Urea Biosensor Protein Binding Signal Transduction
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing urea detection methods are sensitive to inhibition and require multiple reagents or complex detectors, making them unreliable and cumbersome for practical applications.
Innovation Solution
Development of biosensors comprising a urea-binding protein with a reporter group that produces a detectable signal upon urea binding, allowing for rapid, reliable, and accurate urea detection without additional substrates or complex measurements, and maintaining functionality even after immobilization, desiccation, and extended storage.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If enzyme-based assays with urease are used to detect urea, then urea concentration can be measured, but the assay becomes sensitive to inhibition and requires multiple reagents or complex detectors
Solution Approach 1:
The patent extracts the urea detection function from complex enzyme-based systems and implements it through a simplified protein-binding approach. The urea-binding protein directly interacts with urea to produce a detectable signal, eliminating the need for urease enzymes and multiple reagents, thus reducing device complexity while maintaining reliability
Solution Approach 2:
The biosensor system is designed to be self-sufficient, where the urea-binding protein performs both the detection and signal generation functions. The system does not require external substrates or complex measurement mechanisms, allowing it to operate autonomously and simplifying the overall device architecture
2Measurement precision
If multiple reagents and complex detectors are used in urea detection, then measurement capability is achieved, but the system becomes cumbersome for practical applications
Solution Approach 1:
The patent segments the detection system into a single functional unit - the urea-binding protein that performs binding and signal generation in one component. This segmentation eliminates the need for multiple separate reagents and complex detectors, making the system easier to operate while maintaining measurement precision
Solution Approach 2:
The invention changes the fundamental detection parameter from enzymatic reaction products (requiring multiple reagents) to direct protein-ligand binding events. This parameter change simplifies the operational procedure while preserving accurate urea concentration measurement capability
3Productivity
If conventional biosensors are used, then urea detection is possible, but they require additional substrates or measurement of substrate consumption rates
Solution Approach 1:
The urea-binding protein serves itself by directly converting urea binding into a detectable signal without requiring external substrates. The protein's conformational change upon urea binding is directly measured, eliminating the need to monitor substrate consumption or product generation rates, thus simplifying the monitoring mechanism while maintaining high detection speed
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
The biosensors provide precise, reliable, and instantaneous urea detection in various samples, including complex fluids, with reduced costs and simplified handling, enabling widespread applicability in clinical, industrial, and environmental settings.
Implementation Method 1
The ligand comprises urea: and the ligand-binding protein includes a domain or region(s) of the protein that binds the urea. The domain or region involved in ligand binding is comprised of a plurality of residues, e.g., non-contiguous amino acids of the ligand-binding protein, which are contact points or sites of contact between the ligand and its cognate ligand-binding protein.
Implementation Method 2
The binding of urea to the urea-binding domain of the urea-binding protein causes a change in signaling by the reporter group. A reporter group that transduces a detectable signal may be attached to the urea-binding proteins (biosensors) described herein.
Implementation Method 3
In embodiments, detectable signal comprises a fluorescent, electrochemical, nuclear magnetic resonance (NMR), or electron paramagnetic resonance (EPR) signal.
Data Source
Figure 1A~1D
Figure 2A~2B
Figure 3
AI summary
The present subject matter provides urea biosensors as well as compositions, devices, and methods comprising such biosensors.