SBIR-STTR Award

The GenNext Phase I SBIR submssion entitled “Multi-Wavelength Fluorescence Radical Dosimetry for Real-Time Assessment of Protein Footprinting Radical Yield” is responsive to the ackowledged need for new and improved tools for higher order structural analysis (HOS) of biopharmaceuticals and membrane protein target studies. An emerging HOS analysis technique is hydroxyl radical foot-printing (HRPF). HRPF 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. We have developed commercial solutions to perform HRPF. Recently a new and valuable footprinting technique that relies upon trifluoromethyl (TFM) radicals created by OH radical attack of aqueous sodium triflinate has been developed that shows great promise when combined with HRPF. TFM protein footprinting (TFMPF) is highly complementary to HRPF, as TFMPF effectively labels amino acid residues that are relatively “silent” to OH radical attack. When used together, HRPF and TFMPF provide substantial coverage and detection of solvent accessible residues, and as such represent a transformative improvement to biopharmaceutical HOS assessment.The practice of TFMPF has been pioneered by Professor Michael Gross of Washington University, St. Louis, Mo. While showing great promise to address unmet challenges in pharmaceutical research, reproducibility for TFMPF is challenged by variability of background scavenging. Collaborating with the Gross laboratory, our work will extend our innovative HRPF radical dosimetry technology to TFMPF. GenNext Technologies is the only company commercializing products for HRPF HOS analysis. Our goal is to convert the combined use of HRPF and TFMPF process from an academic research experiment into a valuable analytical tool. Once simplified and transformed into a robust technique, we envision the facile combination of HRPF and TFMPF to enable: paratope and epitope the interaction of mAb biopharmaceuticals with their membrane targets; elucidate the dynamics of lead binding to orthosteric or allosteric membrane targets; to reveal secondary messenger signaling cascades of GPCR lead compounds; and to detect the impact of orthosteric / allosteric anti- neoplastics upon targets such as kinases and growth factors. Public Health Relevance Statement The importance of biopharmaceuticals and membrane protein targets has created a need for improved analytics to facilitate biopharmaceutical research. Our Phase I SBIR proposal will create an improved means to study the higher order structure of bipharmaceuticals and their intended druggable targets in the context of facilitating proteomics and pharmaceutical research and development. Upon successful completion of our program, we will demonstrate the transformative nature of our new technology to positively impact biopharmaceutical research.

