Droplet-based artificial membrane technology for high throughput cell-free ion ch

描述(申请人提供)：离子通道药物发现和安全筛选缓慢且昂贵，因此，目前还没有用于离子通道筛选的高质量、高通量的检测方法。Librede Inc.正在开发用于离子通道测量的替代无电池技术，这种技术具有提高吞吐量和降低成本的潜力。在前期工作中，我们已经展示了一种与自动运动控制硬件兼容的人造细胞膜的形成和测量平台。对这些膜的测量表明，它们能够容纳具有与科学文献相匹配的性质的离子通道。该平台的最初工作是在不断监测和反馈的情况下手动定位和组装部件。尽管这足以证明这项技术，但还不足以证明其适用于高产量和自动化过程。在这里，我们建议构建一种机械夹具，它将允许膜组件的“盲装”组装，模拟自动化过程。有了这个夹具，我们将使成膜设备循环运行，测量生成的人造膜的性能，以确定重复性程度，从而确定该工艺是否适合自动化和高通量。我们设计了一种与行业标准384井液处理系统兼容的板材，并对我们最初的成膜设备进行了改进，允许微小的机械定位误差。在拟议的工作中，我们将使用该牌照，并确定这些修改是否足够或需要进一步改进。我们将使仪器循环500多次，测量产生的膜的产量和它们重建离子通道的能力。开发的工艺将是可扩展的整个384孔板和机械夹具，易于与运动控制机器人一起使用，将我们的平台定位为第二阶段，使其适用于高吞吐量自动化。 公共卫生相关性：测量离子通道与药物的相互作用是药物筛选的关键过程，但目前的技术缓慢且昂贵。我们的团队最近开发了一种与机器人自动化兼容的人造细胞膜的形成方法。我们在这里建议开发工具和过程，使这项技术的缩放和循环，从而实现高通量离子通道测量。

DESCRIPTION (provided by applicant): Ion channels pharmaceutical discovery and safety screening is slow and expensive and as a result, there are currently no high quality, high throughput assays for ion channel screening. Librede Inc. is developing alternative cell-free technologies for ion channel measurement which have the potential for higher throughput and lower cost. In preliminary work, we have demonstrated a platform for formation and measurement of artificial cell membranes that is compatible with automated motion control hardware. Measurements of these membranes show that they are able to house ion channels with properties matching the scientific literature. The initial work with this platform utilized manual positioning and assembly of the components under constant monitoring and feedback. Although this was sufficient to demonstrate the technology, it is not sufficient to demonstrate its suitability for high throughput and automation processes. Here we propose to construct a mechanical jig which will allow assembly of the membrane components "blind", simulating automated processes. With this jig, we will cycle the membrane formation apparatus, measuring the properties of the resultant artificial membranes to determine the degree of reproducibility and therefore the suitability of this process for automation and high throughput. We have designed a plate which is compatible with industry standard 384 well fluid handling systems and contains a modification of our initial membrane formation apparatus which tolerates minor mechanical positioning errors. In the proposed work, we will use the plate and ascertain whether these modifications are sufficient or require further improvement. We will cycle the apparatus over 500 times, measuring the yield of the resulting membranes and their ability to reconstitute ion channels. The processes developed will be scalable an entire 384 well plate and the mechanical jig easily adapted for use with motion control robotics, positioning our platform for Phase II adapting it for high throughput automation. PUBLIC HEALTH RELEVANCE: Measurement of ion channel interactions with drugs is a key process in drug screening, but current technologies are slow and expensive. Our team has recently developed a method for formation of an artificial cell membrane that is compatible with robotic automation. We propose here to develop tools and processes that enable the scaling and cycling of this technology resulting in high throughput ion channel measurement.