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).

项目摘要/摘要 Caldera Pharmaceutical的目的是开发一项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 修改。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- 射线半透明流动池设计。如果我们的目标得以实现，更准确，敏感， 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，以及2。 翻译修改。我们的长期目标是解决关键和基本 蛋白质组学技术的问题，如世界领先的科学家所描述的那样。 迫切需要蛋白质组学，定量化学的需求， 药物遗传学和其他人有助于减少逆境事件 治疗反应，较早发现疾病，优化和个性化 治疗。我们打算专注于将允许的技术的发展 观察不变的小分子，这些分子是定量的，并以真实的形式进行 时间。可以使用Caldera的技术进行实时测量，因为 反应是在溶液中进行的。 Caldera Pharmaceuticals与Los合作 阿拉莫斯国家实验室（LANL）开发了无试剂的药物蛋白质组学 测量（RPM），一种使用Micro-X射线荧光的专利技术 量化药物与蛋白质的结合。这是一个独特的无标签系统。目前， 只能准确测量几种蛋白质及其抑制剂的结合动力学。 大多数蛋白质都不容易测量的反应产物（就像 蛋白激酶），因此这些蛋白质仍然很难测试。即使 将蛋白质束缚（与表面等离子体共振（SPR）系统一样）可能导致 大多数制药公司通过魔术命令的绑定常数不正确 已经购买了SPR仪器。随着Caldera的拟议创新，研究人员将 首次具有主要的技术优势；真正的无标签绑定 用于所有蛋白质和大多数药物的测量。此外，我们的 技术有可能为所有生物医学和药品提供好处 涉及药物与蛋白质结合的研究。 提出的项目是一种快速表征蛋白质的方法和设备 它们的结合特性和翻译后修饰。这个特征将是 使用蛋白质的X射线荧光测量来实现其翻译后 修改和任何结合伴侣（药物，蛋白质，副因素）。这些 测量使用本机，未修饰的蛋白质和试剂，无需 标记药物或束缚蛋白质。这种测量技术消除了 现有技术引入的实验偏见。主要实验 要删除的偏见是标签和相册消除。 Calderaâs创新工具的意义在于确定蛋白质表征 目前需要大量的样本准备。例如，测量药物蛋白 结合动力学先前需要标记药物和/或束缚 蛋白质到表面。这两个过程都使动力学数据不可靠或不正确 （速度较慢，其他常数降低了多达四个数量级）。