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

用于高通量无细胞离子通道的基于液滴的人工膜技术

• 批准号: 8118324
• 负责人: Jason L. Poulos
• 金额: $ 16.38万
• 依托单位: LIBREDE, INC.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-15 至 2012-03-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8118324
• 关键词: Adopted; Artificial Membranes; Automation; Biotechnology; Cardiac; Cell membrane; Cells; Development; Drug Delivery Systems; Drug Interactions; Ensure; Equilibrium; Feedback; Freedom; Future; Hour; Housing; Industry; Ion Channel; Ions; Lateral; Licensing; Liquid substance; Literature; Maintenance; Manuals; Measurement; Measures; Mechanics; Membrane; Methods; Minor; Modification; Monitor; Motion; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Physiological Processes; Plastics; Play; Positioning Attribute; Potassium Channel; Preclinical Drug Evaluation; Process; Production; Property; Proteins; Reproducibility; Research; Risk; Robotics; Safety; Screening procedure; Simulate; System; Technology; Time; Transmembrane Transport; Work; base; blind; cost; design; drug candidate; drug discovery; health care quality; high throughput screening; high throughput technology; improved; instrumentation; membrane assembly; operation; patch clamp; reconstitution; relating to nervous system; tool

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.

描述（由申请人提供）：离子通道药物发现和安全筛查缓慢而昂贵，因此，目前尚无高质量，高吞吐量的离子通道筛选。 Librede Inc.正在开发用于离子通道测量的替代无细胞技术，该技术具有更高的吞吐量和较低成本的潜力。在初步工作中，我们展示了一个与自动运动控制硬件兼容的人造细胞膜形成和测量的平台。这些膜的测量表明，它们能够将离子通道带有与科学文献相匹配的特性。该平台的最初工作利用了在持续监视和反馈下的手动定位和组件的组件。尽管这足以证明这项技术，但不足以证明其适合高通量和自动化过程。在这里，我们建议构建一个机械夹具，该夹具将允许组装“盲”的膜组件，从而模拟自动化过程。使用此夹具，我们将循环膜形成设备，测量所得人工膜的性质，以确定可重复性的程度，从而确定该过程对于自动化和高吞吐量的适用性。我们设计了一个与行业标准384井流体处理系统兼容的板，并包含对我们初始膜形成设备的修改，该设备可容忍较小的机械定位误差。在拟议的工作中，我们将使用板并确定这些修改是否足够或需要进一步改进。我们将循环超过500次，以测量所得膜的产量及其重新构建离子通道的能力。开发的过程将是整个384井板的可扩展性，并且机械夹具很容易适用于运动控制机器人技术，从而将我们的II期平台定位为将其适应高吞吐量自动化。 公共卫生相关性：与药物的离子通道相互作用的测量是药物筛查的关键过程，但是当前的技术缓慢而昂贵。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方法。我们在这里建议开发工具和过程，以使该技术的缩放和循环能够进行高吞吐量离子通道测量。