SBIR-STTR Award

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.

Thesaurus Terms:
Amplifiers;Assay;Bioassay;Biologic Assays;Biological Assay;Cell Communication And Signaling;Cell Culture Techniques;Cell Line;Cell Lines, Strains;Cell Membrane Lipids;Cell Signaling;Cellline;Cells;Collection;Detection;Development;Drug Evaluation, Preclinical;Drug Formulations;Drug Screening;Drugs;Elements;Equipment;Evaluation Studies, Drug, Pre-Clinical;Evaluation Studies, Drug, Preclinical;Formulation;Formulations, Drug;Fungi, Filamentous;Goals;High Throughput Assay;Housing;Human Resources;Injection Of Therapeutic Agent;Injections;Instrumentation, Other;Intracellular Communication And Signaling;Ion Channel;Ionic Channels;Laboratories;Legal Patent;Libraries;Lipid Bilayers;Liquid Substance;Manpower;Measurement;Measures;Medication;Membrane;Membrane Channels;Membrane Lipids;Methods;Molds;Monitor;Noise;Patents;Performance;Pharmaceutic Preparations;Pharmaceutical Agent;Pharmaceutical Preparations;Pharmaceuticals;Pharmacologic Substance;Pharmacological Substance;Phase;Physiologic;Physiological;Preclinical Drug Evaluation;Process;Proteins;Research;Resolution;Sbir;Sbirs (R43/44);Safety;Screening Procedure;Shipping;Ships;Signal Transduction;Signal Transduction Systems;Signaling;Site;Small Business Innovation Research;Small Business Innovation Research Grant;Solutions;System;System, Loinc Axis 4;Technology;Technology Transfer;Testing;Time;Training;V (Voltage);Work;Base;Biological Signal Transduction;Commercialization;Cost;Cultured Cell Line;Drug Discovery;Drug/Agent;Experience;Fluid;Gene Product;High Throughput Screening;Improved;Instrument;Instrumentation;Lipid Bilayer Membrane;Liquid;Membrane Structure;Patch Clamp;Personnel;Product Development;Prototype;Public Health Relevance;Reconstitute;Reconstitution;Response;Screening;Screenings;Technology Development;Voltage

Ion channels are important drug targets-present in every cell, they play key roles in a wide range of physiological processes including the cardiac cycle and neural activity. As a result, unintended drug interactions with ion channels are also of critical importance, requiring the screening of all drug candidates against specific ion channels. Unfortunately, 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's goal is to develop new technologies that increase the efficiency of early stage pharmaceutical research and development. To this end, Librede 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 Phase I, we measured a 48 membrane array plate simultaneously with a multichannel amplifier and verified that the measurement performance was equal to or exceeded competitive automated patch clamp instruments. In the Phase II work proposed here, we will build on this work by integrating our Phase I instrumentation with fluid handling and motion control hardware to construct an automated workstation for cell-free ion channel screening using Librede's artificial membrane technology. We will consult with a pharmaceutical screening industry automation expert in the design, construction, and operation of our workstation and confirm that it can process and measure Librede's artificial membrane plates similarly to our previous work. To demonstrate its capabilities, we will perform a limited screen of CLIC1, an ion channel implicated in Alzheimer's disease. CLIC1 is particularly difficult to screen with conventional patch clamp, which makes this an ideal proof-of-concept application of our platform. We will work with the Schmidt group at UCLA, who have previously demonstrated the measurement of CLIC1 in artificial membranes. We will run the workstation in full automation mode, measuring ion channel activity as a function of drug concentration for a 72 compound ion channel-targeted drug library. This demonstration is essentially identical to an industry screen and is a major milestone for Librede's development of our artificial membrane technology, which promises an order of magnitude improvement in cost and throughput for ion channel screening.

Public Health Relevance:
Librede is developing a novel cell-free ion channel screening platform that has the potential to significantly reduce costs and increase throughput of drug discovery and safety screening for ion channels. We will develop an ion channel measurement workstation that is capable of completely automated processing and measurement of Librede's ion channel plates after the addition of drug compounds modulating the channel activity. We are validating this workstation by screening compounds for activity against the CLIC1 human ion channel which is thought to play an important role in Alzheimer's disease.