Kawai Yuko, Shirai Akihiro, Kakuta Masaya, Idegami Kotaro, Sueyoshi Kenji, Endo Tatsuro, Hisamoto Hideaki
Department of Applied Chemistry, Graduate School of Engineering, Osaka Prefecture University, Osaka, Japan.
Sysmex Corporation, Hyogo, Japan.
Front Chem. 2020 Dec 21;8:612132. doi: 10.3389/fchem.2020.612132. eCollection 2020.
In this study, we report an inkjet printing-based method for the immobilization of different reactive analytical reagents on a single microchannel for a single-step and homogeneous solution-based competitive immunoassay. The immunoassay microdevice is composed of a poly(dimethylsiloxane) microchannel that is patterned using inkjet printing by two types of reactive reagents as dissolvable spots, namely, antibody-immobilized graphene oxide and a fluorescently labeled antigen. Since nanoliter-sized droplets of the reagents could be accurately and position-selectively spotted on the microchannel, different reactive reagents were simultaneously immobilized onto the same microchannel, which was difficult to achieve in previously reported capillary-based single-step bioassay devices. In the present study, the positions of the reagent spots and amount of reagent matrix were investigated to demonstrate the stable and reproducible immobilization and a uniform dissolution. Finally, a preliminary application to a single-step immunoassay of C-reactive protein was demonstrated as a proof of concept.
在本研究中，我们报告了一种基于喷墨打印的方法，用于将不同的反应性分析试剂固定在单个微通道上，以进行基于均相溶液的单步竞争性免疫测定。免疫测定微器件由聚二甲基硅氧烷微通道组成，该微通道通过喷墨打印将两种类型的反应性试剂作为可溶解斑点进行图案化，即固定有抗体的氧化石墨烯和荧光标记的抗原。由于试剂的纳升级液滴可以准确且位置选择性地滴印在微通道上，不同的反应性试剂同时固定在同一微通道上，这在先前报道的基于毛细管的单步生物测定装置中难以实现。在本研究中，研究了试剂斑点的位置和试剂基质的量，以证明稳定且可重复的固定以及均匀的溶解。最后，作为概念验证，展示了其在C反应蛋白单步免疫测定中的初步应用。
