Integrated Top-Down/Bottom-Up Comprehensive Proteomics

集成的自上而下/自下而上的综合蛋白质组学

基本信息

批准号：
7278310
负责人：
Satya Prakash Saxena
金额：
$ 84.35万
依托单位：
CALIBRANT BIOSYSTEMS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-04-07 至 2009-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7278310
关键词：
Address Amino Acid Sequence Biological Blood capillaries Capillary Electrophoresis Caseins Cells Chromatography Codon Nucleotides Complex Condition Coupled Coupling Detection Development Digestion Disease Early Diagnosis Electrospray Ionization Equipment Evaluation Figs - dietary Fluorescence Foundations Fractionation Fungal Genome Gel Genome Genomics Grouping Head Hour Injection of therapeutic agent Isoelectric Point Liquid Chromatography Mass Spectrum Analysis Measurement Measures Membrane Membrane Proteins Methodology Methods Modeling Modification Molecular Monitor Numbers Open Reading Frames Operative Surgical Procedures Organism Pancreatic ribonuclease Peptide Fragments Peptide Sequence Determination Peptides Performance Phase Phase I Clinical Trials Polyacrylamide Gel Electrophoresis Population Post-Translational Protein Processing Preclinical Drug Evaluation Procedures Process Protein Analysis Protein Dynamics Protein Sequence Analysis Proteins Proteolytic Processing Proteome Proteomics Pump Range Regulation Reporting Reproducibility of Results Research Resolution Running Sampling Shotguns Sodium Dodecyl Sulfate Solid Solutions Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Syringes System Systems Biology Techniques Technology Time Tissue Sample Tissues Translating Translations Trypsin Two-Dimensional Polyacrylamide Gel Electrophoresis Yeasts base capillary electric field gel electrophoresis gene function genetic regulatory protein improved indexing information gathering mRNA Differential Displays mass spectrometer molecular mass nanoscale numb protein protein expression protein structure size tandem mass spectrometry tool yeast protein

项目摘要

DESCRIPTION (provided by applicant): Phase I: The R43 phase of this project aims to develop and demonstrate a capillary gel electrophoresis (CGE) based multidimensional protein separation platform capable of providing significant analyte concentration and extremely high resolving power for handling complex protein mixtures prior to mass spectrometry detection. In comparison with conventional SDS-PAGE-based proteome approaches, the proposed CGE/capillary reversed-phase liquid chromatography (CRPLC) proteome technology will be highly automated and offer robust and high throughput fractionation of whole proteins for top-down proteomics while avoiding analyte dilution and loss in an integrated platform. By coupling with a nanoscale trypsin membrane reactor, the ultrafast proteolytic digestion of proteins resolved and eluted from the CGE-based separations enables the combined top-down/bottom-up characterization of post-translational modifications in complex proteomes. Research efforts in the R43 phase will address early implementation, optimization, and demonstration of the proposed proteome technology using yeast cell lysates.

描述（由申请人提供）：第一阶段：该项目的 R43 阶段旨在开发和演示基于毛细管凝胶电泳 (CGE) 的多维蛋白质分离平台，该平台能够提供显着的分析物浓度和极高的分辨率，用于事先处理复杂的蛋白质混合物。至质谱检测。与传统的基于 SDS-PAGE 的蛋白质组方法相比，所提出的 CGE/毛细管反相液相色谱 (CRPLC) 蛋白质组技术将高度自动化，并为自上而下的蛋白质组学提供完整蛋白质的稳健和高通量分级分离，同时避免分析物稀释以及集成平台的损失。通过与纳米级胰蛋白酶膜反应器耦合，对从基于 CGE 的分离中解析和洗脱的蛋白质进行超快蛋白水解消化，能够对复杂蛋白质组中的翻译后修饰进行自上而下/自下而上的组合表征。 R43 阶段的研究工作将解决所提出的使用酵母细胞裂解物的蛋白质组技术的早期实施、优化和演示。