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性筛选缓慢且昂贵，因此，目前没有用于离子通道筛选的高质量、高通量测定。利布雷德公司正在开发用于离子通道测量的替代无细胞技术，该技术具有更高通量和更低成本的潜力。在初步工作中，我们已经展示了一个与自动化运动控制硬件兼容的人工细胞膜形成和测量平台。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特性的离子通道。该平台的初始工作是在持续的监控和反馈下手动定位和组装组件。虽然这足以证明该技术，但不足以证明其适用于高通量和自动化工艺。在这里，我们建议构建一个机械夹具，这将允许组装的膜组件“盲”，模拟自动化过程。使用该夹具，我们将循环膜形成装置，测量所得人工膜的性质，以确定再现性的程度，从而确定该过程的自动化和高通量的适用性。我们已经设计了一种板，其与工业标准384井流体处理系统兼容，并且包含对我们的初始膜形成装置的修改，其容许微小的机械定位误差。在拟议的工作中，我们将使用该板，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次以上，测量所得膜的产量及其重建离子通道的能力。开发的工艺将可扩展到整个384孔板，机械夹具易于与运动控制机器人一起使用，将我们的平台定位于第二阶段，使其适应高通量自动化。 公共卫生关系：离子通道与药物相互作用的测量是药物筛选中的关键过程，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工细胞膜的形成方法。我们在这里建议开发工具和过程，使这种技术的缩放和循环，从而导致高通量离子通道测量。