Center for BioMicrosystems, Korea Institute of Science and Technology, 5, Hwarang-ro 14-gil, Seongbuk-gu, Seoul, 02792, South Korea.
Department of Chemistry, Kookmin University, 77 Jeongneung-ro, Seongbuk-gu, Seoul, 02707, South Korea.
Biosens Bioelectron. 2019 Sep 15;141:111404. doi: 10.1016/j.bios.2019.111404. Epub 2019 Jun 6.
The bio-sensory organs of living creatures have evolved to have the best sensing performance. They have 3-dimensional protrusions that have large surface areas to accommodate a large number of membrane proteins such as ion channels and G-protein coupled receptors, resulting in high sensitivity and specificity to target molecules. From the perspective of mimicking this system, BLM, which has been used extensively as a platform for a single nanopore-based sensing systems, has some limitations, i.e., some residual solvent, low mechanical stability, small surface area for appropriate stability, and difficulty in high-throughput fabrication. Herein, to eliminate these limitations, a solvent-free, size-controllable, 3-dimensional free-standing lipid bilayer (3DFLB) structure array with high stability (∼130 h) and high density (∼300,000 cm) is proposed, and its structural advantages for efficient and rapid protein reconstitution, compared to BLM, is demonstrated by human 5-HT receptor assay as well as α-hemolysin assay. A continuous process of 3DFLB array fabrication, 5-HT reconstitution, and 5-HT detections in a microfluidic channel proves the applicability of the proposed structures as a highly-sensitive sensing platform mimicking bio-sensory organs.
生物活体的生物感应器官已经进化到具有最佳的感应性能。它们具有三维突起，具有较大的表面积，可以容纳大量的膜蛋白，如离子通道和 G 蛋白偶联受体，从而对目标分子具有高灵敏度和特异性。从模拟该系统的角度来看，BLM 已被广泛用作基于单个纳米孔的传感系统的平台，但它存在一些局限性，例如一些残留溶剂、低机械稳定性、适当稳定性的小表面积以及难以实现高通量制造。在此，为了消除这些限制，提出了一种无溶剂、尺寸可控的、具有高稳定性（约 130 小时）和高密度（约 300,000/cm）的三维独立脂质双层（3DFLB）结构阵列，并通过人 5-HT 受体测定和α-溶血素测定证明了其与 BLM 相比在高效快速蛋白质重组方面的结构优势。在微流道中进行 3DFLB 阵列制造、5-HT 重组和 5-HT 检测的连续过程证明了所提出的结构作为模拟生物感应器官的高灵敏度传感平台的适用性。
