Project 4: Novel reagent development to enable molecular characterization

摘要 项目4将测试这样一种假设，即通过计算设计的蛋白质“迷你粘合剂”和逻辑门控开关 靶向脑脊液(CSF)蛋白质生物标记物和蛋白质颗粒可用作多功能捕获和 检测试剂极大地改善了脑脊液样本的分子特征。这些产品的可用性 试剂应该支持U19的总体目标，即提高我们对脑脊液生物标志物的直接理解 测量与年龄相关的认知衰退和阿尔茨海默病(AD)的病理生理学。 我们已经将我们的工作计划整合到高度关注的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。与项目协作 1-3，我们的主要目标是开发一套通过计算设计的优化迷你粘合剂(超稳定结合 作为脑脊液蛋白质生物标记物捕捉剂的长度为65aa的蛋白质，以及用于 检测具有两种已定义蛋白质组分的脑脊液颗粒。 这项研究将在计算设计和实验测试之间反复进行，每个阶段都有反馈 来自与项目1-3合作进行的脑脊液分析实验，该实验将指导改进 用于捕获特定脑脊液生物标记物蛋白靶标的微型粘合剂设计方法，以及用于超特定逻辑的 具有两种复合蛋白成分的脑脊液颗粒的门控检测。结果将是(I)特定的脑脊液 AD和其他与年龄相关的神经退行性疾病的生物标记物捕获和检测系统，以及(Ii) 一种用于新型蛋白质按需快速工程的计算-实验一体化流水线 一般神经退行性疾病的诊断剂。因此，项目4依赖于结束 与高度互动的U19计划中的项目1-3和核心1-4进行协作。

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.