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

需要对护理点(PoC)进行高度敏感的分析，以实现快速诊断和及时治疗。短的化验时间尤其重要。然而，要在PoC条件下(小型紧凑型设备，可变环境条件)提供经济合理和高灵敏度的分析是极其具有挑战性的。离心式微流控有可能实现对小浓度分析物(&lt；10pg/ml)的PoC测试的自动化。离心重力场中的可伸缩体积力可实现流体操作的强大自动化，完全独立于不同的样品矩阵(粘度和润湿特性)和环境条件(温度、压力、湿度等)。所有重要的过程都只受转子的旋转频率控制。测试芯片的接口很简单，因为不需要管子或泵。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，以便随后进行定量。为了保持较短的分析时间并达到高灵敏度，分析物向功能化固相的传输和非仿生物质的去除必须非常有效。然而，目前的技术水平并没有提供离心式微流控中传输现象的基本知识。惯性力(离心力、欧拉力和科里奥利力)的相互作用对流型的影响以及由此导致的分析物向固相的传输仍未得到解决。因此，基于离心式微流体的高灵敏度POC测试到目前为止还没有得到证实。本项目的目标是为亲和反应建立离心式微流控传输现象和分析物传输的基础知识。应评估环境条件和样品基质变化对结合效率和速度的影响。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结合络合物的对流和扩散传输以及分析物与固相的结合模型。开发用于模型验证的实验。通过实现高灵敏的肌钙蛋白分析(检测下限为3pg/ml；&lt；10%变异系数为10pg/ml)来演示基于模拟的布局。通过临床样本验证。建立了用于设计离心式微流控POC测试的组件库。

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.