Instrumentation Development for Parallel Measurement of an Ion Channel Array Chip

离子通道阵列芯片并行测量仪器的开发

• 批准号: 7999320
• 负责人: Jason L. Poulos
• 金额: $ 12.83万
• 依托单位: LIBREDE, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-07 至 2011-03-06
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7999320
• 关键词: Amplifiers; Biological Assay; Cell Culture Techniques; Cell Line; Cells; Collection; Detection; Development; Drug Formulations; Elements; Equipment; Goals; Housing; Human Resources; Injection of therapeutic agent; Ion Channel; Laboratories; Legal patent; Libraries; Lipid Bilayers; Liquid substance; Measurement; Measures; Membrane; Membrane Lipids; Methods; Molds; Monitor; Noise; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Physiological; Preclinical Drug Evaluation; Process; Proteins; Research; Resolution; Safety; Screening procedure; Shipping; Ships; Signal Transduction; Site; Small Business Innovation Research Grant; Solutions; System; Technology; Technology Transfer; Testing; Time; Training; Work; base; commercialization; cost; drug discovery; experience; high throughput screening; improved; instrument; instrumentation; patch clamp; product development; prototype; public health relevance; reconstitution; response; technology development; voltage

DESCRIPTION (provided by applicant): Electrophysiological assays of ion channels for pharmaceutical discovery and safety screening are problematic to perform in high throughput because the ion channels must be incorporated into a lipid bilayer membrane to enable measurement of their ionic conductance. As a result, there are currently no high quality, high throughput assays for ion channel screening. Recent developments of automated patch clamp instrumentation are still over two orders of magnitude lower throughput than conventional drug screening for soluble proteins and also require expensive instrumentation, specialized cell lines, and consumables. For existing methods of ion channel screening, there is a large gap in information quality, throughput, and cost. Librede Inc. is developing an alternative technology for ion channel measurement in which the ion channels are reconstituted in artificial lipid bilayer membranes. Librede's patent pending formulation of cell-free artificial bilayers has the potential for significantly higher throughput and lower consumable costs, while requiring less expensive equipment and trained personnel. Librede was founded by the UCLA inventors of this technology; we are now working to transfer this technology from the academic laboratory and develop it commercially. In preliminary work, we have developed inexpensive, disposable, shippable bilayer array chips capable of supporting ion channel measurement in 48 sites simultaneously. We have measured bilayers and ion channels in these chips previously using a multiplexed single channel amplifier; in the work proposed here, we aim to demonstrate higher throughput by measuring all sites in the chip simultaneously using a 48 channel patch clamp amplifier. Simultaneous measurement will be a key test of our technology; we will explore the effects of noise, crosstalk, and bilayer size on our ability to measure bilayers and incorporated ion channels over the entire chip at once. The noise and bandwidth achieved with our system using this instrument will determine our signal detection and temporal resolution and determine the feasibility of this platform for high throughput measurement of ion channels as well as paths to improvements in performance. The ability to measure ion channels in bilayer arrays in parallel will be a major milestone in the development of this technology toward commercialization, and is a preliminary step of the full demonstration of our final product. We will use the materials and experience gained in our Phase I research for Phase II development in which we will integrate low cost mass-producible injection molded shippable bilayer chips with automatable instrumentation for fluid handling and parallel ion channel measurement. This will bring us closer to our goal of reducing the cost and expertise required for ion channel screening by eliminating cell culture and cellular manipulation in pharmaceutical screening, thus significantly increasing throughput and decreasing costs, enabling more effective searches for ion channel drugs. PUBLIC HEALTH RELEVANCE: Measurement of ion channel interactions with drugs is a key process in drug discovery and drug safety screening, but due to the difficulty in working with ion channels the existing processes used are slow, laborious, and expensive. Librede has recently developed a platform for ion channel measurement which is much less expensive and much easier to use, based on lipid membranes that can be shipped-a world first. We propose here to develop instrumentation for simultaneously measuring arrays of ion channels contained in these membranes.

描述（由申请人提供）：用于药物发现和安全性筛选的离子通道的电生理学测定在高通量下进行是有问题的，因为离子通道必须掺入脂质双层膜中以能够测量其离子电导。因此，目前没有用于离子通道筛选的高质量、高通量测定。自动化膜片钳仪器的最新发展仍然比用于可溶性蛋白的常规药物筛选的通量低两个数量级以上，并且还需要昂贵的仪器、专门的细胞系和消耗品。对于现有的离子通道筛选方法，在信息质量、通量和成本方面存在很大差距。利布雷德公司正在开发一种用于离子通道测量的替代技术，其中离子通道在人工脂质双层膜中重构。Librede正在申请专利的无细胞人工双层制剂有可能显着提高通量并降低消耗成本，同时需要更便宜的设备和训练有素的人员。Librede由加州大学洛杉矶分校这项技术的发明者创立;我们现在正在努力从学术实验室转移这项技术并进行商业开发。在初步工作中，我们已经开发出廉价的，一次性的，可装运的双层阵列芯片，能够支持离子通道测量在48个网站同时进行。我们已经测量了双层和离子通道在这些芯片以前使用多路复用单通道放大器，在这里提出的工作中，我们的目标是通过测量所有网站的芯片同时使用48通道膜片钳放大器，以证明更高的吞吐量。同时测量将是我们的技术的一个关键测试;我们将探索噪声，串扰和双层大小对我们测量双层和整个芯片上的离子通道的能力的影响。使用该仪器的系统实现的噪声和带宽将确定我们的信号检测和时间分辨率，并确定该平台用于离子通道的高通量测量的可行性以及性能改进的途径。平行测量双层阵列中离子通道的能力将是该技术商业化发展的一个重要里程碑，也是我们最终产品全面展示的第一步。我们将使用在第一阶段研究中获得的材料和经验进行第二阶段开发，在第二阶段开发中，我们将整合低成本可大规模生产的注塑成型可运输双层芯片，以及用于流体处理和平行离子通道测量的自动化仪器。这将使我们更接近我们的目标，即通过消除药物筛选中的细胞培养和细胞操作来降低离子通道筛选所需的成本和专业知识，从而显着提高通量并降低成本，从而能够更有效地搜索离子通道药物。 公共卫生关系：离子通道与药物相互作用的测量是药物发现和药物安全性筛选中的关键过程，但是由于离子通道工作的困难，所使用的现有过程缓慢、费力且昂贵。Librede最近开发了一个离子通道测量平台，它更便宜，更容易使用，基于可以运输的脂质膜，这是世界上第一个。我们建议在这里开发仪器，同时测量这些膜中包含的离子通道阵列。