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将检验以下假设，即计算设计的蛋白质“小型插头”和逻辑门控开关 靶向脑脊液（CSF）蛋白生物标志物和蛋白质颗粒可以用作多功能捕获和 检测剂可极大地改善CSF样品的分子表征。这些的可用性 试剂应支持U19的总体目标，即提高我们对CSF生物标志物的理解为直接 与年龄相关的认知下降和阿尔茨海默氏病（AD）病理生理学的度量。 我们已经将工作计划纳入了高度重点的U19项目：下一代翻译 阿尔茨海默氏症和相关痴呆症的蛋白质组学检验上述假设。与项目合作 1–3，我们的主要目标是开发一套优化的计算设计的小型插头（Hyperstable Binding 长度<65 aa）作为CSF蛋白生物标志物捕获剂和超特异性逻辑门控试剂的蛋白质 检测具有两个定义的蛋白质成分的CSF颗粒。 该研究将在计算设计和实验测试之间进行迭代，每个阶段的反馈 从与项目1-3合作进行的CSF测定实验，该实验将指导改进 用于捕获指定CSF生物标志物蛋白靶标的MiniBinder设计方法，以及用于超特异性逻辑 具有两个复合蛋白成分的CSF颗粒的门控检测。结果将是（i）特定的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） 基于新蛋白质的快速需求工程的集成计算实验管道 一般来说，神经退行性疾病的诊断剂。因此，项目4依靠结束 高度交互式U19计划中与项目1-3和核心1-4的合作。