Sample Pretreatment for Human Fluids

人体体液样品预处理

• 批准号: 8715324
• 负责人: Frank Jahnke
• 金额: $ 26.59万
• 依托单位: SONATA BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-08 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8715324
• 关键词: Acids; Acrylamides; Adsorption; Affect; Affinity; Ampholyte Mixtures; Antibodies; Area; Attention; Benchmarking; Biological Assay; Biological Markers; Blood Plasma Volume; Cancer Patient; Cells; Charge; Clinic; Clinical; Companions; Complex; Couples; DNA; Development; Devices; Diagnostic; Diagnostic tests; Disease; Electrolytes; Electrophoresis; Equation; Equilibrium; Eye; Gel; Goals; Health; High Pressure Liquid Chromatography; Human; Industry; Isoelectric Focusing; Isoelectric Point; Left; Liquid Chromatography; Liquid substance; Malignant Neoplasms; Malignant neoplasm of lung; Mass Spectrum Analysis; Measurement; Measures; Membrane; Methods; Mutation; Organic solvent product; Organism; Pathway interactions; Patients; Peptide Library; Peptides; Pharmacologic Substance; Phase; Plasma; Precipitation; Process; Proteins; Proteome; Proteomics; RNA; Reagent; Research; Research Infrastructure; Residual volume; Sample Size; Sampling; Sodium Chloride; Solid; Solutions; Solvents; Stratification; Surface; Testing; Tissues; Two-Dimensional Gel Electrophoresis; Vacuum; Variant; Work; base; clinically relevant; cohort; combinatorial; cost; gel electrophoresis; instrument; internal control; novel; physical property; surfactant; tandem mass spectrometry; tool; trend

DESCRIPTION (provided by applicant): In this proposal we seek to develop a new method and instrument to reduce the complexity of protein mixtures derived from human fluids with an eye to discovery of very low-abundance protein biomarkers. The proposed method accomplishes two tasks: it reduces the dynamic range of the protein mixture by three or more orders of magnitude and it concentrates proteins by a factor of 100 or more so that low- abundance can be detected and concentration differences noted. The method does not use antibodies, combinatorial peptide libraries, carrier ampholytes or solid-liquid adsorption equilibria such as HPLC. The samples produced are compatible with 2D gel electrophoresis and 2D liquid chromatography tandem mass spectrometry approaches. By using different physical methods to separate the protein mixtures, the goal is to uncover new biomarkers that have eluded existing state-of-the-art methods. After development, the proposed method and instrument will be demonstrated using human plasma, where a sample from a lung cancer patient is compared with a reference that does not have cancer. Additional low-abundance proteins that are potential biomarkers will be uncovered when compared with traditional immunodepletion methods.

描述（由申请人提供）：在该提案中，我们寻求开发一种新的方法和仪器，以降低源自人类液体的蛋白质混合物的复杂性，着眼于发现丰度非常低的蛋白质生物标志物。所提出的方法完成了两个任务：它将蛋白质混合物的动态范围降低了三个或更多个数量级，并且它将蛋白质浓缩了100倍或更多倍，使得可以检测到低丰度并注意到浓度差异。该方法不使用抗体、组合肽文库、载体两性电解质或固液吸附平衡如HPLC。所产生的样品与2D凝胶电泳和2D液相色谱串联质谱法兼容。通过使用不同的物理方法来分离蛋白质混合物，目标是发现新的生物标志物，这些生物标志物避开了现有的最先进的方法。开发完成后，将使用人血浆证明所提出的方法和仪器，其中将来自肺癌患者的样本与未患癌症的参考进行比较。与传统的免疫耗竭方法相比，将发现作为潜在生物标志物的其他低丰度蛋白质。