Suzuki Yuma, Morioka Kazuhiro, Ohata Soichiro, Shimizu Tetsuhide, Nakajima Hizuru, Uchiyama Katsumi, Yang Ming
Graduate School of System Design, Tokyo Metropolitan University, 6-6 Asahigaoka, Hino, Tokyo 191-0065, Japan.
Department of Biomedical Analysis, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392, Japan.
Sensors (Basel). 2017 Jul 11;17(7):1608. doi: 10.3390/s17071608.
A film-stack reaction field with a micropillar array using a motor stirrer was developed for the high sensitivity and rapid enzyme-linked immunosorbent assay (ELISA) reaction. The effects of the incubation time of a protein (30 s, 5 min, and 10 min) on the fluorescence intensity in ELISAs were investigated using a reaction field with different micropillar array dimensions (5-µm, 10-µm and 50-µm gaps between the micropillars). The difference in fluorescence intensity between the well with the reaction field of 50-µm gap for the incubation time of 30 s and the well without the reaction field with for incubation time of 10 min was 6%. The trend of the fluorescence intensity in the gap between the micro pillars in the film-stack reaction field was different between the short incubation time and the long incubation time. The theoretical analysis of the physical parameters related with the biomolecule transport indicated that the reaction efficiency defined in this study was the dominant factor determining the fluorescence intensity for the short incubation time, whereas the volumetric rate of the circulating flow through the space between films and the specific surface area were the dominant factors for the long incubation time.
为实现高灵敏度和快速的酶联免疫吸附测定（ELISA）反应，开发了一种使用电机搅拌器的带有微柱阵列的膜堆反应场。使用具有不同微柱阵列尺寸（微柱之间的间隙为5 µm、10 µm和50 µm）的反应场，研究了蛋白质孵育时间（30秒、5分钟和10分钟）对ELISA中荧光强度的影响。对于30秒孵育时间的50 µm间隙反应场的孔与对于10分钟孵育时间的无反应场的孔之间的荧光强度差异为6%。在短孵育时间和长孵育时间之间，膜堆反应场中微柱之间间隙处的荧光强度趋势不同。对与生物分子传输相关的物理参数的理论分析表明，本研究中定义的反应效率是短孵育时间决定荧光强度的主要因素，而通过膜间空间的循环流动的体积速率和比表面积是长孵育时间的主要因素。
