Integrated microfluidic devices for preterm birth biomarker measurement

用于早产生物标志物测量的集成微流体装置

• 批准号: 8442695
• 负责人: Adam Thomas Woolley
• 金额: $ 33.14万
• 依托单位: BRIGHAM YOUNG UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8442695
• 关键词: Address; Adsorption; Affinity Chromatography; Biological Assay; Biological Markers; Birth; Blinded; Blood; Blood specimen; Caring; Coupling; Defensins; Development; Device or Instrument Development; Devices; Diagnosis; Evaluation; Excision; Fingers; Fluorescence; Goals; Health; Healthcare; Hour; Human; Human Resources; Immunoassay; Individual; Label; Lactoferrin; Measurement; Measures; Medical; Membrane; Methods; Microchip Electrophoresis; Microfluidic Microchips; Microfluidics; Nature; Neonatal; Newborn Infant; Peptides; Performance; Phase; Polymers; Pregnancy Complications; Pregnant Women; Premature Birth; Preparation; Prevalence; Process; Proteins; Pump; Reagent; Risk; Risk Assessment; Sampling; Serum; Severities; Solid; Specificity; System; Testing; Therapeutic Intervention; United States; Universities; Utah; Work; design; improved; instrument; medical schools; meetings; microchip; neonatal death; operation; pressure; prevent; public health relevance; research study; tool

DESCRIPTION (provided by applicant): Preterm birth (PTB) is the most common complication of pregnancy, with over 500,000 early births annually in the United States. PTB is responsible for the majority of neonatal deaths and newborn illnesses. Unfortunately, the ability to assess PTB risk prior to contractions is not available clinically; this capability would enable therapeutic interventions that prevent or forestall delivery, potentially decreasing PTBs and/or the severity of neonatal complications. This renewal proposal focuses on the development of an integrated microfluidic platform for rapid, sensitive and inexpensive quantitation of serum PTB risk biomarkers (newly discovered peptides and known proteins), weeks before contractions occur. This proposal thus offers a major human health impact, potentially decreasing the prevalence of PTBs and their associated complications. This proposal tests the hypothesis that further development of microfluidic systems, which combine sample preparation, separation and quantitation in a single platform, will enable the direct measurement of PTB biomarkers in serum, allowing risk assessment and preventative measures to occur well before contractions begin. The proposed automated microfluidic platform has three main components: (1) a multiplexed immunoaffinity extraction module to selectively capture unlabeled PTB biomarkers; (2) a purification/labeling module to enrich PTB biomarkers and then allow fluorescence labeling with removal of unreacted tag; and (3) a microchip electrophoresis module to separate the labeled PTB biomarkers, enabling their quantitation. This approach improves significantly over present instrument- and personnel-intensive methods to determine PTB biomarkers, and should offer specificity comparable to sandwich immunoassays that would be problematic to develop for small peptide targets. The goal of this renewal proposal is to further develop integrated microfluidic systems for convenient, inexpensive, rapid and quantitative analysis of serum biomarkers with PTB risk correlation. This objective will be accomplished via three specific aims. In Aim 1 electrokinetically operated microchips to quantify PTB biomarkers will be created and tested, integrating immunoaffinity extraction, solid-phase enrichment, fluorescence labeling, electrophoretic separation and biomarker quantitation. Aim 2 will involve developing PTB biomarker analysis microdevices that use pressure actuation of on-chip valves and pumps to carry out integrated sample preparation operations. Performance of the devices from Aims 1 and 2 will then be compared. In Aim 3 these microfluidic platforms will be used to measure PTB biomarkers in blood samples to establish the feasibility of microchip methods in predicting PTB risk weeks before contractions occur. Importantly, this work addresses the urgent need to diagnose PTB risk when therapeutic intervention is still feasible; however, the general nature of the approach should be applicable well beyond biomarkers for PTB, further exemplifying the large and important health impact of these studies.

描述（由申请人提供）：早产 (PTB) 是最常见的妊娠并发症，美国每年有超过 500,000 例早产。 PTB 是导致大多数新生儿死亡和新生儿疾病的原因。不幸的是，临床上尚无法在宫缩前评估 PTB 风险。这种能力将使预防或阻止分娩的治疗干预成为可能，从而有可能减少 PTB 和/或新生儿并发症的严重程度。该更新提案的重点是开发一个集成的微流体平台，用于在宫缩发生前几周快速、灵敏且廉价地对血清 PTB 风险生物标志物（新发现的肽和已知蛋白质）进行定量。因此，该提案对人类健康产生了重大影响，有可能降低 PTB 及其相关并发症的患病率。 该提案测试了这样的假设：微流体系统的进一步发展将样品制备、分离和定量结合在一个平台中，将能够直接测量血清中的 PTB 生物标志物，从而在宫缩开始之前就可以进行风险评估和预防措施。所提出的自动化微流控平台具有三个主要组成部分：（1）多重免疫亲和提取模块，用于选择性捕获未标记的 PTB 生物标志物； (2) 纯化/标记模块，用于富集 PTB 生物标志物，然后进行荧光标记并去除未反应的标签； (3) 微芯片电泳模块，用于分离标记的 PTB 生物标志物，并对其进行定量。与目前测定 PTB 生物标志物的仪器和人员密集型方法相比，这种方法显着改进，并且应该提供与夹心免疫测定相当的特异性，而夹心免疫测定在开发小肽靶标时会存在问题。 该更新提案的目标是进一步开发集成微流体系统，以方便、廉价、快速和定量地分析与 PTB 风险相关的血清生物标志物。这一目标将通过三个具体目标来实现。在目标 1 中，将创建和测试用于量化 PTB 生物标志物的电动操作微芯片，集成免疫亲和提取、固相富集、荧光标记、电泳分离和生物标志物定量。目标 2 将涉及开发 PTB 生物标志物分析微型设备，该设备使用片上阀门和泵的压力驱动来执行集成的样品制备操作。然后将比较目标 1 和 2 的设备的性能。在目标 3 中，这些微流体平台将用于测量血液样本中的 PTB 生物标志物，以确定微芯片方法在宫缩发生前几周预测 PTB 风险的可行性。 重要的是，这项工作解决了在治疗干预仍然可行时诊断 PTB 风险的迫切需要；然而，该方法的一般性质应该远远超出 PTB 生物标志物的适用范围，进一步证明了这些研究对健康的巨大而重要的影响。