High-sensitivity Immunomagnetic System for "Liquid Biopsy" of Alzheimer's Disease

用于阿尔茨海默病“液体活检”的高灵敏度免疫磁系统

• 批准号: 10539340
• 负责人: Hui Mao
• 金额: $ 56.65万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10539340
• 关键词: Address; Adsorption; Alzheimer disease detection; Alzheimer' s Disease; Alzheimer' s disease patient; Alzheimer’s disease biomarker; Amyloid beta-42; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Antibodies; Archives; Attenuated; Biological Markers; Blocking Antibodies; Blood; Blood specimen; Brain; Brain imaging; Cerebrospinal Fluid; Clinical; Clinical Management; Communities; Coupled; Data; Detection; Development; Devices; Diagnosis; Disease Progression; Early Diagnosis; Economic Burden; Enhancement Technology; Enzyme-Linked Immunosorbent Assay; Equipment; Goals; Healthcare; Image; Imaging Techniques; Immunomagnetic Separation; In Vitro; Individual; Ligands; Liquid substance; Magnetic Resonance Imaging; Magnetism; Measurement; Measures; Microfluidic Microchips; Microfluidics; Microspheres; Microtubules; Monitor; Nanotechnology; Noise; Patient Care; Patients; Plasma; Polymers; Population; Population Heterogeneity; Positron-Emission Tomography; Prognostic Marker; Protein Analysis; Proteins; Proteomics; Radioactive; Research; Resources; Risk; Sampling; Scanning; Sensitivity and Specificity; Serum; Specificity; Surface; System; Technology; Testing; Time; abeta deposition; antibody conjugate; biomarker discovery; cost; cost effective; detection platform; diagnostic accuracy; diagnostic biomarker; diagnostic tool; early screening; effective therapy; improved; in-vitro diagnostics; innovation; iron oxide; liquid biopsy; multiplex detection; nano; nanoparticle; nanorod; nanosized; nanotechnology platform; neuroimaging; peripheral blood; point of care; programs; radiotracer; social; surface coating; targeted biomarker; tau Proteins; tau-1

Project Summary Alzheimer’s disease (AD) is a major healthcare and social-economic burden. With effective treatment yet to be developed, the early detection at the prodromal stage and surveillance of the progression are the best approach to control and intervene AD development. While non-invasive positron emission tomography (PET) can measure amyloid-β peptides (Aβs) and microtubule tau proteins in the brain to confirm the onset of AD, the availability of imaging equipment, diagnostic accuracy of single Aβ or tau specific radiotracer and the cost and exposure of radioactive materials limit imaging applications from early screening of large and diverse populations and repeated scans in monitoring disease progression. Therefore, there is an unmet need in developing robust, accessible and cost-effective in vitro diagnostic tools for detecting and measuring the levels of Aβs and tau proteins in cerebrospinal fluid or even blood samples of patients or at-risk individuals at the “point-of-care”. However, serum detection of these AD biomarker demands high sensitivity because of low concentrations of Aβs or tau proteins in blood samples and slow and gradual change of the levels of these biomarkers during disease progression. One major challenge to the current biospecimen based in vitro diagnostics is the biofouling effect, i.e., adsorption of proteins and biomolecules on the surface of the detecting agents and devices. This leads to the formation of the layer of unwanted molecules, which reduces the detection specificity and sensitivity by (1) blocking antibodies to the targeted biomarkers, and (2) causing high “background noise” from adsorbed non-specific proteins and other molecules interfering the detection of targeted biomarkers. We believe that these problems can be overcome by our innovative solutions, i.e., 1) the anti-biofouling polymer to coat nanoparticle capturing agents to block the non-specific protein adsorption, thus protecting the sensitivity and specificity of the targeting ligands/antibodies; 2) high magnetism magnetic iron oxide nanorods (IONRs) with nano-sized magnetic stir bar action to enhance mixing of capturing agents and targeted biomarkers in the microfluidic detection system. By integrating these technologies, our project aims to develop a multiplexing based “liquid biopsy” system with ultra-high sensitivity and specificity for serum detection of selected AD biomarkers, Aβ40, Aβ42, total-tau (tTau), phosphorylated-Tau (pTau)181 and amyloid precursor protein (APP) in one sample. We will make and optimize antibody-conjugated anti-biofouling IONRs with different aspect ratios for highly efficient fluid mixing and multiplex detection of spiked Aβ40, Aβ42, tTau and pTau proteins and APP669-711 in blood samples (Aim 1), coupled with developing a microfluidic device with effective nano-stir bar mixing for improved detection efficiency (Aim 2), and then evaluate and validate the developed “liquid biopsy” system by multiplexed measuring serum Aβ40, Aβ42, tTau, pTau proteins and APP669-711 levels in AD patients longitudinally, and comparing and correlating the serum measurement with results from Aβ and tau PET imaging and proteomics analysis done on the same patients (Aim 3).

项目摘要阿尔茨海默氏病（AD）是主要的医疗保健和社会经济伯恩。和 有效的治疗尚待开发，在前驱阶段的早期检测和监视 进展是控制和干预AD开发的最佳方法。而非侵入性正电子 发射断层扫描（PET）可以测量大脑中的淀粉样蛋白β肽（AβS）和微管Tau蛋白 确认AD的发作，成像设备的可用性，单个Aβ或tau特异性的诊断准确性 放射性示例以及放射性材料的成本和暴露限制了早期筛选的成像应用 在监测疾病进程时，大量和潜水员的种群和反复的扫描。因此，有一个 未满足的需要开发可靠，易于访问且具有成本效益的体外诊断工具，以检测和 测量脑脊髓液中的AβS和TAU蛋白水平，甚至是患者或处于危险中的血液样本 个人处于“护理点”。但是，这些AD生物标志物的血清检测需要高灵敏度 由于血液样本中的AβS或TAU蛋白浓度低，并且慢速变化 这些生物标志物在疾病进展过程中的水平。对当前基于生物探测的一个主要挑战 体外诊断是生物污染效应，即，在表面上增加蛋白质和生物分子的吸附 检测代理和设备。这导致形成不需要的分子层，这会减少 通过（1）阻止对靶向生物标志物的抗体的检测特异性和灵敏度，以及（2）引起 吸附的非特异性蛋白质和其他分子的高“背景噪声”干扰了检测 有针对性的生物标志物。我们认为，我们的创新解决方案可以克服这些问题，即1） 抗生物性聚合物以覆盖纳米颗粒捕获剂以阻断非特异性蛋白吸附，从而阻止 保护靶向配体/抗体的灵敏度和特异性； 2）高磁力磁铁 氧化纳米棒（IONRS），具有纳米大小的磁性搅拌杆作用，以增强捕获剂和 微流体检测系统中的靶向生物标志物。通过整合这些技术，我们的项目目标 开发具有超高灵敏度和特异性的基于多路复用的“液体活检”系统 检测选定的AD生物标志物Aβ40，Aβ42，TOTAU（TTAU），磷酸化-TAU（PTAU）181和淀粉样蛋白 一个样品中的前体蛋白（APP）。我们将制造并优化抗双粘合离子 具有高效的流体混合和峰值Aβ40，Aβ42，TTAU的多路复用性检测的不同长宽比。 血样中的PTAU蛋白和App669-711（AIM 1），再加上与带有微流体设备的使用 有效的纳米刺激条混合以提高检测效率（AIM 2），然后评估和验证 通过多重测量血清Aβ40，Aβ42，TTAU，PTAU蛋白和 广告患者的APP669-711水平纵向，并将血清测量和相关 对同一患者进行的Aβ和TAU PET成像和蛋白质组学分析的结果（AIM 3）。