Abstract

The immune response system of immunoassay microfluidic chips is a dynamic reaction process that continuously sends reactants to the surface of a solid carrier. Signal acquisition results from the heterogeneous immune reactions and reactant transport. Antibody immobilization is the most important part of heterogeneous immune reactions, and reactant transport is reflected in the form of fluid velocity. Here, we reported several surface modification processes on polystyrene substrates that are employed to study the relationship between the antibody immobilization and flow behavior in heterogeneous immune response processes. The antibody was immobilized using covalent grafting. Based on the mechanism of sandwich enzyme linked immunosorbent assay, a fluorescence quantitative detection method was used to evaluate the immune response process. The effects of different surface modification processes on immune response and flow behavior were studied. We identified an optimal flow velocity in the dynamic immune response system in the microfluidic chip. The immune response signal was the strongest when the average flow velocity was approximately 0.2 mm/s in the procalcitonin detection system. Compared with the amino and aldehyde group substrates, the epoxy group substrate has the highest antibody immobilization efficiency; compared with the surface modified by small molecular groups, the introduction of Poly-L-Lysine can increase the amount of antibody immobilization.

摘要

免疫微流控芯片的免疫反应系统是一个动态的反应过程，它不断地将反应物送到固体载体的表面。信号采集来自异质免疫反应和反应物转运。抗体固定化是异质免疫反应中最重要的部分，反应物转运以流体速度的形式体现。在这里，我们报告了聚苯乙烯基质上的几种表面修饰过程，这些过程用于研究异质免疫反应过程中抗体固定化和流动行为之间的关系。使用共价接枝固定抗体。基于夹心酶联免疫吸附测定的机理，采用荧光定量检测方法评价免疫反应过程。研究了不同表面修饰工艺对免疫反应和流动行为的影响。我们确定了微流控芯片中动态免疫反应系统的最佳流速。在降钙素原检测系统中，平均流速约为 0.2 mm/s 时免疫反应信号最强。与氨基和醛基底物相比，环氧基底物的抗体固定效率最高；与小分子基团修饰的表面相比，Poly-L-Lysine的引入可以增加抗体的固定量。在降钙素原检测系统中，平均流速约为 0.2 mm/s 时免疫反应信号最强。与氨基和醛基底物相比，环氧基底物的抗体固定效率最高；与小分子基团修饰的表面相比，Poly-L-Lysine的引入可以增加抗体的固定量。在降钙素原检测系统中，平均流速约为 0.2 mm/s 时免疫反应信号最强。与氨基和醛基底物相比，环氧基底物的抗体固定效率最高；与小分子基团修饰的表面相比，Poly-L-Lysine的引入可以增加抗体的固定量。