Department of Applied Chemistry, Graduate School of Engineering, Osaka Prefecture University, 1-1 Gakuen-cho Naka Sakai, Osaka, 599-8531, Japan.
Sysmex Corporation, 1-5-1 Wakinohamakaigandori Chuo, Kobe, Hyogo, 651-0073, Japan.
Anal Sci. 2021 Oct 10;37(10):1473-1476. doi: 10.2116/analsci.21N011. Epub 2021 Apr 30.
In this paper, we report a single-step trypsin inhibitor assay on a microchannel array device immobilizing enzymes and substrates by inkjet printing. The microdevice is composed of a poly(dimethylsiloxane) (PDMS) microchannel array that immobilizes trypsin and fluorescent substrates as reactive reagents at the two bottom corners of a microchannel. Inkjet printers allow simple, accurate, and position-selective immobilization of reagents as nanoliter spots. Therefore, plural reactive reagents, such as enzymes and substrates, can be separately immobilized at different positions in the same microchannel without mixing, and thus allowing for single-step operation by simply introducing a sample solution through capillary action. Furthermore, reproducible fabrication and mass production of the device could be expected. In this study, the efficiency of an acidic solution as a spotting agent for protease immobilization to prevent decrease in the fluorescence intensity was confirmed. Additionally, single-step trypsin inhibitor screening was performed using three inhibitors. Finally, we investigated the storage stability of the device and confirmed that it remained stable for at least 10 days.
在本文中，我们报告了一种在微通道阵列设备上进行的单步胰蛋白酶抑制剂测定，该设备通过喷墨打印将酶和底物固定在微通道上。该微器件由聚二甲基硅氧烷（PDMS）微通道阵列组成，在微通道的两个底角固定胰蛋白酶和荧光底物作为反应试剂。喷墨打印机允许试剂以纳升级液滴的形式进行简单、准确和位置选择性的固定。因此，可以将多个反应试剂（如酶和底物）分别固定在同一微通道的不同位置，而不会混合，并且通过简单地通过毛细作用引入样品溶液即可实现单步操作。此外，可以预期该设备的可重复性制造和批量生产。在这项研究中，确认了酸性溶液作为蛋白酶固定点的效率，以防止荧光强度降低。此外，使用三种抑制剂进行了单步胰蛋白酶抑制剂筛选。最后，我们研究了该设备的存储稳定性，并证实其至少稳定 10 天。
