pioneering novel MS-based technology to study HOS focusing on the structure-function relationships of proteins and carbohydrates. With more than 21 peer-reviewed articles, Dr. Sharp has also presented numerous invited lectures around the world. Along with acting as the Chief Technology Officer of GenNext, Dr. Sharp is also Associate Professor of Pharmacology, Associate Professor of Chemistry and Biochemistry, and Director of the Glycoscience Center of Research Excellence at the University of Mississippi.

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