Sample Pretreatment for Human Fluids

人体体液样品预处理

基本信息

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
项目类别：
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

项目摘要

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）：在本提案中，我们寻求开发一种新的方法和仪器，以降低源自人体体液的蛋白质混合物的复杂性，着眼于发现非常低丰度的蛋白质生物标志物。所提出的方法完成两项任务：将蛋白质混合物的动态范围降低三个或更多数量级，并将蛋白质浓缩100倍或更多，以便检测低丰度并记录浓度差异。该方法不使用抗体、组合肽库、载体两性电解质或固液吸附平衡（例如 HPLC）。产生的样品与 2D 凝胶电泳和 2D 液相色谱串联质谱方法兼容。通过使用不同的物理方法分离蛋白质混合物，目标是发现现有最先进方法无法发现的新生物标志物。开发完成后，所提出的方法和仪器将使用人类血浆进行演示，其中将来自肺癌患者的样本与未患癌症的参考样本进行比较。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。