Toward a Practical Assay for Ion Channel Activity: Can a Silicon Device Support Planar Lipid Bilayers?

这项工作的目标是开发一种简单的可重复的方法来分析平面脂质双分子层中功能性离子通道的存在。改进脂质体离子通量测定和使用blm、贴片电极或全细胞的电测量将极大地促进生物物理和生化研究中离子通道活性的常规检测。如果单分子膜形成贴片夹持双分子膜的技术不是不可复制的，它将是很好的技术。我们建议测试一种形成稳定的平面双分子层的新方法，这种方法将很好地适用于这样一个新的分析系统——这是支持平面亲水性表面上的双分子层，这些双分子层由每个含有一个Ag/AgCl电极的小单孔穿孔。首先，我们将制造具有小亲水性孔的平面硅和玻璃结构。脂质双分子层将在这些孔洞上形成，方法是将装置两次穿过由带有或不带有离子通道的脂质单层覆盖的空气-水界面。烟碱乙酰胆碱受体在这种双分子层中的功能测量将用传统的电生理技术进行。

The goal of the work is development of a simple reproducible method for assaying for the presence of functional ion channels in planar lipid bilayers. Improving upon liposomal ion flux assays and electrical measurements employing BLMs, patch electrodes, or whole cells would greatly facilitate routine testing for ion channel activity in biophysical and biochemical research. The technique of patch clamping bilayers formed from monolayers would serve well if it were not so irreproducible. We propose to test a new approach at forming stable planar bilayers that will lend itself well to such a new assay system--this is to support bilayers on planar hydrophilic surfaces perforated by small single orifices containing one Ag/AgCl electrode each. First we will fabricate planar silicon and glass structures with small hydrophilic holes. Lipid bilayers will then be formed across those holes by passing the device twice through an air-water interface covered by a lipid monolayer with or without ion channels. Measurement of the function of the nicotinic acetylcholine receptor in such bilayers will be made with conventional electrophysiological techniques.