Photocleavage Technology for Blood-based Multi-Biomarker Alzheimer's Disease Assay

用于基于血液的多生物标志物阿尔茨海默病检测的光裂解技术

• 批准号: 10227129
• 负责人: Mark Lim
• 金额: $ 125万
• 依托单位: AMBERGEN, INC
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10227129
• 关键词: Abeta synthesis; Affect; Affinity; Alzheimer disease detection; Alzheimer' s Disease; Alzheimer' s disease diagnosis; Alzheimer' s disease diagnostic; Alzheimer' s disease model; Alzheimer' s disease patient; Alzheimer’s disease biomarker; Amyloid beta-42; Amyloid beta-Protein; Antibodies; Area Under Curve; Automation; Behavior; Benchmarking; Biological Assay; Biological Markers; Blood; Boston; Businesses; Calibration; Cerebrospinal Fluid Proteins; Chronic; Clinical; Cognitive; Cross Reactions; Custom; Dementia; Detection; Deterioration; Development; Diagnostic; Disease Progression; Early Diagnosis; Early Intervention; Evaluation; Gold; Health Care Costs; Immunoassay; Immunodiagnostics; Industry; Leadership; Licensing; Liquid substance; Longevity; Magnetism; Malignant neoplasm of prostate; Manufacturer Name; Measurement; Measures; Methods; Microspheres; Modeling; Neurodegenerative Disorders; Persons; Pharmacologic Substance; Phase; Phenotype; Population; Population Group; Positron-Emission Tomography; Public Health; ROC Curve; Reader; Reporting; Research; Research Personnel; Robotics; Role; Sampling; Seminal; Senile Plaques; Sepharose; Serology; Serum; Services; Specificity; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Spinal Puncture; Syndrome; Technology; Therapeutic; Universities; Work; aptamer; base; clinically relevant; cognitive function; commercialization; cost; design; detection limit; disorder control; human old age (65+); improved; malignant breast neoplasm; multiplex assay; multiplex detection; novel; novel strategies; particle; protein biomarkers; screening; tau Proteins

Summary/Abstract Alzheimer's Disease (AD) is a chronic neurodegenerative disease characterized by progressively worsening dementia, a syndrome associated with deterioration in cognitive function, behavior and ability to perform everyday activities. AD currently affects over 5.7 million persons in the U.S. with over 70% over the age of 65 and approximately 30 million world-wide. The public health costs due to AD is expected to grow exponentially due to the large expansion of the population over 65 which is estimated to reach over 100 million by 2060. Importantly, early diagnosis of AD is critical for application of therapeutics early in the disease progression, offering the promise of reducing debilitating effects and extending life span. Although the major clinical methods of diagnosing AD rely either on detection of biomarkers such as β-amyloid and tau proteins from cerebrospinal fluid (CSF) or PET imaging, these assays are limited by their invasiveness (e.g. lumbar puncture) or high costs, respectively, and not suitable as a front-line AD diagnostic. In contrast, a blood-based assay would be highly suitable for screening large population groups (e.g. persons over 60) for early AD. While blood-based assays of single biomarkers show significant promise for early detection of AD (e.g. for β-amyloid), it is likely that a panel of several biomarkers measured using a multiplex assay will ultimately emerge as necessary to achieve sufficient specificity to have clinical utility, especially in distinguishing different neurodegenerative diseases that have similar phenotypes. An additional major problem in the development of sensitive and accurate blood-based AD assays is the matrix effect. This effect is caused by the presence of the much more abundant molecules in blood which interfere with detection of the low-abundance biomarkers. We evaluated and successfully demonstrated during Phase I a new approach to multiplex serological AD assays termed PC-PURE™ which is designed to both enrich low-abundance biomarkers and eliminate the matrix effect. This technology is based on the use of novel photocleavable (PC) linkers developed by AmberGen which are incorporated into affinity capture agents such as aptamers or antibodies. We successfully demonstrated the feasibility of PC-PURE™ AD immunoassays by enrichment and multiplex detection of 3 model AD biomarkers (Aβ40, Aβ42 and tau). Our results show that PC-PURE™ when applied as a “front-end” to a commercially available multiplexed Luminex® xMAP® assay improves sensitivity by 30 to 667-fold compared to the same assay without PC-PURE™. Furthermore, PC-PURE™ provides a much more accurate estimate of absolute biomarker concentration since both the calibration curves and biomarkers extracted from the biospecimen are measured in a similar medium. During Phase II, PC-PURE™ will be further optimized for measurement of a model panel of 16 AD biomarkers using 300 AD confirmed and control serum samples and compared to results obtained using a bead- based immuno-MALDI-MS method that is currently one of the most accurate means of measuring AD biomarkers in serum. Commercial products will include PC-PURE™ multiplex immunoassay kits targeting AD biomarkers.

摘要/摘要 阿尔茨海默氏病（AD）是一种慢性神经退行性疾病，其特征是逐渐担心 痴呆症，一种与认知功能，行为和每天执行能力相关活性相关的综合征 活动。目前，广告在美国影响超过570万人，超过65岁，超过70％ 全球范围内约为3000万。由于AD引起的公共卫生费用预计将呈指数增长 到2060年，人口的大量扩大超过65岁，估计将超过1亿。 AD的早期诊断对于早期在疾病进展的早期治疗至关重要，提供了诺言 减少衰弱的效果并延长寿命。 尽管诊断AD的主要临床方法依赖于检测生物标志物，例如β-淀粉样蛋白和 来自脑脊液（CSF）或PET成像的Tau蛋白，这些测定受到其侵入性的限制（例如腰 穿刺）或高成本，不适合作为前线广告诊断。相反，基于血液的测定 非常适合筛选大量人群（例如60岁以上的人）。而基于血液 单个生物标志物的测定显示出对AD的早期检测（例如，对于β-淀粉样蛋白）的巨大希望，很可能是 使用多重测定测量的几种生物标志物的面板最终将出现以达到必要 具有临床实用性的足够特异性，尤其是在区分不同的神经退行性疾病时 具有相似的表型。敏感和准确的血液基于敏感和准确的血液的另一个主要问题 广告测定是矩阵效应。这种作用是由于血液中更丰富的分子的存在引起的 干扰了检测低丰度的生物标志物。 我们在第一阶段评估并成功证明了一种新的多重血清学广告测定方法 称为PC-Pure™，旨在富含低丰度生物标志物并消除基质效应。 该技术基于使用Ambergen开发的新型光电透明（PC）链接器的使用 掺入适体或抗体等亲和捕获剂中。我们成功证明了 PC-Pure™AD免疫测定的可行性通过富集和多重检测3模型AD生物标志物 （Aβ40，Aβ42和TAU）。我们的结果表明，将PC-Pure™应用于市售的“前端”时 多路复用Luminex®XMAP®分析与没有同一测定法相比，将灵敏度提高了30至667倍 PC-PURE™。此外，PC-Pure™提供了绝对生物标志物的更准确估计值 浓度是因为在A中测量了从生物测量中提取的校准曲线和生物标志物 相似的介质。在第二阶段，将进一步优化PC-Pure™，以测量16的模型面板 使用300 AD确认并控制血清样品的AD生物标志物，并将其与使用珠的结果进行了比较 基于Immuno-Maldi-MS方法目前是测量AD生物标志物的最准确方法之一 在血清中。商业产品将包括针对AD生物标志物的PC-Pure™多重免疫测定套件。