Multiplexed immunoassay for building patient-specific molecular profiles of CSF amyloid beta and TAU

用于构建患者特异性 CSF 淀粉样蛋白 β 和 TAU 分子谱的多重免疫分析

• 批准号: 10707200
• 负责人: Vladislav Bergo
• 金额: $ 143.37万
• 依托单位: ADEPTRIX CORPORATION
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10707200
• 关键词: Age; Alzheimer' s Disease; Alzheimer' s disease related dementia; Alzheimer’s disease biomarker; Amino Acid Sequence; Amyloid beta-Protein; Biological Assay; C-terminal; Cerebrospinal Fluid; Clinical; Clip; Cognitive; Complement; Complex; Computer software; Data; Digestion; Disease; Disease Progression; Ensure; Epitopes; Gender; Human; Immunoassay; Individual; Life; Light; Mass Spectrum Analysis; Measures; Methods; Modification; Molecular Profiling; N-terminal; Nerve Degeneration; Neurodegenerative Disorders; Patients; Peptides; Performance; Phase; Phosphorylation Site; Plasma; Protein Isoforms; Proteins; Proteomics; Pyroglutamate; Race; Reagent; Recovery; Reporting; Research Personnel; Sampling; Sensitivity and Specificity; Serum; Source; Specificity; Variant; Work; abeta accumulation; accurate diagnostics; alpha synuclein; antibody conjugate; biobank; combinatorial; detection limit; diagnostic signature; drug discovery; improved; industry partner; mild cognitive impairment; neurofilament; novel; novel marker; polypeptide; potential biomarker; product development; profiles in patients; prognostic signature; progression marker; protein biomarkers; screening; specific biomarkers; synthetic peptide; tau Proteins; tool

ABSTRACT This project will develop a multiplexed immunoassay for measuring patient-specific molecular profiles of the two protein markers of Alzheimer’s disease (AD): amyloid beta (ABeta) and TAU from human cerebrospinal fluid (CSF). Both proteins are structurally complex due to the existence of multiple differentially cleaved forms (ABeta) and 6 isoforms containing 50+ phosphorylation sites (TAU). CSF provides a rich source of potential biomarkers, which can be extensively mined to build diagnostic and prognostic signatures of AD and AD-related dementias (ADRD) or to develop personalized patient profiles for drug discovery. At present, those efforts are constrained by the lack of technical capability to resolve multiple ABeta and TAU proteoforms, more specifically by the lack of products for multiplexed capture and enrichment of ABeta and TAU for quantitative mass spectrometry-based proteomics. This proposal seeks to build upon an earlier successful product development project performed by Adeptrix for a pharma customer. Using our BAMS™ platform, we were able to identify at least 14 novel low abundance CSF ABeta peptides and dramatically expand the sequence coverage of CSF TAU by adding two new types of probes, which complement the existing conventional probes for total (tTAU) and phospho-TAU (pTAU). The proposed immunoassay, termed ABeta/TauScan™ will address the current need of biologists, clinicians, and pharma for comprehensive molecular profiling tools for studying protein markers of neurodegeneration. Furthermore, it will open a path to developing similar assays for other protein targets, such as neurofilament light polypeptide (NFL) and alpha-synuclein. While ABeta/TauScan™ is intended primarily for CSF, post Phase II the reagents will be evaluated for use in serum/plasma. Throughout this project we will work closely with academic, clinical and industry partners to ensure the robust analytical performance of ABeta/TauScan™. Once the assay is created, it will be validated by screening 300 CSF samples representing cognitively normal (CN) subjects, mild cognitive impairment (MCI) and AD subjects and establishing a molecular signature of CN to MCI to AD progression that will contain at least one novel marker for improved assay sensitivity and specificity.

抽象的 该项目将开发一种多重免疫测定法，用于测量患者特异性的两种分子特征 阿尔茨海默病 (AD) 的蛋白质标志物：来自人脑脊液 (CSF) 的β淀粉样蛋白 (ABeta) 和 TAU。 由于存在多种差异切割形式 (ABeta) 和 6，两种蛋白质的结构都很复杂 含有 50 多个磷酸化位点 (TAU) 的同工型。 CSF 提供了丰富的潜在生物标志物来源， 可以广泛挖掘以建立 AD 和 AD 相关痴呆症 (ADRD) 的诊断和预后特征 或为药物发现开发个性化的患者档案。目前，这些努力受到以下因素的限制： 缺乏解决多种 Aβ 和 TAU 蛋白异构体的技术能力，更具体地说是缺乏产品 用于 Aβ 和 TAU 的多重捕获和富集，用于基于定量质谱的蛋白质组学。 该提案旨在以 Adeptrix 早期成功的产品开发项目为基础 一位制药客户。使用我们的 BAMS™ 平台，我们能够识别至少 14 种新型低丰度 CSF Aβ 肽并通过添加两种新型探针显着扩大 CSF TAU 的序列覆盖范围， 它补充了现有的总 TAU (tTAU) 和磷酸化 TAU (pTAU) 常规探针。拟议的 称为 Abeta/TauScan™ 的免疫测定将满足生物学家、临床医生和制药公司当前的需求 用于研究神经变性蛋白质标记物的综合分子分析工具。此外，它将 为开发其他蛋白质靶点的类似检测方法开辟了道路，例如神经丝轻链多肽 (NFL) 和α-突触核蛋白。虽然 Abeta/TauScan™ 主要用于 CSF，但在 II 期后将对试剂进行评估 用于血清/血浆。 在整个项目中，我们将与学术、临床和行业合作伙伴密切合作，以确保稳健的 ABeta/TauScan™ 的分析性能。创建检测后，将通过筛选 300 个脑脊液进行验证 代表认知正常 (CN) 受试者、轻度认知障碍 (MCI) 和 AD 受试者的样本 建立 CN 到 MCI 到 AD 进展的分子特征，其中至少包含一个新的标记物 提高检测灵敏度和特异性。