Project 4: Novel reagent development to enable molecular characterization

摘要 项目4将测试假设，计算设计的蛋白质“minibinders”和逻辑门控开关 靶向脑脊液（CSF）蛋白质生物标志物和蛋白质颗粒可用作通用捕获和 检测试剂，以极大地改善CSF样品的分子表征。网站或资源的可用性 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， 与年龄相关的认知衰退和阿尔茨海默病（AD）病理生理学的测量。 我们已经将我们的工作计划整合到高度集中的U19项目中：下一代翻译 阿尔茨海默氏症和相关痴呆症的蛋白质组学来验证上述假设。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minibinders（超稳定绑定 长度<65 aa的蛋白质）作为CSF蛋白质生物标志物捕获剂，以及用于 检测具有两种确定的蛋白质组分的CSF颗粒。 研究将在计算设计和实验测试之间进行，每个阶段都有反馈 从与项目1-3合作进行的CSF测定实验中， 用于捕获特定CSF生物标志物蛋白靶点和用于超特异性逻辑分析的微结合物设计方法 具有两种复合蛋白组分的CSF颗粒的门控检测。结局将是（i）特定CSF 用于AD和其他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，和（ii） 一个集成的计算实验管道，用于快速按需工程化新蛋白质， 一般用于神经变性疾病的诊断剂。因此，项目4依赖于关闭 在高度互动的U19项目中与项目1-3和核心1-4合作。

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.