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

护理点（PoC）需要高度敏感的分析，以实现快速诊断和及时治疗。较短的检测时间尤为重要。然而，在PoC条件下（小型紧凑设备，可变环境条件）提供经济合理且高度敏感的分析是极具挑战性的。离心微流体有可能自动进行小分析物浓度（< 10pg/ml）的PoC测试。离心重力场中可伸缩的体积力使流体操作实现强大的自动化，广泛独立于可变的样品基质（粘度和润湿特性）和环境条件（温度，压力，湿度等）。所有重要的过程都是由转子的旋转频率控制的。测试芯片的接口很简单，因为不需要油管或泵。PoC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。为了缩短分析时间并达到高灵敏度，分析物向功能化固相的传递和非仿射物质的去除必须非常高效。然而，目前的技术状况并没有提供离心微流体中输运现象的基本知识。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分析物向固相的最终输运的影响仍未得到解决。因此，到目前为止，基于离心微流体的高灵敏度PoC测试尚未得到证实。本项目的目标是建立离心式微流控输运现象和亲和反应中分析物输运的基本知识。应评估不同的环境条件和样品基质对结合效率和速度的影响。基于这方面的知识，高灵敏度的PoC测试将与离心微流体自动化。本项目旨在实现以下子目标：建立离心微流控反应室中分析物和分析物-共轭物-配合物的对流和扩散输运模型以及分析物与固相结合模型。开发模型验证实验。通过实施高灵敏度的肌钙蛋白测定（检测限< 3pg/ml；在10pg /ml时<10% CV）来演示基于模拟的布局。临床样品验证。建立了离心微流控PoC试验设计组件库。

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.