Multi-Wavelength Fluorescence Radical Dosimetry for Real-Time Assessment of Protein Footprinting Radical Yield

用于实时评估蛋白质足迹自由基产量的多波长荧光自由基剂量测定

基本信息

批准号：
10250755
负责人：
Scot Randy Weinberger
金额：
$ 25.14万
依托单位：
GENNEXT TECHNOLOGIES, INC.
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-01 至 2023-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10250755
关键词：
Address Adopted Adoption Amino Acids Antibodies Antibody Binding Sites Antineoplastic Agents Appearance Award Behavior Binding Binding Sites Biological Process Biological Products Biological Response Modifier Therapy Blood capillaries Chemistry Comparative Study Dangerousness Detection Drug Targeting Environment Epitopes Extracellular Protein Fluorescence Fluorine G-Protein-Coupled Receptors Generations Goals Growth Factor Higher Order Chromatin Structure Human Hydrophobicity Hydroxyl Radical Immune response Industry Label Laboratories Lasers Lead Letters Ligands Link Membrane Membrane Proteins Modification Molecular Conformation Monitor Monoclonal Antibodies National Institute of General Medical Sciences Nature Patients Performance Pharmaceutical Preparations Pharmacologic Substance Pharmacology Phase Phosphotransferases Play Process Production Protein Conformation Protein Footprinting Proteins Proteome Proteomics Reaction Reproducibility Research Role Safety Signal Transduction Small Business Innovation Research Grant Sodium Solvents Structure Surface Techniques Technology Time Toxin Transmembrane Domain Universities Washington Work adverse drug reaction analytical tool aqueous base detector dosimetry druggable target experimental study improved innovation interest new technology novel pathogen pathogenic virus professor programs research and development tool

项目摘要

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.

Gennext I期Sbir子序列标题为“多波长荧光激进剂量测定法，用于实时评估蛋白质足迹的自由基产量”是响应对高阶新工具的敏锐的需求生物药物和膜蛋白靶标的结构分析（HOS）。新兴的HOS分析技术是羟基自由基（HRPF）。 HRPF 涉及与羟基自由基反应的蛋白质外部的不可逆转标记随后的MS分析以识别蛋白质的外部。我们有开发了执行HRPF的商业解决方案。最近一个新的贵重依赖于oh创建的三氟甲基（TFM）自由基的足迹技术已经开发了三磷酸钠水溶液的根本攻击，这表现出巨大的希望当与HRPF结合使用时。 TFM蛋白足迹（TFMPF）非常完善对于HRPF，由于TFMPF有效地标记了氨基酸保留的，这些氨基酸保留了相对“静音”至哦，激进的攻击。当一起使用时，HRPF和TFMPF提供了大量的覆盖范围并检测溶剂可访问残差，因此代表了变革性改进生物药物HOS评估。 TFMPF的实践是由Michael Gross教授的开创性的华盛顿大学，密苏里州圣路易斯。虽然表现出巨大的承诺，可以解决未满足药物研究中的挑战，TFMPF的可重复性受到挑战背景清除的差异。与总实验室合作，我们的工作将把我们创新的HRPF激进剂量学技术扩展到TFMPF。 Gennext Technologies是唯一用于HRPF HOS分析的公司商业化产品。我们的目标是将HRPF和TFMPF流程的合并使用学术研究实验成有价值的分析工具。一旦简化了转变为强大的技术，我们设想了HRPF和 TFMPF启用：副型和表位MAB生物药物与他们的膜靶；阐明铅结合到正构物或变构膜靶；揭示次级信使信号传导级联 GPCR铅化合物；并检测到角度 /变构抗的影响诸如激酶和生长因子等靶标的肿瘤。