Holladay James D, Berkheimer Zachary A, Haggard Michael K, Nielsen Jacob B, Nordin Gregory P, Woolley Adam T
Department of Chemistry and Biochemistry, Brigham Young University, Provo, Utah 84602, United States.
Department of Electrical and Computer Engineering, Brigham Young University, Provo, Utah 84602, United States.
Precis Chem. 2025 Mar 3;3(5):261-271. doi: 10.1021/prechem.4c00092. eCollection 2025 May 26.
A miniaturized, biomarker-based diagnostic for preterm birth (PTB) risk will require multiple sample preparation steps to be integrated in a single platform. To this end, we created a 3D printed microfluidic device that combines immunoaffinity extraction (IAE), solid-phase extraction (SPE), and fluorescent labeling. This device uses an antibody-functionalized IAE monolith to selectively extract PTB biomarkers, a lauryl methacrylate reverse-phase SPE monolith to concentrate and facilitate fluorescent labeling of PTB biomarkers, and 3D printed valves to control flow through the monoliths. The advantageous iterative design process for arriving at a functional device is documented. The IAE/SPE device performed selective, reproducible extractions of three PTB biomarkers from buffer and depleted maternal blood serum, demonstrating its utility for single-biomarker and multiplexed extractions. After tandem extraction and fluorescent labeling, biomarkers eluted from the SPE monolith in a concentrated plug, facilitating future integration with downstream analysis techniques including microchip electrophoresis. This device effectively combines and automates orthogonal chromatographic extraction methods and constitutes a substantial step toward a complete microfluidic PTB prediction platform.
一种基于生物标志物的早产（PTB）风险小型化诊断方法需要将多个样品制备步骤集成在一个单一平台上。为此，我们制造了一种3D打印微流控装置，该装置结合了免疫亲和萃取（IAE）、固相萃取（SPE）和荧光标记。该装置使用抗体功能化的IAE整体柱选择性萃取PTB生物标志物，月桂基甲基丙烯酸酯反相SPE整体柱浓缩并促进PTB生物标志物的荧光标记，以及3D打印阀门来控制通过整体柱的流动。记录了获得功能装置的有利迭代设计过程。IAE/SPE装置从缓冲液和耗尽的母体血清中对三种PTB生物标志物进行了选择性、可重复的萃取，证明了其在单生物标志物和多重萃取中的效用。经过串联萃取和荧光标记后，生物标志物以浓缩塞的形式从SPE整体柱中洗脱出来，便于未来与包括微芯片电泳在内的下游分析技术整合。该装置有效地结合并自动化了正交色谱萃取方法，朝着完整的微流控PTB预测平台迈出了重要一步。
