Cell-Free NCX Sensor Chip for Rapid Solution Exchange
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Solution Overview
Problem
Current methods for determining the activity of Na+/Ca2+ exchanger (NCX) proteins are limited by the need for complex biological backgrounds that interfere with assays, and rapid solution exchange is not possible, leading to inaccurate measurements.
Innovation Solution
A cell-free electrophysiological assay using a sensor chip with immobilized NCX protein allows for rapid solution exchange and measurement of electrical currents, enabling efficient determination of NCX activity and modulation by compounds.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If patch-clamp experiments are used to monitor transporter-related currents, then measurement precision is improved, but device complexity and experimental difficulty increase due to requirement for living cells and complex biological backgrounds
Solution Approach 1:
The patent extracts the NCX protein from complex biological membranes and immobilizes it on a sensor chip surface, separating the protein of interest from interfering biological components. This allows measurement of transporter activity without the complexity of living cell systems while maintaining measurement precision through direct electrical current detection.
Solution Approach 2:
The patent introduces an intermediary sensor chip system that mediates between the NCX protein and the measurement device. The sensor chip provides a controlled environment with electrical electrodes that can detect currents generated by NCX activity, serving as an intermediary that simplifies the measurement system while maintaining precision.
2Measurement precision
If black lipid membrane systems are used, then measurement precision is improved, but productivity deteriorates due to inability to conduct rapid buffer exchange experiments
Solution Approach 1:
The patent implements a dynamic solution exchange system where buffer reservoirs can be rapidly changed to introduce different substrates or inhibitors. The system allows switching between activating and non-activating solutions, enabling kinetic studies and drug screening. This dynamic capability maintains measurement precision while dramatically improving productivity through rapid experiment iteration.
Solution Approach 2:
The patent changes the operational parameters of the system by implementing rapid buffer exchange capability. The system can switch between different solution conditions (activating vs non-activating buffers) to control NCX activity, enabling high-throughput screening and kinetic analysis that were not possible with static black lipid membrane systems.
3Productivity
If cell-free electrophysiological sensor chip is used, then productivity is improved through rapid solution exchange, but device complexity increases due to requirement for sensor chip manufacturing and protein immobilization
Solution Approach 1:
The patent applies preliminary action by pre-immobilizing the NCX protein on the sensor chip surface before experimentation. The protein is anchored to the chip in advance, creating a ready-to-use system that eliminates the need for complex in-situ protein insertion during experiments. This preliminary preparation simplifies the overall device while enabling rapid solution exchange and high-throughput screening.
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 provides accurate and efficient measurement of NCX protein activity and modulation, overcoming the limitations of complex biological backgrounds and enabling rapid solution exchange.
Implementation Method 1
a cell free electrophysiological sensor chip... detects the resulting transporter-related currents
Implementation Method 2
Most membrane transporters shift electrical charges while going through their transport cycle... These transporters are responsible for the generation and maintenance of ion gradients
Data Source
AI summary
The invention refers to a cell free assay for determining the activity of a Na+/Ca2+ exchanger (NCX) protein by means of a cell free electrophysiological sensor chip, a kit of parts comprising the sensor chip with a NCX protein as well as the manufacturing and use of the kit of parts.