The Next Generation of
Protein HOS Analysis

Reinventing Protein Higher Order Structural Analysis via State-of-the-Art Protein Footprinting

The Fox® Protein Footprinting System employs hydroxyl radicals (•OH) to examine the solvent accessibility of amino acid side chains within a protein. By selectively abstracting hydrogen atoms from exposed amino acids, •OH radicals induce the formation of oxidation products, enabling the detection and analysis of vital information concerning protein structure and dynamics. Automated HRPF, made possible by the Fox System, delivers crucial HOS information that greatly influences the safety, efficacy, and quality of biologics.

Comprised of a user-friendly benchtop instrument coupled with intuitive data analysis software, the Fox System makes it possible for biopharmaceutical researchers to easily and efficiently unravel the mysteries of protein HOS. This powerful tool opens new avenues for studying and understanding proteins, accelerating advancements in biopharmaceutical research and development.

Advantages

  • Direct Assessment of Solvent Accessibility

  • Higher-Order Structure Validation

  • High Spatial Resolution

  • Versatility Across Protein Classes

  • Complementary to Other Techniques

Insights

  • Protein-Protein Interactions

  • Epitope & Paratope Mapping

  • Protein-Ligand Interactions

  • Protein Folding & Conformational Changes

  • Regions Impacted by Allostery

  • Glycoprotein Structure & Function

  • Rapidly Test Structural Models

Hands-Free, High-Throughput
Hydroxyl Radical Protein Footprinting

With the launch of our new AutoFox® Protein Footprinting System, GenNext is empowering structural biology researchers to perform fully-automated Hydroxyl Radical Protein Footprinting (HRPF) without the need for expensive, complicated, and hazardous lasers. Plus, GenNext’s FoxWare® Software removes data processing bottlenecks with this powerful program tailored to meet the demands of HOS analysis.

Learn how our products can easily fit into your lab’s workflow or test-drive the Fox System on an outsourced project basis.

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HRPF Services for Monoclonal Antibodies

Let the experienced GenNext research team generate high-value information on the characterization of monoclonal antibodies (mAb) for more effective and safe biologics. Start a project today to study antigen-binding sites, epitope/paratope mapping, host antibody responses, antibody aggregation, and more.

In mAb design, use the high-resolution structural information generated from HRPF for epitope and paratope mapping to accelerate the rational design of therapeutics. In mAb production, use HRPF to monitor mAb structure during scale-up to ensure batch-to-batch structure stability over time, ensuring product shelf life and continued efficacy.

Unlike other protein structure analysis techniques, the Fox Protein Footprinting System delivers dynamic, in-solution structure and interaction information for a wide range of protein sizes, states, and concentrations.

Explore the Advantages of HRPF Via Fox Services

Complement your traditional HOS methods or begin a totally new route to modern HOS analysis. Check out our TOOLKIT to learn how our service delivers new and actionable data beyond traditional HOS methods.

GenNext News

Breaking News
GenNext Technologies and Thermo Fisher Scientific Collaborate to Simplify Structural Biology for Faster Biotherapeutic Development
GenNext and Agilent Unveil Groundbreaking Technology Integration to Accelerate High-Value Decision Making
Evaluating MS-Based HRPF of a Benchtop Fox System against a Synchrotron X-ray Beamline
GenNext Unveils the First Fully Automated HRPF System at the CASSS MS Meeting
GenNext & SCIEX to Co-Market an Advanced HOS Workflow for Biopharmaceutical Research
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GenNext & Thermo Fisher Collaborate to Simplify Structural Biology

New workflow consisting of GenNext's AutoFox System and Thermo Fisher's Orbitrap platform streamlines protein structural analysis and supports the growing role of AI in drug discovery.

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GenNext and Agilent Unveil Groundbreaking Technology Integration

Advancing structural biology research with the GenNext AutoFox® Protein Footprinting System and Agilent 6560 Ion Mobility Quadrupole Time-of-Flight (IM-QTOF) LC/MS system.

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GenNext & SCIEX to Co-Market an Advanced HOS Workflow

Together, the companies will spread the adoption of the latest HOS and Mass Spec technologies to help scientists improve the safety, stability, and effectiveness of biotherapeutics.

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GenNext Unveils the First Fully Automated HRPF System at CASSS MS

The new AutoFox System promises to deliver hands-free, high-throughput Hydroxyl Radical Protein Footprinting for sophisticated Higher Order Structural (HOS) analysis.

Featured On-Demand Scientific Presentations

The GenNext AutoFox® Protein Footprinting System combined with mass spectrometry enables amino acid-level mapping of protein interactions and conformational changes. Advanced MS tools like the Sciex ZenoTOF 7600 and Agilent 6560 address challenges include fragment ion identification for accurate oxidation localization and isobaric peptide co-elution causing chimeric spectra. Furthermore, integrating Thermo’s XL-MS further resolves ambiguous signals, advancing biologics and biotherapeutics research.

Presented by Dr. Emily Chea, Applied Research Manager of GenNext Technologies, Inc.

The AutoFox® Protein Footprinting System is a mass spectrometry-enabled platform for mapping protein structures and detecting interaction sites by oxidizing solvent-accessible amino acids. As structural changes or protein interactions affect solvent accessibility, modification levels shift accordingly. The AutoFox System is coupled with bottom-up proteomics and reverse-phase chromatography, to localize and quantify the extent of oxidation. Ion mobility separation further enhances resolution by resolving co-eluted, isobaric peptides before MS/MS fragmentation, improving residue-level precision.

