Kaminaga Maho, Ishida Tadashi, Kadonosono Tetsuya, Kizaka-Kondoh Shinae, Omata Toru
Department of Mechanical Engineering, School of Engineering, Tokyo Institute of Technology, Kanagawa 226-8503, Japan.
Department of Life Science and Technology, School of Life Science and Technology, Tokyo Institute of Technology, Kanagawa 226-8503, Japan.
Micromachines (Basel). 2019 Jan 9;10(1):41. doi: 10.3390/mi10010041.
This paper proposes a microfluidic device for screening molecules such as aptamers, antibodies, proteins, etc. for target cell-specific binding molecules. The discovery of cancer cell-specific binding molecules was the goal of this study. Its functions include filtering non-target cell-binding molecules, trapping molecules on the surface of target cells, washing away unbound molecules, and collecting target cell-specific binding molecules from target cells. These functions were effectively implemented by using our previously developed micro pillar arrays for cell homogeneous dispersion and pneumatic microvalves for tall microchannels. The device was also equipped with serially connected filter chambers in which non-target cells were cultured to reduce the molecules binding to non-target cells as much as possible. We evaluated the performance of the device using cancer cell lines (N87 cells as target cells and HeLa cells as non-target cells) and two fluorescent dye-labeled antibodies: Anti-human epidermal growth factor receptor 2 (anti-HER2) antibody that binds to target cells and anti-integrin antibody that binds to non-target cells. The results showed that the device could reduce anti-integrin antibodies to the detection limit of fluorescent measurement and collect anti-HER2 antibodies from the target cells.
本文提出了一种微流控装置,用于筛选适体、抗体、蛋白质等分子,以寻找靶细胞特异性结合分子。发现癌细胞特异性结合分子是本研究的目标。其功能包括过滤非靶细胞结合分子、捕获靶细胞表面的分子、洗去未结合的分子以及从靶细胞中收集靶细胞特异性结合分子。通过使用我们先前开发的用于细胞均匀分散的微柱阵列和用于高大微通道的气动微阀,有效地实现了这些功能。该装置还配备了串联连接的过滤室,在其中培养非靶细胞,以尽可能减少与非靶细胞结合的分子。我们使用癌细胞系(以N87细胞为靶细胞,HeLa细胞为非靶细胞)和两种荧光染料标记的抗体评估了该装置的性能:与靶细胞结合的抗人表皮生长因子受体2(抗HER2)抗体和与非靶细胞结合的抗整合素抗体。结果表明,该装置可以将抗整合素抗体降低到荧光测量的检测限,并从靶细胞中收集抗HER2抗体。