Development of a disposable cell-free ion channel measurement platform for hERG s

描述(由申请人提供)：高通量地进行药物发现和安全筛选的离子通道分析是有问题的，因为离子通道必须结合到细胞膜中以允许测量它们的离子传输以确定它们的功能。因此，目前还没有用于离子通道筛选的高质量、高通量的分析方法。自动化膜片钳仪器的最新发展仍然比传统的可溶性蛋白质药物筛选的吞吐量低一个数量级以上，而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、吞吐量和成本方面存在很大差距。Librede Inc.正在开发一种使用人造细胞膜进行离子通道筛选的替代无细胞技术。Librede正在申请专利的无细胞人造膜配方可以实现更高的产量和更低的消耗品成本，同时需要更便宜的设备和训练有素的人员。Librede是由加州大学洛杉矶分校的研究人员和这项技术的发明者创立的。在这里提出的工作中，我们的目标是开发材料和方法来测量和包装我们的人工膜平台中的HERG通道。HERG是筛选的一个重要离子通道，因为药物在HERG通道上的非靶点活性可能导致心律失常和死亡；因此，所有候选药物，无论是什么靶点，都必须筛选以防止与HERG的相互作用。在这项工作中，我们将与加州大学洛杉矶分校的施密特小组合作，他们在初步工作中首次展示了人工膜中HERG的测量。我们的目标是确定HERG与Librede人工膜平台的兼容性，并确定优化HERG与我们的膜阵列板包装的长期储存的条件。最后，我们将通过在48膜阵列板上同时筛选HERG与已知的通道阻滞剂来演示我们的技术。这项工作是我们朝着降低离子通道筛选所需的成本和专业知识的目标迈出的关键一步。 与公共健康相关：我们最近开发了一种新的离子通道测量平台，该平台可以无限期存储、商业运输，并且只需要非常少的最终用户输入即可操作。我们的目标是用人类心脏离子通道HERG来验证该平台，以演示该技术在离子通道筛选中的应用。

DESCRIPTION (provided by applicant): Assaying ion channels for pharmaceutical discovery and safety screening is problematic to perform in high throughput because the ion channels must be incorporated into a cell membrane to allow measurement of their ionic transport to determine their functionality. 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 an order 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 cell-free technology for ion channel screening using artificial cell membranes. Librede's patent pending formulation of cell-free artificial membranes can enable higher throughput and lower consumable costs while requiring less expensive equipment and trained personnel. Librede was founded by UCLA researchers and the inventors of this technology. In the work proposed here, we aim to develop materials and methods for measurement and packaging of the hERG channel in our artificial membrane platform. hERG is an important ion channel to screen, since off-target activity of pharmaceuticals on the hERG channel can lead to cardiac arrhythmia and death; thus all drug candidates, irrespective of target, must be screened against interaction with hERG. For this work, we will collaborate with the Schmidt group at UCLA, who has demonstrated the measurement of hERG in artificial membranes in preliminary work, a first. We aim to determine the compatibility of hERG with Librede's artificial membrane platform and determine the conditions with optimize the long term storage of hERG packaged with our membrane array plates. Finally, we will perform a demonstration of our technology by screening hERG against known channel blockers over a 48 membrane array plate simultaneously. This work is a key step toward our goal of reducing the cost and expertise required for ion channel screening. PUBLIC HEALTH RELEVANCE: We have recently developed a novel ion channel measurement platform that may be indefinitely stored, commercially shipped, and requires very little input from the end user to operate. We aim to validate this platform with the human cardiac ion channel hERG to demonstrate the utility of this technology for ion channel screening.