National Center for Quantitative Biology of Complex Systems

国家复杂系统定量生物学中心

• 批准号: 10688030
• 负责人: JOSHUA J COON
• 金额: $ 32.79万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-05 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10688030
• 关键词: Address; Biological; Biological Assay; Biology; Biomedical Technology; Cells; Chromatography; Complex; Complex Mixtures; Consumption; Coupling; Data Analyses; Disease; Exclusion; Expeditions; Foundations; Fracture; Gases; Genes; Goals; Heterogeneity; Hour; Human; Hybrids; Instruction; Intelligence; Isomerism; Laboratories; Lipids; Mass Spectrum Analysis; Measurement; Measures; Medicine; Methodology; Methods; Mission; Modernization; Molecular; Monitor; National Center for Research Resources; Noise; Nucleic Acids; Organism; Peptides; Performance; Persons; Phase; Preparation; Protein Analysis; Proteins; Proteome; Proteomics; Reproducibility; Resolution; Resources; Running; Sampling; Signal Transduction; Source; Structure; System; Techniques; Technology; Time; Transcript; Translating; Work; Yeasts; adduct; computerized data processing; data acquisition; dimer; equipment acquisition; high throughput technology; human genome sequencing; improved; innovation; innovative technologies; instrument; ion mobility; lipidome; lipidomics; liquid chromatography mass spectrometry; mass spectrometer; metabolome; metabolomics; multiple omics; nano-liquid chromatography; prevent; success; tandem mass spectrometry; tool; ultra high pressure

PROJECT SUMMARY – TR&D 3 MS is a popular and powerful technique for measuring proteomes, metabolomes, and lipidomes. For each of these classes, the MS analysis is remarkably similar, consisting of chromatographic separation, mass measurement, and tandem MS. Despite this commonality, nearly all MS laboratories specialize in analysis of a single ome, at the exclusion of the others. The integrated workflow we describe will overcome this limitation by providing a chromatography and MS technology capable of separating and characterizing a complex mixture containing peptides, metabolites, and lipids. We envision this to happen in a few hours of analysis time and to provide comprehensive coverage across omes. Here we lay the foundation for this work by eliminating existing shortcomings in proteomics and lipidomic technologies and commence work on an integrated multi-omic platform that allows for simultaneous analysis of peptides, lipids, and metabolites. First, work by us and others has demonstrated that with the latest high MS/MS sampling rates, the primary limitation preventing deeper single- shot proteome analysis is peptide separations. Second, we have identified two primary obstacles limiting lipidomic analyses: (1) Poor separation of isomeric and isobaric lipid species, which often co-elute such that the number of unique lipid species and their quantities cannot be determined. (2) MS/MS bandwidth is limited and often consumed by redundant sampling of in-source fragments, adducts, and dimers. Finally, presently, most multi-omic studies split biological samples so that each omic analysis is conducted separately – from sample preparation to data analysis – by different people with different MS systems. This fragmentation results in substantial hands-on sample preparation labor and instrument acquisition time. Sample-to-sample heterogeneity, independent sample handling, and instrument-to-instrument variance all contribute noise, thereby obscuring otherwise strong biomolecular associations.

项目总结--研发3 MS是测量蛋白质组、代谢体和脂体的一种流行且功能强大的技术。对于每一个 在这些类别中，MS的分析非常相似，包括色谱分离、质量 尽管有这种共性，几乎所有的MS实验室都专门分析一种 唯一的一部分，排除了其他部分。我们描述的集成工作流将通过以下方式克服这一限制 提供一种能够分离和表征复杂混合物的层析和MS技术 含有多肽、代谢物和类脂的。我们设想这将在几个小时的分析时间内发生，并 提供跨OMES的全面覆盖。在这里，我们通过消除现有的 蛋白质组学和脂组学技术方面的缺陷，并开始在综合多组学平台上工作 这允许同时分析多肽、脂类和代谢物。首先，我们和其他人的工作已经 表明，在最新的高MS/MS采样率下，防止更深层次的单次- 蛋白质组分析是多肽的分离。第二，我们确定了两个主要障碍，限制了 脂肪组学分析：(1)同分异构体和等压脂类分离不佳，它们经常共洗脱，从而使 独特的类脂种类的数量和数量无法确定。(2)MS/MS带宽有限， 经常被源内片段、加合物和二聚体的冗余采样所消耗。最后，目前，最 多组研究将生物样本分开，这样每个组分析都是分开进行的--从样本 为数据分析做准备-由不同的人使用不同的MS系统。这种碎片化导致 大量的动手样品准备工作和仪器获取时间。样品到样品 异质性、独立的样品处理和仪器之间的差异都会造成噪声，因此 掩盖了其他强烈的生物分子联系。