Resource for the Development of Biomedical Accelerator Mass Spectrometry (AMS)

生物医学加速器质谱 (AMS) 开发资源

• 批准号: 8666887
• 负责人: Kenneth W. Turteltaub
• 金额: $ 196.64万
• 依托单位: UNIVERSITY OF CALIF-LAWRNC LVRMR NAT LAB
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8666887
• 关键词: Animals; Area; Automobile Driving; Biological; Biological Markers; Biological Process; Biology; Blood; Body Fluids; Cells; Chemicals; Complex; Development; Dose; Enzymes; Formalin; Funding; Genetic Materials; Human; Label; Life; Ligand Binding; Liquid substance; Measurement; Measures; Mediating; Medical; Medicine; Metabolism; Methods; Modeling; Modification; Nucleic Acids; Organism; Paper; Patients; Pharmaceutical Preparations; Physiologic pulse; Play; Post-Translational Protein Processing; Procedures; Proteins; Reaction; Research; Research Personnel; Resource Development; Resources; Role; Sampling; Source; System; Systems Biology; Techniques; Technology; Tissues; United States National Institutes of Health; Work; Writing; Xenobiotics; accelerator mass spectrometry; adduct; analytical method; anthropogenesis; base; cell type; chemical reaction; design; interest; macromolecule; milliliter; receptor; research study; small molecule; synthetic protein; tissue fixing; tissue processing

DESCRIPTION (provided by applicant): Since its inception in 1999, the NIH-funded Research Resource on the biomedical applications of accelerator mass spectrometry (AMS) has performed numerous groundbreaking studies in many areas including biomolecule turnover, quantitative metabolism of endogenous compounds and xenobiotics, and analysis of potentially mutagenic adducts. These studies were made possible by the high sensitivity, accuracy, and precision of AMS. The work proposed in this TR&D core will build on this legacy of successful, high impact research and will extend the applicability of current AMS experimental and analytical methods to a wider set of scientific and technical challenges. The focus for this core i the development and routine implementation of methods for absolute quantitation of proteins and other biological macromolecules. This core will support several driving biomedical projects, including: 1) Characterization of turnover rates of tissues and cell types labeled as a result of anthropogenic atmospheric 14C release (bomb-pulse biology); 2) Absolute quantitation of protein post-translational modifications; 3) Identification of receptors based on 14C-ligand binding; and 4) Development of labeling strategies, separation techniques, and analytical procedures for quantitation of computationally-designed synthetic protein drugs in biological matrices, in support of animal dosing experiments and human microdosing studies. Biological macromolecules play a wide variety of essential roles in cells, tissues, and organisms. Proteins are the basic building blocks of all living organisms, and proteinaceous enzymes catalyze the chemical reactions responsible for life. Nucleic acids provide the genetic material containing information for building, maintaining, and regulating living organisms. Because biological macromolecules play such essential and diverse roles, improvements in our ability to quantitatively trace macromolecules and their modifications will enable important discoveries that will significantly advance our understanding of biological processes. In a recent paper, Hanke et al (Hanke et al., 2008) wrote: "Ultimately, it would be highly desirable to obtain exact quantitative values of each protein in a system, e.g., their copy number per cell or their concentration in nanogram per milliliter of body fluid. While this kind of basic information about a protein is already per se valuable for the biologist, systems biology even requires it as input for modeling. In a medical context, knowing the exact amounts of certain proteins in blood or other common sources of biomarkers can provide diagnostically relevant information for patient treatment." Analytical measurement of proteins and other biological macromolecules presents a number of significant technical challenges, particularly when quantitation is required. Macromolecule extraction from biological matrices is typically more complex than the extraction of small molecules, and extracts are more prone to contamination and degradation. Macromolecules such as proteins and nucleic acids are heterogeneous molecules, and may be chemically modified as a result of enzyme-mediated reactions (e.g., post-translational modifications) or as a result of tissue processing, as in the case of formalin-fixed tissue. In addition, biological macromolecules may be susceptible to cleavage by physical or chemical means. While analytical standards can usually be purchased or synthesized for small molecules of interest, similar standards are rarely available for biological macromolecules. The ability to measure absolute quantities of biological macromolecules has broad relevance for biology and medicine, as there currently is no standardized, universal analytical method capable of making these measurements. The major focus of this core is to develop methods for applying AMS technology, especially in conjunction with the new liquid sample AMS interface, to overcome these major challenges of biological macromolecule analysis. To this end, we propose to interact with collaborating researchers to solve a variety of important biological problems.

描述（由申请人提供）：自 1999 年成立以来，NIH 资助的加速器质谱 (AMS) 生物医学应用研究资源已在许多领域进行了大量开创性研究，包括生物分子周转、内源化合物和外源物质的定量代谢以及潜在诱变加合物的分析。 AMS 的高灵敏度、准确性和精密度使这些研究成为可能。该 TR&D 核心中提出的工作将建立在这一成功的高影响力研究的基础上，并将当前 AMS 实验和分析方法的适用性扩展到更广泛的科学和技术挑战。该核心的重点是蛋白质和其他生物大分子绝对定量方法的开发和常规实施。该核心将支持多个驱动性生物医学项目，包括： 1) 表征由于人为大气 14C 释放而标记的组织和细胞类型的周转率（炸弹脉冲生物学）； 2）蛋白质翻译后修饰的绝对定量； 3) 基于14C-配体结合的受体鉴定； 4) 开发用于定量生物基质中计算设计的合成蛋白药物的标记策略、分离技术和分析程序，以支持动物给药实验和人体微剂量研究。 生物大分子在细胞、组织和生物体中发挥着多种重要作用。蛋白质是所有生物体的基本组成部分，蛋白质酶催化生命的化学反应。核酸提供含有构建、维持和调节生物体信息的遗传物质。由于生物大分子发挥着如此重要和多样化的作用，因此我们定量追踪大分子及其修饰的能力的提高将带来重要的发现，从而显着增进我们对生物过程的理解。在一个 汉克等人最近的论文（Hanke et al., 2008）写道： “最终，非常希望获得系统中每种蛋白质的精确定量值，例如每个细胞的拷贝数或每毫升体液的纳克浓度。虽然这种蛋白质的基本信息本身对于生物学家来说已经很有价值，但系统生物学甚至需要它作为建模的输入。在医学背景下，了解血液或其他常见生物标志物来源中某些蛋白质的确切含量可以提供 患者治疗的诊断相关信息。” 蛋白质和其他生物大分子的分析测量提出了许多重大的技术挑战，特别是在需要定量时。从生物基质中提取大分子通常比小分子提取更复杂，并且提取物更容易受到污染和降解。诸如蛋白质和核酸之类的大分子是异质分子，并且可能由于酶介导的反应（例如翻译后反应）而被化学修饰。 修改）或作为组织处理的结果，如福尔马林固定组织的情况。此外，生物大分子可能容易被物理或化学手段裂解。虽然通常可以购买或合成感兴趣的小分子的分析标准品，但很少可用于生物大分子的类似标准品。测量生物大分子绝对量的能力与生物学和医学具有广泛的相关性，因为目前没有能够进行这些测量的标准化、通用的分析方法。该核心的主要重点是开发应用 AMS 的方法 技术，特别是与新的液体样品 AMS 接口相结合，可以克服生物大分子分析的这些主要挑战。为此，我们建议与合作研究人员进行互动，以解决各种重要的生物学问题。