The Graduate School for the Creation of New Photonics Industries, 1955-1 Kurematsu Nishi-ku, Hamamatsu, Shizuoka, 431-1202, Japan.
SYSTEC Corporation, 1-9-9 shinmiyakoda Kita-ku, Hamamatsu, Shizuoka, 431-2103, Japan.
Sci Rep. 2021 Sep 9;11(1):17934. doi: 10.1038/s41598-021-97237-z.
Artificial lipid bilayer single-channel recording technique has been employed to determine the biophysical and pharmacological properties of various ion channels. However, its measurement efficiency is very low, as it requires two time-consuming processes: preparation of lipid bilayer membranes and incorporation of ion channels into the membranes. In order to address these problems, we previously developed a technique based on hydrophilically modified gold probes on which are immobilized ion channels that can be promptly incorporated into the bilayer membrane at the same time as the membrane is formed on the probes' hydrophilic area. Here, we improved further this technique by optimizing the gold probe and developed an automated channel current measurement system. We found that use of probes with rounded tips enhanced the efficiency of channel current measurements, and introducing a hydrophobic area on the probe surface, beside the hydrophilic one, further increased measurement efficiency by boosting membrane stability. Moreover, we developed an automated measurement system using the optimized probes; it enabled us to automatically measure channel currents and analyze the effects of a blocker on channel activity. Our study will contribute to the development of high-throughput devices to identify drug candidates affecting ion channel activity.
人工脂质双层单通道记录技术已被用于确定各种离子通道的生物物理和药理学特性。然而,它的测量效率非常低,因为它需要两个耗时的过程:脂质双层膜的制备和离子通道掺入到膜中。为了解决这些问题,我们之前开发了一种基于亲水改性金探针的技术,在该技术中,离子通道固定在探针上,可以在探针亲水区域形成膜的同时迅速掺入到双层膜中。在这里,我们通过优化金探针进一步改进了这项技术,并开发了一种自动化的通道电流测量系统。我们发现,使用圆头探针可以提高通道电流测量的效率,并且在探针表面的亲水区域旁边引入疏水区可以通过提高膜的稳定性进一步提高测量效率。此外,我们使用优化的探针开发了一种自动化测量系统;它使我们能够自动测量通道电流并分析阻断剂对通道活性的影响。我们的研究将有助于开发高通量设备来鉴定影响离子通道活性的候选药物。