Development of a disposable shippable chip for high throughput ion channel screen

开发用于高通量离子通道筛选的一次性可运输芯片

• 批准号: 7745583
• 负责人: Jason L. Poulos
• 金额: $ 13.05万
• 依托单位: LIBREDE, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7745583
• 关键词: Automation; Cells; Computer Interface; Contracts; Custom; Deposition; Development; Drug Delivery Systems; Drug Interactions; Electronics; Elements; Freezing; Goals; Industry; Injection of therapeutic agent; Ion Channel; Laboratories; Lipid Bilayers; Lipids; Liquid substance; Manufacturer Name; Measurable; Measurement; Membrane; Membrane Lipids; Methods; Molds; Pharmaceutical Preparations; Phase; Plastics; Preclinical Drug Evaluation; Process; Proteins; Research; Robotics; Safety; Screening procedure; Shipping; Ships; Small Business Innovation Research Grant; Solutions; Solvents; System; Technology; Temperature; Time; Transmembrane Transport; Work; base; cost; design; design and construction; drug candidate; drug discovery; experience; high throughput screening; improved; instrumentation; large scale production; patch clamp; prototype; public health relevance; reconstitution; success

DESCRIPTION (provided by applicant): Ion channels are important drug targets and unintended drug interactions with ion channels are also of critical importance, requiring the screening of all drug candidates. Conventional high throughput screening processes for soluble proteins are problematic to apply to ion channels because the channels must be incorporated into a lipid bilayer membrane and transport an ionic current to display their functionality. There are currently no high quality, high throughput assays for ion channel drug screening. Recent efforts to increase throughput have resulted in automated patch clamp systems but these are still over two orders of magnitude lower throughput than conventional drug screening technologies, requiring costly instrumentation, cells, and consumables. For existing methods of ion channel screening, there is a gap in information quality, throughput, and cost. An alternative method of ion channel measurement involves reconstituting them in artificial lipid bilayer membranes. In recent research at UCLA led by the PI, a new high-freezing point lipid membrane composition was developed enabling it to be frozen. When frozen, we showed that it was sufficiently robust to withstand shipping, a major breakthrough. These membranes, when packaged in inexpensive chips, have the potential to significantly change the way ion channel screening is done. Our company, Librede Inc., was formed by the UCLA team to further develop and explore the commercial potential of this technology, the first steps of which are proposed here in this Phase I SBIR proposal. Our ultimate goal is to create an inexpensive, disposable chip containing arrays of lipid membranes to enable low cost high throughput screening of ion channels. In the preliminary work at UCLA, the technology was demonstrated with the shipping of small numbers of membranes, with a net yield of 30%. In the proposed work, we will design and fabricate large array chips containing 48 membranes compatible with industry standard 96 well fluid handling robotics and pipetters. With these chips, we aim to demonstrate the viability of the membrane technology over a much larger scale producing over 3000 membranes. To increase their commercial viability, we will also seek to increase the yield by systematically changing the lipid concentration, solvent composition, solution volume, and thawing temperature. Although this space is potentially very large, the membrane arrays will electrically probed in an automated fashion using a custom-built electrical interface and multiplexer. We will use the materials and designs from Phase I to contract with a plastics manufacturer in Phase II to create an inexpensive, injection molded prototype. At that point, we will have demonstrated a chip able to perform industry standard ion channel screens for a significantly lower cost. These membrane array chips have the potential to increase throughput by several orders of magnitude and similarly decrease cost by several orders of magnitude-as a result, transforming the process of ion channel measurement and screening. PUBLIC HEALTH RELEVANCE: Measurement of ion channel interactions with drugs is a key process in drug discovery and drug safety screening. Due to the difficulty in working with ion channels, the existing processes used are slow, laborious, and expensive. Our team 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 inexpensive membrane array chips and use them for large scale production, demonstrating the feasibility of this technology for ion channel screening.

描述（由申请人提供）：离子通道是重要的药物靶标，而意外的药物与离子通道的相互作用也至关重要，需要筛查所有候选药物。可溶性蛋白的常规高通量筛选过程适用于离子通道是有问题的，因为必须将通道掺入脂质双层膜中并传输离子电流以显示其功能。目前，没有用于离子通道药物筛查的高质量，高通量测定。最新增加吞吐量的努力导致了自动化的斑块夹系统，但与传统的药物筛查技术相比，这些填充物的吞吐量仍然低两个数量级，需要昂贵的仪器，细胞和消耗品。对于现有的离子通道筛选方法，信息质量，吞吐量和成本存在差距。离子通道测量方法的另一种方法涉及将它们重建在人工脂质双层膜中。在PI领导的UCLA的最新研究中，开发了一种新的高冰点脂质膜组成，使其能够冷冻。冻结后，我们表明它足以承受运输，这是一个重大突破。这些膜在廉价的芯片中包装时，有可能显着改变离子通道筛选的方式。 UCLA团队成立了我们的公司Librede Inc.，以进一步开发和探索该技术的商业潜力，其第一步是在此阶段I SBIR提案中提出的。我们的最终目标是创建一个廉价的一次性芯片，其中含有脂质膜的阵列，以实现低成本的高成本筛选离子通道。在加州大学洛杉矶分校的初步工作中，该技术通过少量膜的运输进行了证明，净产量为30％。在拟议的工作中，我们将设计和制造大型阵列芯片，其中包含48种与行业标准96井的机器人和移液器兼容的膜。借助这些芯片，我们旨在证明膜技术在更大范围内产生3000多个膜的生存能力。为了提高其商业生存能力，我们还将通过系统地改变脂质浓度，溶剂组成，溶液体积和解冻温度来提高产量。尽管该空间可能非常大，但膜阵列将使用定制的电界面和多路复用器以自动方式进行电探测。我们将使用I阶段的材料和设计与II期塑料制造商收缩，以创建廉价的注射模制原型。到那时，我们将展示一个能够执行行业标准离子频道屏幕的芯片，成本明显降低。这些膜阵列芯片有可能增加多个数量级的吞吐量，并类似地将成本降低了几个数量级，从而改变了离子通道测量和筛选的过程。公共卫生相关性：离子通道与药物相互作用的测量是药物发现和药物安全筛查中的关键过程。由于难以使用离子通道，因此使用的现有过程缓慢，费力且昂贵。我们的团队最近开发了一个用于离子频道测量的平台，该平台基于可以发货的脂质膜，它首先是世界。我们在这里建议开发廉价的膜阵列芯片并将其用于大规模生产，以证明该技术对离子通道筛选的可行性。