Aptazyme Sensor Device Inhibitory Component Shelf-Life
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Solution Overview
Problem
Aptazyme sensors face challenges with increased sensitivity leading to residual background activity, affecting shelf-life and commercial use, and unspecific binding to surfaces causing false positives and decreased sensitivity, along with limitations in parallelizing sensor reactions in a single vessel.
Innovation Solution
The aptazyme sensor devices incorporate an aptamer component, a ribozyme component with switchable enzymatic activity, a communication component, and an inhibitory or signalling component with a metal nanoparticle, allowing selective inhibition and activation of enzymatic activity through plasmon resonance excitation, and post-synthetic chemical modification to reduce unspecific binding.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If aptazyme sensors are designed with high sensitivity, then detection capability is improved, but residual background activity increases which reduces shelf-life
Solution Approach 1:
The patent applies preliminary anti-action by introducing an inhibitory component that preemptively blocks the ribozyme's substrate binding site before detection occurs. This inhibitory component binds to the ribozyme in advance, preventing any residual catalytic activity during storage. When detection is needed, the inhibitory component is removed, activating the ribozyme only at that moment. This approach allows high sensitivity detection capability while eliminating background activity that would otherwise reduce shelf-life.
Solution Approach 2:
The patent implements preliminary action by pre-assembling the complete sensor device including aptamer, ribozyme, and communication module in an inactive state. The inhibitory component is introduced beforehand to suppress any premature enzymatic activity. This preliminary preparation ensures the sensor maintains high detection sensitivity while remaining stable during storage, as the inhibitory component prevents unwanted catalysis until the moment of use.
2Measurement precision
If aptamer components are increased in sensitivity, then ligand detection is improved, but unspecific binding to surfaces increases causing false positives
Solution Approach 1:
The patent applies the taking out principle by separating the aptamer from potential sources of unspecific binding (surfaces). Instead of allowing the aptamer to bind directly to surfaces, the invention uses a communication module that translates conformational changes in the aptamer into ribozyme activation. This extraction of the aptamer from direct surface contact eliminates unspecific binding while preserving specific ligand detection capability.
Solution Approach 2:
The patent introduces an intermediary system consisting of the communication module and ribozyme. The aptamer binds specifically to its target ligand, and this binding event is transmitted through the communication module to activate the ribozyme. This intermediary chain ensures that only specific aptamer-ligand interactions lead to signal generation, while unspecific surface binding of the aptamer does not trigger false positives, as the communication module only responds to the specific conformational change induced by target ligand binding.
3Productivity
If multiple sensor reactions are parallelized in a single vessel, then productivity is improved, but cross-interference between reactions increases
Solution Approach 1:
The patent applies segmentation by dividing the detection system into independent modular units, each consisting of an aptamer-specific to a particular ligand, a communication module, and a ribozyme. Each module operates independently within the same vessel, eliminating cross-interference between different ligand detection reactions. This modular segmentation enables parallel detection of multiple ligands while maintaining the reliability and accuracy of each individual detection channel.
4Duration of action of stationary object
If inhibitory component is added to reduce background activity, then shelf-life is improved, but device complexity increases
Solution Approach 1:
The patent applies merging by combining the inhibitory component with other sensor elements into an integrated assembly. The inhibitory component is designed to work seamlessly with the ribozyme and communication module as a unified system. This merging approach minimizes the increase in device complexity, as the inhibitory component is not merely an additive element but an integrated part of the sensor's operational mechanism, working in concert with the other components to achieve both extended shelf-life and high detection sensitivity.
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 solution enhances shelf-life by minimizing residual activity, enables parallel detection of multiple ligands in a single reaction vessel, and reduces unspecific binding, improving sensor sensitivity and specificity.
Implementation Method 1
binding of the single nucleotide strand to the substrate binding site of component (b) is selectively interrupted after exciting plasmon resonance of the metal nanoparticle by irradiation of said metal nanoparticle with light
Data Source
AI summary
The present invention relates to aptazyme sensor devices comprising, in addition to the aptamer, ribozyme and communication components, a competitive inhibitory component with a metal nanoparticle or a competitive inhibitory and signalling component with a metal nanoparticle and a label, such that the enzymatic activity of the ribozyme is inhibited as long as the inhibitory or the inhibitory and signalling component is bound to the substrate binding site of the ribozyme. The aptazyme sensor devices of the invention have increased shelf-life and are suitable for the parallel detection of different ligands by using an array of aptazyme sensor devices utilizing inhibitory and inhibitory and signalling components with different metal nanoparticles. The present invention furthermore relates to the post-synthetic chemical modification of the aptamer component for avoiding unspecific binding.