Flex-Hinge Bispecific Antibodies for Ultra-Sensitive Detection of Beta Amyloid Species in Plasma

用于超灵敏检测血浆中 β 淀粉样蛋白种类的 Flex-Hinge 双特异性抗体

• 批准号: 9408605
• 负责人: Daniel J. Capon
• 金额: $ 22.47万
• 依托单位: HINGE BIO, INC.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2018-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9408605
• 关键词: Affect; Affinity; Age-Years; Alzheimer' s Disease; American; Amyloid; Amyloid beta-Protein; Antibodies; Binding; Biological Assay; Biological Markers; Bispecific Antibodies; Blood; Blood Tests; Brain; Brain imaging; Brain scan; Cardiovascular Diseases; Cerebrospinal Fluid; Chemicals; Cholesterol; Clinical; Clinical Research; Clinical Trials Design; Detection; Development; Diagnosis; Diagnostic; Disease; Disease Progression; Fab Immunoglobulins; Family; Fc domain; Generations; Goals; Grant; Hand; Image; Impaired cognition; Impairment; Length; Ligation; Link; MALDI-TOF Mass Spectrometry; Mass Spectrum Analysis; Methods; Monitor; N-terminal; Neurodegenerative Disorders; Onset of illness; Pathologic; Patients; Peptides; Phase; Plasma; Positron-Emission Tomography; Procedures; Protein Engineering; Proteins; Proxy; Reproducibility; Resolution; Sampling; Sampling Studies; Sensitivity and Specificity; Site; Small Business Innovation Research Grant; Specificity; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Spinal Tap; Synapses; Technology; Testing; Therapeutic; aging population; antigen binding; base; cost; diagnostic assay; disease diagnosis; flexibility; improved; innovation; neuroimaging; novel; novel therapeutics; phase 2 study; screening; tau Proteins; validation studies

PROJECT SUMMARY/ABSTRACT Alzheimer’s disease (AD) is a devastating, progressive neurodegenerative disease affecting an estimated 11% of Americans over 65 years of age and 33-50% over 85 years of age. The initial pathological sign of AD, the build-up of beta amyloid (Aβ) plaques in the brain, often precedes impairment of cognition. Although these plaques can be imaged by Aβ PET brain scans or detected via analysis of Aβ peptides in the cerebrospinal fluid, these methods are costly, invasive and inconvenient. Recently, with clinical studies indicating that therapeutics may be more effective in treating AD at the earliest stages of the disease and that anti-Aβ therapy can remove Aβ from the brain, the need for a simple, inexpensive diagnostic blood test for AD onset and progression is even greater. It has been difficult to develop a blood test based on Aβ, in part because of the low concentration of Aβ peptides in blood and the high concentration of other proteins. We have approached this protein design problem by developing methods to link the antigen binding domains (Fabs) of antibodies to beads using flexible and extendible non-peptidyl hinges. The flexibly-linked Fabs on the beads are able to efficiently capture exceptionally low levels of Aβ (307 attomoles) in blood by binding cooperatively in a two-handed fashion. These Flex-Hinge™ Fab beads, combined with MALDI-TOF mass spectrometry, have revealed previously unknown forms of Aβ including a novel Aβ biomarker in plasma that acts as a precise qualitative and quantitative proxy for brain Aβ PET imaging, correlating with the stages of AD. In this Phase I SBIR, we shall improve upon these results in two key ways: (1) instead of using Flex-Hinge™ Fab beads, we shall produce Flex-Hinge™ Bispecific Antibody beads that we expect to have even greater sensitivity and utility through improved cooperativity and reduced variability, and (2) we shall move to Orbitrap technology, which has greater sensitivity and resolution for Aβ peptides compared to MALDI. We shall use these Flex-Hinge™ Bispecific Antibody beads and Orbitrap technology to examine clinical samples from the Swedish BioFINDER study to confirm the utility of the novel Aβ biomarker and identify additional Aβ biomarker(s) in plasma. Phase II studies shall evaluate this assay with larger sample sets. Ultimately, we intend to turn our test into a kit that will be broadly available and become routine for monitoring AD, much as cholesterol testing is used for monitoring cardiovascular disease.

项目总结/摘要 阿尔茨海默氏病（AD）是一种破坏性的、进行性的神经退行性疾病， 65岁以上的美国人和85岁以上的33-50%。AD最初的病理学症状是β-淀粉样蛋白的积聚 大脑中的淀粉样蛋白（Aβ）斑块通常先于认知障碍。虽然这些斑块可以通过Aβ成像 PET脑扫描或通过分析脑脊液中的Aβ肽来检测，这些方法是昂贵的，侵入性的， 不方便最近，临床研究表明，治疗剂在治疗AD时可能更有效， 疾病的最早阶段，抗A β治疗可以从大脑中清除Aβ，需要一种简单，廉价的 诊断性血液测试对于AD发作和进展的作用甚至更大。很难开发出一种基于 Aβ，部分原因是血液中Aβ肽浓度低，其他蛋白质浓度高。我们 已经通过开发方法来连接蛋白质的抗原结合结构域（Fab）来解决该蛋白质设计问题。 使用柔性和可延伸的非肽基铰链将抗体连接到珠。珠上的柔性连接的Fab能够 通过在一个双手的结合中协同作用，有效地捕获血液中异常低水平的Aβ（307阿托摩尔） 时尚.这些Flex-Hinge™ Fab珠与MALDI-TOF质谱法相结合，先前已经揭示了 未知形式的Aβ，包括血浆中的一种新型Aβ生物标志物，可作为精确的定性和定量指标 脑Aβ PET显像与AD分期相关。在第一阶段SBIR中，我们将在这些结果的基础上进行改进， 两个关键方法：（1）我们将生产Flex-Hinge™双特异性抗体珠粒，而不是使用Flex-Hinge™ Fab珠粒 我们期望通过提高协同性和减少可变性来获得更高的灵敏度和实用性，以及（2） 我们将转向Orbitrap技术，与MALDI相比，该技术对Aβ肽具有更高的灵敏度和分辨率。 我们将使用这些Flex-Hinge™双特异性抗体微珠和Orbitrap技术检查来自 瑞典BioFINDER研究，以确认新型Aβ生物标志物的效用，并确定其他Aβ生物标志物， 等离子体II期研究应使用较大的样本集评价该测定。最终，我们打算把我们的测试变成一个工具包， 这将是广泛可用的，并成为常规监测AD，就像胆固醇测试用于监测 心血管疾病