SAMPLE CORE

核心样本

• 批准号: 8694146
• 负责人: Teresa Whei-Mei Fan
• 金额: $ 23.32万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-11 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8694146
• 关键词: A549; Address; Aldehydes; Ammonium; Animals; Aspirate substance; Base Pairing; Biochemical; CD3 Antigens; Cell Extracts; Cell Separation; Cells; Chromatography; Client; Computer software; Consult; Coupled; Crude Extracts; Cultured Cells; Detection; Development; Diagnosis; Disease; Event; Factor V; Fourier Transform; Fourier transform ion cyclotron resonance; Fractionation; Funding; Glycols; Goals; Human Resources; Informatics; Infusion procedures; Instruction; Internet; Ions; Isomerism; Isotopes; Ketones; Label; Leukocytes; Link; Lipids; Lymphocyte; Magnetic Resonance Imaging; Malignant Neoplasms; Mass Spectrum Analysis; Metabolic; Methods; Microfluidics; Molecular; Mus; NMR Spectroscopy; Nipples; Nucleotides; Parents; Pattern; Phase; Plasma; Population; Preparation; Process; Proteins; Protocols documentation; Reaction; Recovery; Reproducibility; Research; Resources; Rodent; Sample Size; Sampling; Scheme; Secure; Services; Solvents; Specimen; Speed; Structure; Subcellular Fractions; Sulfhydryl Compounds; Surface; Technology; Testing; Tissues; Urine; Weight; Work; adduct; base; biological systems; design; functional group; granulocyte; high throughput analysis; human disease; human tissue; improved; in vivo; instrument; macrophage; meetings; metabolomics; nano-electrospray; operation; particle; prevent; sound; stable isotope; tandem mass spectrometry; tool; ultra high resolution

PROJECT SUMMARY (See instructions): Recent advances in metabolomics technologies have made possible simultaneous analysis of hundreds of metabolites by mass spectrometry (MS) and nuclear magnetic resonance spectroscopy (NMR). As the "gateway" to meaningful metabolomics analysis, sample processing has critical issues not widely addressed in metabolomics studies, such as 1) sample integrity; 2) reproducibility and range of metabolite recovery; 3) tools for extending metabolite coverage, enabling high throughput & robust metabolite analysis, and facilitating de novo structure elucidation of unknowns. Our Center (CREAM) has developed test-proven protocols for processing a wide selection of biospecimens with the aim of maintaining biochemical integrity and reproducible recovery of metabolites in large-scale. These protocols have been applied to many stable isotope-resolved metabolomics (SIRM) studies to reveal key reprogrammed metabolic events in human diseases. We have also shown the utility of chemoselective (CS) tagging in enriching low-level, labile, and volatile carbonyl metabolites for high throughput analysis by direct infusion Fourier transform-ion cyclotron resonance-mass spectrometry (FTICR-MS). Our protocols can be further enhanced in terms of throughput and coverage of previously inaccessible metabolites. We will support sample processing activities for the U24 RCMRC with three Specific Aims: SA1. To process a wide range of biospecimens researched by Clients. These range from cells, tissues, media, biofluids to animals. We have provided this service since 2007 and will greatly expand the operation with added resources including sample preparation for in vivo NMR. SA2. To implement microfluidics-based subcellular/cell separation and metabolite extraction. We will test and refine microfiuidic platforms for separation of subcellular particles, cell sorting, and metabolite extraction to offer more efficient, speedy, and high throughput sample processing than existing methods. SA3. To implement profiling and multiplexed quantification of metabolites and their labeling patterns by FTICR- MS in a wide range of biological systems using chemoselective (CS) tagging of functional groups (FG). We will implement the CS capture approach (funded by a separate Common Funds R01) for enriching previously inaccessible metabolites with carbonyl, amino, thiol, and diol FG for robust identification and multiplexed analysis by FTICR-MS and NMR.

项目摘要（请参阅说明）：代谢组学技术的最新进展使通过质谱法（MS）和核磁共振光谱（NMR）同时分析了数百种代谢产物。作为有意义的代谢组学分析的“网关”，样本处理具有在代谢组学研究中未广泛解决的关键问题，例如1）样本完整性； 2）可重复性和代谢物回收范围； 3）用于扩展代谢物覆盖范围的工具，实现高吞吐量和稳健的代谢物分析，并促进从头结构阐明未知数。我们的中心（Cream）开发了用于处理多种生物测量的测试经验方案，目的是维持生化完整性和大规模代谢物的可再现回收率。这些方案已应用于许多稳定的同位素分辨代谢组学（SIRM）研究，以揭示人类疾病中的重编代谢事件。我们还显示了化学选择性（CS）标记在富集低水平，不稳定和挥发性羰基代谢物方面通过直接输注傅立叶傅立叶旋转性环绕resonance-sas-Mass光谱法（FTICR-MS）进行高吞吐量分析的实用性。我们的方案可以进一步增强以前无法访问的代谢物的吞吐量和覆盖范围。我们将支持U24 RCMRC的样本处理活动，具有三个特定目标：SA1。处理客户研究的广泛的生物测量。这些范围从细胞，组织，培养基，生物流体到动物。自2007年以来，我们提供了这项服务，并将通过增加资源（包括Vivo NMR的样本准备）大大扩展运营。 SA2。实施基于微流体的亚细胞/细胞分离和代谢物提取。我们将测试和完善缩微平台，以分离亚细胞颗粒，细胞分选和代谢物提取，以提供比现有方法更有效，快速和高吞吐量样品处理。 SA3。使用官能团（FG）的化学选择性（CS）标记在广泛的生物系统中，通过FTICR-MS实施代谢物及其标记模式的分析和多路复用定量​​。我们将实施CS捕获方法（由单独的共同基金R01资助），以用羰基，氨基，硫醇和DIOL FG富集以前无法访问的代谢物，以通过FTICR-MS和NMR进行强大的识别和多重分析。