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: 10602768
• 负责人: Tamil Selvan Anthonymuthu
• 金额: $ 104万
• 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 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10602768
• 关键词: Address; 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; Site; Source; Specificity; Variant; Work; abeta accumulation; accurate diagnostics; alpha synuclein; antibody conjugate; base; 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; tau-1; 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 该项目将开发多重免疫测定法，用于测量两者的患者特异性分子谱 阿尔茨海默氏病（AD）的蛋白质标志物：淀粉样蛋白β（ABETA）和人脑脊髓液（CSF）的蛋白质标志物。 由于存在多种不同的分裂形式（ABETA）和6 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， 可以广泛开采以建立AD和AD相关痴呆症（ADRD）的诊断和预后特征 或开发个性化的患者概况以发现药物。目前，这些努力受到 缺乏解决多个Abeta和Tau蛋白质成型的技术能力，更具体地说是由于缺乏产品 用于多重捕获和富集Abeta和Tau，以用于定量质谱蛋白质组学。 该提案旨在以Adeptrix执行的早期成功的产品开发项目为基础 制药客户。使用我们的BAMS™平台，我们能够识别至少14个新颖的低丰度CSF Abeta肽和通过添加两种新类型的问题，大大扩展CSF TAU的序列覆盖范围： 完成总计（TTAU）和Phosho-Tau（PTAU）的现有常规问题。提议 免疫测定法，称为ABETA/TAUSCAN™将满足生物学家，临床医生和制药药的当前需求 用于研究神经退行性的蛋白质标志物的综合分子分析工具。此外，它将 打开开发其他蛋白质靶标类似测定的途径，例如神经丝光多肽（NFL） 和α-核蛋白。虽然Abeta/Tauscan™主要用于CSF，但第二阶段后的试剂将被评估 用于血清/血浆。 通过这个项目，我们将与学术，临床和行业合作伙伴紧密合作，以确保强大 Abeta/Tauscan™的分析性能。一旦创建测定，它将通过筛选300 CSF进行验证 代表认知正常受试者，轻度认知障碍（MCI）和AD受试者的样本 建立CN与MCI的分子签名以进行AD进展，该进展至少包含一个新颖的标记 提高测定敏感性和特异性。