Novel Micro-X-ray Translucent Flow Cell Technology for Proteomics, a Broadly Appl

适用于蛋白质组学的新型微 X 射线半透明流动池技术，应用广泛

• 批准号: 7929416
• 负责人: Benjamin P Warner
• 金额: $ 17.89万
• 依托单位: ICAGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7929416
• 关键词: Academia; Address; Albumins; Attention; Binding; Biological Markers; Businesses; Calibration; Cells; Collaborations; Data; Detection; Disease; Drug Interactions; Drug Labeling; Elements; Fluorescence; Funding; Genes; Glycine decarboxylase; Goals; Healthcare; Human Genome; Human Genome Project; Incidence; Kinetics; Label; Laboratories; Lead; Legal patent; Measurable; Measurement; Measures; Methods; Molecular; Pharmaceutical Preparations; Pharmacogenetics; Pharmacologic Substance; Phase; Post-Translational Protein Processing; Preparation; Procedures; Property; Protein Binding; Protein Chemistry; Protein Kinase; Proteins; Proteomics; Public Health; Qualifying; Quantitative Chemistry; Reaction; Reagent; Reporting; Research; Research Personnel; Roentgen Rays; Sampling; Scientist; Series; Solutions; Surface; Surface Plasmon Resonance; System; Techniques; Technology; Testing; Time; Transcript; United States National Institutes of Health; Work; abstracting; design; inhibitor/antagonist; innovation; instrument; new technology; novel; phrases; protein protein interaction; small molecule; sound; technology development; tool

Project Summary/Abstract Caldera Pharmaceutical¿s aim is to develop a broadly applicable, quantitative research tool which addresses core technical challenges in the field of proteomics, including protein-drug interactions, protein-protein interactions, and post-translational modifications. We propose to create a new proteomics technology involving a novel X- ray translucent flow cell design. If our aims are achieved, more accurate, sensitive, and affordable, high-throughput proteomics methods will be available. Our goals are: 1. recognition of protein interactions, and 2. characterization of post- translational modifications. Our long-term objective is to solve key and fundamental problems with proteomic technologies, as described by the world¿s leading scientists. There is a desperate public health need for proteomics, quantitative chemistry, pharmacogenetics, and others to contribute to reducing the incidence of adverse treatment reactions, discovering diseases earlier, and optimizing and individualizing treatments. We intend to focus on the development of technologies that will permit observations of unaltered small molecules which are quantitative and made in real time. Real time measurements can be made using Caldera¿s technology because reactions are performed in solution. Caldera Pharmaceuticals, in collaboration with Los Alamos National Laboratory (LANL), developed Reagentless Pharmacoproteomic Measurement (RPM), a patented technology which uses micro-X-ray fluorescence to quantify the binding of drugs to proteins. It is a unique label-free system. Currently, the binding kinetics of only a few proteins and their inhibitors can be measured accurately. Most proteins do not have a readily measurable reaction product (as is the case for protein kinases), so these proteins remain very difficult to test. Thus, even though tethering a protein [as with surface plasmon resonance (SPR) systems] can lead to incorrect binding constants by orders-of-magnitude, most pharmaceutical companies have purchased SPR instruments. With Caldera¿s proposed innovation, researchers will have a major technological advantage, for the first time; truly label-free binding measurements available for all proteins and most drugs. Furthermore, our technology has the potential to provide benefit to all biomedical and pharmaceutical research which involves drug to protein binding. The proposed project is a method and apparatus for rapidly characterizing proteins for their binding properties and post-translational modifications. This characterization will be achieved using X-ray fluorescence measurements of proteins, their post-translational modifications, and any binding partners (drugs, proteins, co-factors). These measurements use native, unmodified proteins and reagents without the need either to label the drug or to tether the protein. This measurement technique eliminates the experimental biases introduced by existing technologies. The primary experimental biases to be removed are labeling and albumin elimination. The significance of Caldera¿s innovative tool is that determining protein characterization currently requires significant sample preparations. For example, measuring drug-protein binding kinetics has previously required labeling of the drug and/or tethering of the protein to a surface. Both of these procedures render kinetic data unreliable or incorrect (slower rates and other constants off by as much as four orders of magnitude).

项目概要/摘要 卡尔德拉制药公司¿的目标是开发一种广泛适用的定量研究 解决蛋白质组学领域核心技术挑战的工具，包括 蛋白质-药物相互作用、蛋白质-蛋白质相互作用和翻译后 我们建议创建一种新的蛋白质组学技术，涉及一种新颖的 X-。 如果我们的目的达到了，射线半透明的流通池设计更加准确、灵敏、 并且将提供负担得起的高通量蛋白质组学方法。 我们的目标是：1. 识别蛋白质相互作用，2. 表征后蛋白相互作用 我们的长期目标是解决关键和根本问题。 正如世界所描述的那样，蛋白质组技术存在的问题¿领先的科学家。 公共卫生迫切需要蛋白质组学、定量化学、 药物遗传学等有助于减少不良反应的发生 治疗反应，及早发现疾病，优化和个体化 我们打算专注于开发能够实现治疗的技术。 对未改变的小分子的观察是定量的并且是真实的 可以使用 Caldera 进行实时测量。的技术因为 Caldera Pharmaceuticals 与 Los 合作在溶液中进行反应。 阿拉莫斯国家实验室 (LANL)，开发出无试剂药物蛋白质组学 测量 (RPM)，一项专利技术，利用微 X 射线荧光来测量 量化药物与蛋白质的结合是目前独特的无标记系统。 只有少数蛋白质及其抑制剂的结合动力学可以准确测量。 大多数蛋白质没有易于测量的反应产物（例如 蛋白激酶），因此这些蛋白质仍然很难测试。 束缚蛋白质[如表面等离子共振（SPR）系统]可能会导致 大多数制药公司的结合常数存在数量级错误 已经购买了 Caldera 的 SPR 仪器。提出的创新，研究人员将 拥有重大技术优势，首次实现真正的无标签装订； 此外，我们的测量可用于所有蛋白质和大多数药物。 技术有潜力为所有生物医学和制药领域带来益处 涉及药物与蛋白质结合的研究。 拟议的项目是一种快速表征蛋白质的方法和装置 它们的结合特性和翻译后修饰将是。 使用蛋白质的 X 射线荧光测量实现，其翻译后 修饰以及任何结合伴侣（药物、蛋白质、辅因子）。 测量使用天然的、未修饰的蛋白质和试剂，无需 这种测量技术消除了药物标记或束缚蛋白质。 现有技术引入的实验偏差。 需要消除的偏差是标记和白蛋白消除。 火山口的重要性??其创新工具是确定蛋白质特征 目前需要大量的样品制备，例如测量药物-蛋白质。 结合动力学以前需要对药物进行标记和/或对药物进行束缚 这两个过程都会导致动力学数据不可靠或不正确。 （较慢的速率和其他常数相差四个数量级）。