Project Terms:
Adoption ; Amino Acids ; aminoacid ; Antibodies ; Monoclonal Antibodies ; Clinical Treatment Moab ; mAbs ; Epitopes ; Antigenic Determinants ; Binding Determinants ; Antineoplastic Agents ; Anti-Cancer Agents ; Antineoplastic Drugs ; Antineoplastics ; Cancer Drug ; Neoplastic Disease Chemotherapeutic Agents ; Tumor-Specific Treatment Agents ; anti-cancer drug ; anticancer agent ; anticancer drug ; Award ; Behavior ; Binding Sites ; Combining Site ; Reactive Site ; Antibody Binding Sites ; Paratopes ; antibody combining site ; Biological Products ; Biologic Products ; Biological Agent ; biopharmaceutical ; biotherapeutic agent ; Biological Response Modifier Therapy ; Biologic Therapy ; Biological Therapy ; biological therapeutic ; biological treatment ; biotherapeutics ; biotherapy ; Blood capillaries ; capillary ; Chemistry ; Comparative Study ; Dangerousness ; Pharmaceutical Preparations ; Drugs ; Medication ; Pharmaceutic Preparations ; drug/agent ; Environment ; Fluorescence ; Fluorine ; F element ; Goals ; Human ; Modern Man ; Industry ; Laboratories ; Lasers ; Laser Electromagnetic ; Laser Radiation ; Lead ; Pb element ; heavy metal Pb ; heavy metal lead ; Ligands ; Membrane Proteins ; Membrane Protein Gene ; Membrane-Associated Proteins ; Surface Proteins ; Molecular Conformation ; Molecular Configuration ; Molecular Stereochemistry ; conformation ; conformational state ; Patients ; Pharmacology ; Phosphotransferases ; Kinases ; Phosphotransferase Gene ; Transphosphorylases ; Play ; Production ; Protein Conformation ; Proteins ; Research ; research and development ; Development and Research ; R & D ; R&D ; Role ; social role ; Safety ; Signal Transduction ; Cell Communication and Signaling ; Cell Signaling ; Intracellular Communication and Signaling ; Signal Transduction Systems ; Signaling ; biological signal transduction ; Sodium ; Na element ; Solvents ; Technology ; Time ; Toxin ; Universities ; Washington ; Work ; Hydroxyl Radical ; Hydroxyl ; Generations ; base ; detector ; Label ; improved ; Surface ; Phase ; Link ; Biological Process ; Biological Function ; Letters ; Immunological response ; host response ; immune system response ; immunoresponse ; Immune response ; tool ; Nature ; programs ; Adopted ; Reaction ; Techniques ; interest ; membrane structure ; Membrane ; Performance ; Hydrophobicity ; professor ; dosimetry ; aqueous ; Structure ; novel ; G Protein-Complex Receptor ; G Protein-Coupled Receptor Genes ; GPCR ; G-Protein-Coupled Receptors ; novel technologies ; new technology ; Appearance ; Proteome ; Proteomics ; Protein Footprinting ; Molecular Interaction ; Binding ; Pharmaceutical Agent ; Pharmaceuticals ; Pharmacological Substance ; Pharmacologic Substance ; Extracellular Protein ; Address ; Detection ; Higher Order Chromatin Folding ; Higher Order Structure ; Higher Order Chromatin Structure ; NIGMS ; National Institute of General Medical Sciences ; Reproducibility ; Small Business Innovation Research Grant ; SBIR ; Small Business Innovation Research ; Transmembrane Domain ; TM Domain ; Transmembrane Region ; Monitor ; Process ; Modification ; pathogen ; innovation ; innovate ; innovative ; analytical tool ; Drug Targeting ; Growth Factor ; Growth Agents ; Growth Substances ; Proteins Growth Factors ; experimental study ; experiment ; experimental research ; druggable target ; pathogenic virus ; viral pathogen ; virus pathogen ; adverse drug reaction ;

The GenNext Phase II SBIR submssion entitled "Multi-radical Protein Footprinting (MRPF™) Platform," is responsive to the ackowledged need for new and improved tools for higher order structural analysis (HOS) of biopharmaceuticals and membrane protein target studies. An emerging HOS analysis technique is radical foot- printing (RPF). RPF involves the irreversible labeling of a protein's exterior by reaction with radicals with subsequent MS analysis to identify the outer portions of the protein. We have developed commercial solutions to perform Hydroxyl Radical Protein Footprinting (HRPF) employing OH radicals known as the Fox® Protein Footprinting System. A new RPF technique that relies upon trifluoromethyl (TFM) radicals created by "¢OH attack of aqueous sodium triflinate has been developed. TFM protein footprinting (TFMPF) is highly complementary to HRPF. When used together, HRPF and TFMPF provide improved spatial coverage than either alone. In our phase I program, we created a facile means to assess effective radical load during CF3 labeling experiments by creating an in-line fluorescence detector that measured the photometric response of coumarin-based internal standard radical dosimeters to "¢OH and "¢CF3 attack. In our Phase II program, we build upon our phase I success to create a pre-commercial system that will enable selectable "¢OH or simultaneous "¢OH and "¢CF3 labeling, with in-line radical dosimetry, and optimized data processing software. In doing so, we will extend our products to also support TFMPF, creating a powerful and impactful improvement to biopharmaceutical HOS. GenNext Technologies is the world's only company commercializing instruments and solutions for RPF HOS analysis. Our overarching goal is to convert the RPF process from an academic research experiment into a broadly adopted and valuable HOS analytical tool. We firmly believe that our proposal will significantly impact HOS analysis with transformative benefits to biopharmaceutical research.

Public Health Relevance Statement:
The importance of biopharmaceuticals and membrane protein targets has created a need for improved analytics to facilitate biopharmaceutical research. Our Phase II SBIR proposal will create a needed instrumentation means to study the higher order structure of membrane protein targets as well as other biopharmaceutical drug targets, facilitating proteomics and pharmaceutical research and development. Upon successful completion of our program, we will demonstrate the transformative nature of our new technology to positively impact biopharmaceutical research. Terms: