Project 4: Novel reagent development to enable molecular characterization

项目 4：开发新型试剂以实现分子表征

• 批准号: 10359195
• 负责人: DAVID BAKER
• 金额: $ 44.88万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-15 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10359195
• 关键词: Affinity; Age-associated memory impairment; Algorithms; Alzheimer' s Disease; Alzheimer' s disease related dementia; Amino Acids; Amyloid; Amyloid beta-Protein; Antibodies; Binding; Binding Proteins; Biological Assay; Biological Markers; Brain-Derived Neurotrophic Factor; Cerebrospinal Fluid; Cerebrospinal Fluid Proteins; Clinical; Collaborations; Complement; Data Storage and Retrieval; Detection; Development; Diagnostic; Engineering; Feedback; Functional disorder; Goals; Green Fluorescent Proteins; Length; Logic; Measures; Methods; Molecular; Monoclonal Antibodies; Neurodegenerative Disorders; Outcome; Parkinson Disease; Peptides; Production; Protein Engineering; Proteins; Reagent; Research; Resources; Sampling; Series; Specific qualifier value; Specificity; Statistical Data Interpretation; Surface; System; Testing; Work; age related neurodegeneration; alpha synuclein; base; data dissemination; data sharing; design; detection platform; experimental study; improved; neurogranin; next generation; novel; particle; programs; protein biomarkers; targeted biomarker; translational proteomics

Abstract Project 4 will test the hypothesis that computationally designed protein “minibinders” and logic-gated switches targeting cerebrospinal fluid (CSF) protein biomarkers and protein particles can serve as versatile capture and detection agents to vastly improve the molecular characterization of CSF samples. The availability of these reagents should support the overall U19 goal of improving our understanding of CSF biomarkers as direct measures of age-related cognitive decline and Alzheimer's Disease (AD) pathophysiology. We have integrated our work plan within the highly focused U19 Project: Next Generation Translational Proteomics for Alzheimer's and Related Dementias to test the above hypothesis. In collaboration with Projects 1–3, our main goal is to develop an optimized set of computationally designed minibinders (hyperstable binding proteins of length <65 aa) as CSF protein biomarker capture agents, and ultra-specific logic-gated reagents for detection of CSF particles that have two defined protein components. The research will iterate between computational design and experimental testing, with feedback at each stage from CSF assay experiments conducted in collaboration with Projects 1-3 which will guide improvement of the minibinder design methods for capture of specified CSF biomarker protein targets, and for ultra-specific logic- gated detection of CSF particles with two composite protein components. The outcomes will be (i) specific CSF biomarker capture and detection systems for AD and other age-related neurodegenerative disorders, and (ii) an integrated computational-experimental pipeline for rapid on demand engineering of new protein based diagnostic agents for neurodegenerative disorders in general. Therefore, Project 4 relies on a close collaboration with Projects 1-3 and Cores 1-4 within the highly interactive U19 program.

抽象的 项目 4 将测试通过计算设计蛋白质“微型结合器”和逻辑门控开关的假设 靶向脑脊液 (CSF) 蛋白质生物标志物和蛋白质颗粒可作为多功能捕获和 检测剂可大大改善脑脊液样本的分子表征。这些的可用性 试剂应支持 U19 的总体目标，即提高我们对 CSF 生物标志物的理解作为直接 与年龄相关的认知能力下降和阿尔茨海默病（AD）病理生理学的测量。 我们已将我们的工作计划纳入高度集中的 U19 项目：下一代转化 阿尔茨海默病和相关痴呆症的蛋白质组学来检验上述假设。与项目合作 1-3，我们的主要目标是开发一组优化的计算设计的迷你绑定器（超稳定绑定 长度 <65 个氨基酸的蛋白质）作为 CSF 蛋白质生物标志物捕获剂，以及超特异性逻辑门控试剂 检测具有两种确定的蛋白质成分的脑脊液颗粒。 该研究将在计算设计和实验测试之间迭代，并在每个阶段提供反馈 来自与项目 1-3 合作进行的 CSF 测定实验，该实验将指导改进 用于捕获特定 CSF 生物标志物蛋白质靶标以及超特异性逻辑的微型结合剂设计方法 具有两种复合蛋白质成分的脑脊液颗粒的门控检测。结果将是 (i) 具体的 CSF AD 和其他与年龄相关的神经退行性疾病的生物标志物捕获和检测系统，以及 (ii) 一个集成的计算实验管道，用于基于新蛋白质的快速按需工程 一般神经退行性疾病的诊断剂。因此，项目 4 依赖于一个封闭的 与高度互动的 U19 计划中的项目 1-3 和核心 1-4 进行合作。