Simulation based characterization of transport phenomena and affinity reactions at the solid phase in centrifugal microfluidics

基于模拟的离心微流体中固相传输现象和亲和反应的表征

• 批准号: 286516335
• 负责人: Privatdozent Dr.-Ing. Nils Paust
• 金额: --
• 依托单位: Professur für Anwendungsentwicklung
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2015
• 资助国家: 德国
• 起止时间: 2014-12-31 至 2019-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/286516335?language=en
• 关键词: Simulation; based; characterization; transport; phenomena

Highly sensitive analysis is required for the point of care (PoC) to enable rapid diagnostics and timely treatment. A short assay time is in particular important. However, to provide economically reasonable and highly sensitive analysis under PoC conditions (small compact devices, variable ambient conditions) is extremely challenging.Centrifugal microfluidics has the potential to automate PoC tests for small analyte concentrations (< 10pg/ml). Scalable volume forces in the centrifugal gravity field enable robust automation of fluidic operations, widely independent from variable sample matrixes (viscosity and wetting properties) and ambient conditions (temperature, pressure, humidity, etc.). All important processes are controlled by the rotational frequency of the rotor, only. Interfaces of the testchips are simple because no tubing or pumps are required.PoC tests (immuno- and molecular diagnostic) are typically based on affinity reactions that separate the analyte from the sample matrix for subsequent quantification. To keep the analysis time small and reach high sensitivity, the transport of analyte to the functionalized solid phase and the removal of non-affine substances has to be very efficient. However, the state of the art does not provide fundamental knowledge of transport phenomena in centrifugal microfluidics. The impact of the interplay of inertia forces (centrifugal-, Euler- and Coriolis force) on flow profiles and resulting transport of analyte to the solid phase remains unresolved. Consequently, highly sensitive PoC tests based on centrifugal microfluidics have not been demonstrated so far. The goal of this project is to establish fundamental knowledge of centrifugal microfluidic transport phenomena and analyte transport for affinity reactions. The impact of varying ambient conditions and sample matrixes on binding efficiency and speed should be evaluated. Based on this knowledge highly sensitive PoC tests will be automated with centrifugal microfluidics.This project aims to achieve the following sub-goals:Establish models of convective and diffusive transport of analyte and analyte-conjugate-complexes and of binding of analytes to solid phases in centrifugal microfluidic reaction chambers.Develop experiments for model validation.Demonstration of simulation based layout by implementing a highly sensitive Troponin assay (detection limit < 3pg/ml; <10% CV at 10 pg/ml). Validation with clinical samples.Establish a component library for the design of centrifugal microfluidic PoC tests.

护理点（Point of Care，简称POS）需要高灵敏度的分析，以实现快速诊断和及时治疗。短的测定时间是特别重要的。然而，在微流控条件下（小型紧凑的设备，可变的环境条件）提供经济合理且高度灵敏的分析是极其具有挑战性的。离心微流控技术具有自动化微流控检测小分析物浓度（<10 pg/ml）的潜力。离心重力场中的可缩放体积力能够实现流体操作的强大自动化，广泛独立于可变样品基质（粘度和润湿特性）和环境条件（温度、压力、湿度等）。所有重要的过程仅由转子的旋转频率控制。测试芯片的接口很简单，因为不需要管道或泵。免疫学测试（免疫和分子诊断）通常基于亲和反应，将分析物从样品基质中分离出来，用于随后的定量。为了保持分析时间短并达到高灵敏度，分析物到功能化固相的传输和非仿射物质的去除必须非常有效。然而，现有技术没有提供离心微流体中的传输现象的基本知识。惯性力（离心力，欧拉力和科里奥利力）的相互作用对流动剖面和由此产生的分析物到固相的运输的影响仍然没有得到解决。因此，基于离心微流体的高灵敏度微流控测试迄今尚未得到证实。本项目的目标是建立离心微流体传输现象和亲和反应的分析物传输的基础知识。应评估不同环境条件和样品基质对结合效率和速度的影响。基于这一认识，高灵敏度的肌钙蛋白检测将通过离心微流控技术实现自动化。该项目旨在实现以下子目标：建立分析物和分析物-缀合物-复合物的对流和扩散传输模型以及分析物与离心微流控反应室中固相结合的模型。开发模型验证实验。通过实施高灵敏度肌钙蛋白检测（检测限<3 pg/ml;在10 pg/ml时<10%CV）来演示基于模拟的布局。临床样品验证：建立离心微流控电泳测试设计的元件库。