Department of Bionanotechnology, Gachon University, Seongnam 13120, Republic of Korea.
Philmedi R&D Center, Philmedi Incorporation, 33, Sagimakgol-ro 62beon-gil, Jungwon-gu, Seongnam 13211, Republic of Korea.
J Immunol Methods. 2024 Jul;530:113690. doi: 10.1016/j.jim.2024.113690. Epub 2024 May 15.
In general, vertical flow assay (VFA) has a disadvantage of requiring a complex analysis process that involves manually injecting various reagents (target analyte, washing buffer, detection conjugate, etc.) sequentially. However, in this study, we have developed an innovative paper-based VFA device that replaces the complex analysis process with one-step and enables the detection of multiple targets. The fabrication process of the multi-target detection VFA device is as follows: preparation and pre-treatment of the strip materials, design of strip cartridge, design of the multiple detection VFA device, optimization experiments for strip sample flow rates, determination of device analysis time, determination of device limit of detection (LOD), multiple target signal uniformity experiment, immunoglobulin G (IgG) and C-reactive protein (CRP) antigen-antibody multiple detection experiment, and data extraction and analysis method. The use of paper-based materials enables the device to be produced at cost-effective, and cartridge production allowed for uniform array formation. IgG and CRP are used to evaluate the performance of the device as common biomarkers. The device proposed in this study is currently under research. To validate multiple target detection capability of the VFA device proposed in this study, two types of antigens-antibodies (Human IgG and Human CRP) were employed. The detection limit is 0.15 μg/mL for IgG and 0.19 μg/mL for CRP in naked eye. Furthermore, it was confirmed that there is no cross-reactivity caused by the device structure through IgG and CRP antigens. In conclusion, the VFA device proposed in this study consists of a one-step analysis process, and it has been confirmed that it can detect multiple targets simultaneously.
一般来说，垂直流动分析（VFA）的缺点是需要复杂的分析过程，需要手动依次注入各种试剂（目标分析物、洗涤缓冲液、检测结合物等）。然而，在本研究中，我们开发了一种创新的基于纸张的 VFA 设备，该设备用一步分析过程取代了复杂的分析过程，并能够同时检测多个目标。多目标检测 VFA 设备的制造过程如下：条带材料的准备和预处理、条带盒的设计、多目标检测 VFA 设备的设计、条带样品流速的优化实验、设备分析时间的确定、设备检测限（LOD）的确定、多个目标信号均匀性实验、免疫球蛋白 G（IgG）和 C-反应蛋白（CRP）抗原-抗体的多重检测实验以及数据提取和分析方法。基于纸张的材料的使用使得该设备能够以具有成本效益的方式生产，并且盒式生产允许均匀的阵列形成。IgG 和 CRP 用于评估设备的性能，作为常见的生物标志物。本研究中提出的设备目前正在研究中。为了验证本研究中提出的 VFA 设备的多目标检测能力，使用了两种类型的抗原-抗体（人 IgG 和人 CRP）。在肉眼下，IgG 的检测限为 0.15μg/mL，CRP 的检测限为 0.19μg/mL。此外，通过 IgG 和 CRP 抗原证实了该设备结构不会引起交叉反应。总之，本研究中提出的 VFA 设备采用一步分析过程，并且已经证实它可以同时检测多个目标。
