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: 8701291
• 负责人: Adam Thomas Woolley
• 金额: $ 28.87万
• 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/8701291
• 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）是最常见的怀孕并发症，每年在美国有超过500,000个早期出生。 PTB负责大多数新生儿死亡和新生儿疾病。不幸的是，在临床上无法获得收缩之前评估PTB风险的能力；这种能力将使治疗性干预措施可防止或阻止递送，可能会降低PTB和/或新生儿并发症的严重程度。该更新建议着重于开发综合微流体平台，用于快速，敏感和廉价的血清PTB风险生物标志物（新发现的肽和已知蛋白质），在收缩发生前几周。因此，该提案提供了重大的人类健康影响，可能会降低PTB的患病率及其相关并发症。 该提案检验了以下假设：在单个平台中结合样品制备，分离和定量的微流体系统的进一步开发将使PTB生物标志物在血清中的直接测量，从而使风险评估和预防措施在宫缩开始前就可以很好地发生。提出的自动化微流体平台具有三个主要组成部分：（1）多重的免疫亲和力提取模块，可选择性地捕获未标记的PTB生物标记物； （2）纯化/标记模块以丰富PTB生物标志物，然后允许荧光标记并去除未反应的TAG； （3）一个微芯片电泳模块，以分离标记的PTB生物标志物，以实现其定量。这种方法在当前的仪器和人员密集型方法上可以显着改善，以确定PTB生物标志物，并且应提供与三明治免疫测定相当的特异性，而对于小肽靶标来说，这将是有问题的。 该更新建议的目的是进一步开发集成的微流体系统，以方便，廉价，快速和定量分析具有PTB风险相关性的血清生物标志物。该目标将通过三个特定目标来实现。在AIM 1中，将创建和测试用于量化PTB生物标志物的电动操作的微芯片，从而整合免疫亲和力提取，固体富集，荧光标记，电泳分离和生物标志物定量。 AIM 2将涉及开发PTB生物标志物分析的微型电视，以使用芯片阀和泵的压力致动以进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AIM 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预测收缩前几周的PTB风险中的可行性。 重要的是，这项工作解决了治疗干预仍然可行的迫切需要诊断PTB风险；但是，该方法的一般性质应适用于PTB的生物标志物，进一步体现了这些研究的巨大健康影响。