Molecular Biosensor Signal Amplification via Restriction Enzyme Cleavage
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Solution Overview
Problem
Current methods for detecting and measuring specific proteins in biological and environmental samples are complex, time-consuming, and lack efficient signal amplification, making them unsuitable for rapid and high-throughput analysis.
Innovation Solution
A molecular biosensor system comprising two or three components, including epitope-binding agent constructs and an oligonucleotide construct with restriction enzyme recognition sites, which amplifies signals through target-induced association and subsequent restriction enzyme cleavage, enabling the detection of proteins, carbohydrates, nucleic acids, and other analytes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional detection methods (gas chromatography, mass spectroscopy, DNA sequencing, immunoassays) are used to detect specific molecules, then detection accuracy can be achieved, but the process becomes complex, time-consuming, and expensive
Solution Approach 1:
The biosensor system is divided into separate functional modules: epitope-binding agents for target recognition, oligonucleotide constructs with restriction enzyme recognition sites for signal generation, and restriction enzymes for cleavage. This segmentation allows each component to perform its specific function efficiently, simplifying the overall detection process while maintaining high accuracy
Solution Approach 2:
The patent introduces oligonucleotide constructs as intermediary elements that bridge target recognition and signal detection. These constructs contain restriction enzyme recognition sites that serve as intermediaries between the epitope-binding agents and the final detectable signal, enabling a streamlined detection pathway that reduces complexity
2Measurement precision
If traditional immunoassays are used for protein detection, then sensitivity can be achieved, but the method is not well adapted for rapid, high throughput parallel detection
Solution Approach 1:
The biosensor system uses universal oligonucleotide constructs with restriction enzyme recognition sites that can be paired with various epitope-binding agents targeting different proteins. This multi-functional design allows the same core detection mechanism to be applied across multiple analytes, enabling high-throughput parallel detection while maintaining sensitivity
Solution Approach 2:
The patent employs restriction enzyme cleavage as a parameter change mechanism that produces discrete, measurable outcomes. This binary cleavage event (cleaved/not cleaved) creates a clear signal that can be rapidly detected and quantified, significantly increasing throughput compared to traditional continuous readout methods
3Measurement precision
If signal amplification is added to detection methods, then detection sensitivity improves, but the process becomes more complex and time-consuming
Solution Approach 1:
The oligonucleotide constructs are pre-designed with restriction enzyme recognition sites incorporated into their sequences. This preliminary arrangement of components allows the restriction enzyme to immediately cleave the oligonucleotide upon target binding, generating a signal amplification effect without requiring additional time-consuming steps
Solution Approach 2:
The system uses the target molecule itself to trigger the signal amplification process through spontaneous association with epitope-binding agents, which then directly activates restriction enzyme cleavage. This self-service mechanism eliminates the need for external amplification reagents or additional processing steps, maintaining rapid detection while achieving 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
The biosensor system provides a rapid, homogeneous means for detecting target molecules with enhanced signal amplification, facilitating sensitive and efficient analysis of various analytes, including proteins and nucleic acids, in biological and environmental samples.
Implementation Method 1
an oligonucleotide construct comprising a restriction enzyme recognition site
Data Source
AI summary
The present invention provides molecular biosensors capable of signal amplification, and methods of using the molecular biosensors to detect the presence of a target molecule.