Ion Channel Screening Using Cell Viability Readouts
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Solution Overview
Problem
Current methods for screening compounds targeting ion channels face limitations in measurement accuracy and throughput, with conventional fluorescent membrane potential measurement methods being inaccurate and automated patch clamp methods being inefficient and costly.
Innovation Solution
A screening system using cells with inhibited voltage-dependent Na ion channels and activated K channels, where cell death or viability indicates the action of test compounds on ion channels, allowing for high-throughput and accurate detection of compound interactions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional fluorescent membrane potential measurement methods are used, then large numbers of specimens can be evaluated, but measurement accuracy and applicable range are limited
Solution Approach 1:
The patent replaces the conventional fluorescent membrane potential measurement method with a cell death-based screening method. Instead of measuring membrane potential changes using fluorescent dyes, the invention uses cell viability/death as a readout to detect ion channel modulator activity, thereby achieving both high throughput and improved accuracy.
Solution Approach 2:
The patent changes the measurement parameter from membrane potential (which has limited accuracy and range) to cell viability (which provides more accurate and broader detection capability). This parameter change enables simultaneous achievement of high throughput and high accuracy in ion channel screening.
2Measurement precision
If patch clamp method is used, then high measurement accuracy and large amount of information can be obtained, but efficiency is poor due to small number of specimens measurable at one time
Solution Approach 1:
The patent divides the screening process into multiple parallel wells, each containing cells expressing the target ion channel. This segmentation allows simultaneous evaluation of multiple specimens in parallel, achieving high throughput while maintaining the accuracy of the patch clamp methodology through individual well analysis.
Solution Approach 2:
The patent creates multiple copies of the ion channel expression system in different wells, allowing parallel measurement of multiple specimens. This copying approach enables high throughput screening while preserving the high accuracy characteristics of the original patch clamp method.
3Productivity
If automated patch clamp method is used, then large number of specimens can be evaluated simultaneously, but efficiency of patch is low and equipment and running costs are high
Solution Approach 1:
The patent replaces expensive, complex automated patch clamp equipment with a simpler, more economical cell-based assay system. The methodology uses standard microplate formats and conventional detection methods, eliminating the need for costly automated patch clamp machinery while maintaining high throughput capability.
Solution Approach 2:
The patent substitutes the complex mechanical and electrical systems of automated patch clamp with a biochemical cell viability assay. This replacement eliminates expensive equipment requirements and reduces operational complexity while achieving simultaneous evaluation of large numbers of specimens.
4Productivity
If screening system targets ion channels with endogenous ligands being ions, then high-throughput is important, but ligand optimization is difficult resulting in need for measurement accuracy simultaneous to high throughput
Solution Approach 1:
The patent changes the detection parameter from membrane potential to cell viability, enabling simultaneous achievement of high throughput and high accuracy. This parameter change facilitates ligand optimization by providing reliable, quantitative readouts that work effectively with ion channel modulators, addressing the difficulty of optimizing ligands with ion channel targets.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach enables efficient and accurate detection of compound interactions with ion channels, overcoming the limitations of existing methods by using cell death or viability as an indicator, thus facilitating high-throughput screening with improved measurement accuracy.
Implementation Method 1
cells which act on a target ion channel, which retain at least one first DNA encoding a voltage-dependent Na ion channel that has been inhibited from being inactivated, and in which a K channel has been activated so that a resting membrane potential becomes deeper in a negative direction
Implementation Method 2
a K channel has been activated so that a resting membrane potential becomes deeper in a negative direction
Implementation Method 3
the cells which undergo cell death due to an ion concentration change in the cells, and typically due to an increase in intracellular sodium (Na) ion concentration
Data Source
AI summary
An object of the present invention is to provide a screening system that targets ion channels and has superior efficiency. The present invention provides a material for screening for compounds that act on a target ion channel, comprising cells which retain at least one first DNA encoding a voltage-dependent Na ion channel that has been inhibited from being inactivated, and in which a K ion channel has been activated so that a resting membrane potential becomes deeper in a negative direction.