Lagrangian detection of biomolecular interactions

生物分子相互作用的拉格朗日检测

• 批准号: 6869526
• 负责人: Frederick R Haselton
• 金额: $ 18.88万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6869526
• 关键词: automated data processing; intermolecular interaction; nanotechnology; nucleic acid hybridization; nucleic acid probes; nucleic acid purification; nucleic acid quantitation /detection; polymerase chain reaction; surface property; technology /technique development; automated data processing; intermolecular interaction; nanotechnology; nucleic acid hybridization; nucleic acid probes; nucleic acid purification; nucleic acid quantitation /detection; polymerase chain reaction; surface property; technology /technique development

DESCRIPTION (provided by applicant): Patterns of gene expression and protein profiles are currently revolutionizing the understanding of biology and disease and are likely to become diagnostic and prognostic tools. Once these patterns are established, methods to implement them will be required. We have designed a platform which rapidly matches the characteristics of an unknown sample with established expression or protein profiles. The approach is characterized by computer controlled testing for pattern features with adaptive feedback to increase sensitivity and reliability. It will have greatest utility for identifying established patterns described in terms of the presence or concentration of 100s to 1000s of key molecular structures. Current high-throughput methods for profiling of an unknown sample, such as DNA microarrays for gene expression profiling, can be characterized as Eulerian design - known capture probes are fixed in space by attachment to a solid substrate and hybridized by target diffusion. Biorecognition is inefficient querying only a fraction of the target solution, target-probe interactions occur slowly, and target-probe binding occurs under uniform conditions. This application describes an alternative Lagrangian design - probes are arranged on a filament and moved through nanoliter reaction zones for biorecognition testing, processing, and analysis. The extremely small dimensions enable complete, rapid, and complete target-probe interactions. The flexible and adaptive features permit tailoring reaction conditions to each capture probe-target interaction. We have identified several key components that are critical to the success of this new platform technology. These are the identification of an appropriate fiber, deposition of probe, development of the small volume reaction compartment with immiscible surface tension valves, and a specialized apparatus for computer controlled fiber transport through multiple reaction chambers. Although we have limited experimental data, the data we do have, suggests that each of the key components of the proposed system is feasible. This technology will provide a new tool for the identification of patterns present in gene expression or protein profiles enabling these profiles to be fully utilized for diagnostic and prognostic purposes.

描述（由申请人提供）：基因表达和蛋白质谱的模式目前正在彻底改变对生物学和疾病的理解，并可能成为诊断和预后工具。一旦建立了这些模式，就需要实施这些模式。我们设计了一个平台，该平台将未知样品的特征与已建立的表达或蛋白质曲线匹配。该方法的特征是对具有自适应反馈的模式特征的计算机控制测试，以提高灵敏度和可靠性。它将具有最大的效用，以识别根据100s至1000s关键分子结构的存在或浓度所描述的已建立模式。 当前用于分析未知样品的高通量方法，例如用于基因表达分析的DNA微阵列，可以表征为欧拉设计 - 已知的捕获探针通过附着在固体基板上并通过靶标扩散杂交。生物识别仅在目标溶液中的一小部分，目标 - 探针相互作用发生缓慢，而目标探针结合在均匀条件下发生。该应用程序描述了另一种拉格朗日设计 - 探针被排列在细丝上，并通过纳米素反应区进行了生物识别测试，加工和分析。极小的尺寸可以实现完整，快速和完整的目标探针相互作用。柔性和适应性特征允许为每个捕获探针目标相互作用定制反应条件。 我们已经确定了几个关键组件，这些组件对于这项新平台技术的成功至关重要。这些是适当的纤维，探针沉积，与不混溶的表面张力阀的小体积反应的发展以及通过多个反应室进行计算机控制的纤维传输的专门设备。尽管我们的实验数据有限，但我们确实拥有的数据表明，所提出系统的每个关键组成部分都是可行的。 该技术将为识别基因表达或蛋白质曲线中存在的模式提供一个新工具，从而使这些概况充分利用用于诊断和预后。