Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, China.
Analyst. 2013 May 7;138(9):2613-9. doi: 10.1039/c3an36744d.
In this work, we demonstrate the immunocapture and on-line fluorescence immunoassay of protein and virus based on porous polymer monoliths (PPM) in microfluidic devices. Poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) [poly(GMA-co-EGDMA)] monoliths were successfully synthesized in the polydimethylsiloxane (PDMS) microfluidic channels by in situ UV-initiated free radical polymerization. After surface modification, PPM provides a high-surface area and specific affinity 3D substrate for immunoassays. Combining with well controlled microfluidic devices, the direct immunoassay of IgG and sandwich immunoassay of inactivated H1N1 influenza virus using 5 μL sample has been accomplished, with detection limits of 4 ng mL(-1) and less than 10 pg mL(-1), respectively. The enhanced detection sensitivity is due to both high surface area of PPM and flow-through design. The detection time was obviously decreased mainly due to the shortened diffusion distance and improved convective mass transfer inside the monolith, which accelerates the reaction kinetics between antigen and antibody. This work provides a novel microfluidic immunoassay platform with high efficiency thereby enabling fast and sensitive immunoassay.
在这项工作中，我们展示了基于多孔聚合物整体材料（PPM）在微流控装置中对蛋白质和病毒的免疫捕获和在线荧光免疫分析。通过原位紫外引发自由基聚合，成功地在聚二甲基硅氧烷（PDMS）微流道中合成了聚（甲基丙烯酸缩水甘油酯-co-乙二醇二甲基丙烯酸酯）[聚（GMA-co-EGDMA）]整体材料。经过表面修饰，PPM 为免疫分析提供了高表面积和特定亲和力的 3D 基质。结合良好控制的微流控装置，使用 5 μL 样品完成了 IgG 的直接免疫分析和灭活 H1N1 流感病毒的夹心免疫分析，检测限分别为 4ng mL(-1)和小于 10pg mL(-1)。检测灵敏度的提高是由于 PPM 的高表面积和流动设计。检测时间明显缩短，主要是由于扩散距离缩短和整体内部对流质量传递的改善，从而加速了抗原和抗体之间的反应动力学。这项工作提供了一种具有高效率的新型微流控免疫分析平台，从而实现快速灵敏的免疫分析。
