Agilent Complete 2D-LC-QTOF System

安捷伦完整的 2D-LC-QTOF 系统

基本信息

批准号：
10797808
负责人：
Ying Ge
金额：
$ 19.99万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-25 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10797808
关键词：
Address Antibodies Biological Assay Biological Markers Blood Cardiac Cardiovascular Diseases Cells Chemistry Code Collaborations Complex Computer software Diabetes Mellitus Diagnosis Disease Disease Progression Drug Targeting Electronics Equipment Extracellular Matrix Face Foundations Funding G Protein-Coupled Receptor Signaling G-Protein-Coupled Receptors Genetic Variation Genomics Heart Diseases Magnetic nanoparticles Mainstreaming Malignant Neoplasms Mass Spectrum Analysis Membrane Methods Molecular Nanotechnology Organic Chemistry Play Post-Translational Protein Processing Productivity Proteins Proteome Proteomics Publications Request for Proposals Research Role Solubility Structure Structure-Activity Relationship Surface System Technology Tissues Troponin I Variant biological systems design diagnostic assay empowerment human disease individual variation innovation insight interdisciplinary approach new therapeutic target novel novel strategies precision medicine success surfactant tool

项目摘要

Summary In the post-genomics era, a comprehensive analysis of “proteoforms” that arise from genetic variations and post-translational modifications (PTMs) is essential for understanding biological systems at a functional level and for dissecting complex molecular systems with consideration of individual variability for precision medicine. Top- down mass spectrometry (MS)-based proteomics that analyzes intact proteins is the most powerful method to comprehensively characterize proteoforms to decipher the PTM codes together with sequence variations. Although significant strides have been made recently in both MS hardware and software to advance top-down MS closer to the mainstream, top-down proteomics still faces major challenges. In particular, the proteome is extremely complex and has a high dynamic range in addition to the low solubility of many proteins, making it highly challenging for high-throughput proteomic study. Building on the success in the last funding period, in this multiple-PI renewal application, we will continue to develop innovative strategies empowered by nanotechnology and materials/organic chemistry to further address the challenges in top-down proteomics. The specific objectives of this proposal are: 1) To address the protein solubility challenge, we will develop a novel strategy enabled by a photocleavable surfactant for extracellular matrix (ECM) proteomics; and design, synthesize, and evaluate a novel class of photocleavable nonionic surfactants that can retain the native structures of proteins for native MS-based top-down proteomics. 2) To address the high dynamic range challenge, we will develop novel surface functionalized magnetic nanoparticles (NPs) to mimic antibodies for capturing and enriching low abundance proteins, such as cardiac troponin I (cTnI, a gold-standard biomarker for heart diseases) from tissues/blood and G-protein coupled receptors (GPCRs, a major class of drug targets) from cells/tissues, for downstream comprehensive analysis of all proteoforms by top-down proteomics. Our highly interdisciplinary approach integrates materials chemistry/nanotechnology with top-down MS-based proteomics, and is based on an existing productive collaboration between two PIs that has led to significant progress and publications from the past funding period. Success in our proposed research will provide innovative tools to enable top-down proteomics of poorly soluble and low abundance proteins, which will lay important technological foundation for understanding the critical role that ECM plays in disease progression in cancer and cardiac diseases, defining the structure-function relationship of native membrane complexes, developing a comprehensive cTnI assay for the diagnosis of cardiac diseases with high accuracy, and understanding the important roles of GPCR signaling during the onset of numerous human diseases including cancer, diabetes, and cardiovascular diseases.

概括后基因组时代，对遗传变异产生的“蛋白质形态”进行全面分析翻译后修饰（PTM）对于在功能水平上理解生物系统至关重要用于解剖复杂的分子系统，考虑精准医学的个体差异。基于质谱 (MS) 的蛋白质组学表明，完整蛋白质是最有效的方法全面表征蛋白质组，以破译 PTM 代码和序列变异。尽管最近在 MS 硬件和软件方面都取得了重大进展，以推进自上而下的发展 MS更接近主流，自上而下的蛋白质组学仍然面临重大挑战，尤其是蛋白质组学。除了许多蛋白质的低溶解度之外，它极其复杂并且具有高动态范围，使其基于上一资助期的成功，本次研究对高通量蛋白质组学研究极具挑战性。多PI更新应用，我们将持续开发纳米技术赋能的创新策略和材料/有机化学，以进一步应对自上而下蛋白质组学的挑战。该提案的目标是：1）为了解决蛋白质溶解度挑战，我们将制定一种新策略由可光裂解的表面活性剂实现细胞外基质 (ECM) 蛋白质组学和设计、合成和评估一类新型可光裂解非离子表面活性剂，该表面活性剂可以保留蛋白质的天然结构基于原生 MS 的自顶向下蛋白质组学 2) 为了解决高动态范围的挑战，我们将开发新颖的蛋白质组学。表面功能化磁性纳米颗粒（NP）可模拟抗体，用于捕获和富集低浓度蛋白质丰度，例如心肌肌钙蛋白 I（cTnI，心脏病的金标准生物标志物）组织/血液和来自细胞/组织的 G 蛋白偶联受体（GPCR，一类主要的药物靶点），通过自上而下的蛋白质组学对所有蛋白质形式进行下游综合分析。方法将材料化学/纳米技术与自上而下的基于 MS 的蛋白质组学相结合，并且基于两位 PI 之间现有的富有成效的合作带来了重大进展并发表了论文我们拟议的研究的成功将提供创新工具，以实现自上而下的目标。难溶性、低丰度蛋白质的蛋白质组学研究，将为研究奠定重要的技术基础了解 ECM 在癌症和心脏病的疾病进展中发挥的关键作用，定义天然膜复合物的结构-功能关系，开发了一种全面的 cTnI 检测方法高精度诊断心脏病，并了解 GPCR 信号传导的重要作用包括癌症、糖尿病和心血管疾病在内的许多人类疾病的发作期间。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Comprehensive characterization of monoclonal antibody by Fourier transform ion cyclotron resonance mass spectrometry.

DOI：
10.1080/19420862.2018.1525253
发表时间：
2019-01
期刊：
mAbs
影响因子：
5.3
作者：
Jin Y;Lin Z;Xu Q;Fu C;Zhang Z;Zhang Q;Pritts WA;Ge Y
通讯作者：
Ge Y

PP2A-B' holoenzyme substrate recognition, regulation and role in cytokinesis.

DOI：
10.1038/celldisc.2017.27
发表时间：
2017
期刊：
Cell discovery
影响因子：
33.5
作者：
Wu CG;Chen H;Guo F;Yadav VK;Mcilwain SJ;Rowse M;Choudhary A;Lin Z;Li Y;Gu T;Zheng A;Xu Q;Lee W;Resch E;Johnson B;Day J;Ge Y;Ong IM;Burkard ME;Ivarsson Y;Xing Y
通讯作者：
Xing Y

A Family of Photolabile Nitroveratryl-Based Surfactants That Self-Assemble into Photodegradable Supramolecular Structures.