Presented by Dr. Emily Chea, Applied Research Manager of GenNext Technologies, Inc

Many structural biology researchers seek easier and more robust methods for Higher Order Structural (HOS) analysis, recognizing its critical role in biotherapeutic stability, safety, and function. Traditional HOS approaches can be complex, slow, dangerous, and costly, or unreliable and uninformative. With many factors to consider, choosing the best method for your lab can be challenging. Dr. Chea reviews various options, highlighting their pros and cons, and introduces the Flash Oxidation (Fox®) Protein Footprinting System. She explains its applications in epitope mapping, mAb aggregation, and conformational analysis, helping you select the best method for developing safe and effective biotherapeutics.

Presented by Dr. Emily Chea, Applied Research Manager of GenNext Technologies, Inc.

Dr. Sharp discusses a covalent labeling technique called fast photochemical oxidation of proteins (FPOP), detailing the workflow, its application in research, and how a new platform is designed to enhance user-friendliness. The Fox Oxidation System streamlines the entire process, from radical generation to sample collection, improving safety, reliability, and ease of use compared to traditional FPOP setups. This innovation makes HRPF accessible to more researchers, allowing them to gain valuable insights into protein structures and interactions with greater ease.

Presented by Dr. Joshua Sharp, Research Institute of Pharmaceutical Sciences at the University of Mississippi.

This hour-long seminar, hosted with The Analytical Scientist, reviews structural biology case studies showcasing the successful application of protein footprinting in solving key problems in biopharmaceutical research. Examples include epitope and paratope mapping, protein interactions, receptor-drug interactions, and protein folding/unfolding. The seminar highlights the benefits of using protein footprinting technology in advancing biopharmaceutical research.

Presented by Professor Michael Gross of the Washington University in St. Louis.

A study on TNF alpha and its monoclonal antibody adalimumab, using the Fox Protein Footprinting System, revealed oxidation changes in TNF alpha peptides matching the known epitope. Adalimumab’s Fab region showed decreased oxidation due to reduced solvent accessibility when bound to TNF alpha, while the Fc region had increased oxidation, indicating flexibility beyond the crystal structure. This method accurately mapped the TNF alpha epitope and adalimumab paratope, providing insights beyond the crystal structure.

Presented by Dr. Emily Chea, Applied Research Manager of GenNext Technologies, Inc.

Using the Fox® System, researchers have advanced the understanding of therapeutic HOS by exploring epitope/paratope mapping, aggregation-interface identification, formulation, and small molecule binding. The Fox System, a novel HRPF technique, employs hydroxyl radicals (•OH) to irreversibly modify solvent-exposed amino acid side chains, revealing changes in solvent accessibility and uncovering buried or exposed regions. This presentation will discuss the Fox System, its application in characterizing biotherapeutic HOS and future innovations.

Presented by Dr. Emily Chea, Applied Research Manager of GenNext Technologies, Inc.

Mark Chance discussed his vision for proteomics and structural biology, focusing on megadalton protein complexes and the technologies needed to study them. He highlighted the importance of combining techniques like NMR, crystallography, cryo-EM, and mass spectrometry for protein interaction insights. He explained hydroxyl radical footprinting, a method for mapping protein surfaces and how it has advanced drug development. He shared examples from Rodeo Therapeutics, CASMA, and Foghorn Therapeutics, showcasing its practical applications.

Presented by Professor Mark Chance, Case Western Reserve University.

This 60-minute webinar, in collaboration with Cambridge Healthtech Institute, offers a comprehensive review and instruction in protein footprinting techniques and applications. Learn about various benchtop covalent modification methods and rapid labeling techniques like X-ray synchrotron and Fast Photo-Oxidation of Protein (FPOP) hydroxyl radical protein footprinting (HRPF). Explore their practical implications for biopharmaceutical research, protein interactions, biosimilar development, epitope and paratope mapping, ligand-binding, and conformational change analysis.

Presented by Professor Mark Chance of Case Western Reserve University and Professor Joshua Sharp of the University of Mississippi

Why Partner with GenNext?

Hear from luminaries in structural biology research on why they decided to join the GenNext Technologies Scientific Advisory Board to help develop and advocate for Flash Oxidation (Fox®) Hydroxyl Radical Protein Footprinting technology. These important partnerships underscore our commitment to advancing structural biology research and increasing the value of Fox technology in the biopharmaceutical industry.

Applications

The application of Fox-based HRPF technology in biopharmaceutical research is always expanding, from drug-binding, mAb therapeutics, and protein aggregation to formulation studies.

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Presentations

Watch recorded presentations, protocols, and interviews with GenNext scientists and our Scientific Advisory Board on on subjects ranging from applications to methods.

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HOS Technology

GenNext sifted through the available HOS analysis options to help you determine the most robust, cost-effective, and workflow-friendly approach for your lab.

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The Team

The GenNext team is an accomplished collection experts who have been creating first-in-class life science tools for both startups and industry giants.

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Publications

Learn about Hydroxyl Radical Protein Footprinting technology by reading the growing list of publications authored by GenNext scientists and members of our Scientific Advisory Board.

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Don’t Miss our 5-Part Protein Footprinting Seminar Series!

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Request a custom HRPF webinar for your lab.

Discover the Benefits of HRPF

Accelerate biopharmaceutical and biosimilar development and protein conformational research.

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