SPR Sensor NAzyme Activity Detection
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current enzyme-linked immunosorbent assay (ELISA) and high-resolution melting analysis (HRM) methods for protein and DNA detection are limited by extensive sample handling, need for trained staff, batch-to-batch variations, specific adsorption issues, and complexity, making them costly and less adaptable for diverse applications.
Innovation Solution
A surface plasmon resonance (SPR) sensor system that measures nucleic acid enzyme (NAzyme) activity using metallic nanoparticle-labelled DNA sequences, allowing for real-time monitoring of NAzyme cleavage activity and adaptation to various targets through a universal substrate approach, integrating with HRM and PCR for enhanced sensitivity and specificity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If ELISA-based methods are used for protein detection, then high specificity is achieved through antibodies, but extensive sample handling and need for highly trained staff significantly increase costs and complexity
Solution Approach 1:
The patent replaces the mechanical/chemical ELISA system with a surface plasmon resonance (SPR) optical sensing system. The SPR sensor detects molecular binding events through optical signal changes, eliminating the need for colorimetric or fluorescent readouts required in ELISA. This substitution reduces operational complexity while maintaining high specificity through the optical detection mechanism.
Solution Approach 2:
The SPR sensor platform provides a universal detection system that can analyze multiple analyte types (proteins, nucleic acids, small molecules) using the same core technology. The system uses a common SPR sensing mechanism with different bioreceptors immobilized on the sensor surface, eliminating the need for separate optimized assays for each analyte type, thus reducing overall complexity and cost.
2Measurement precision
If HRM-based methods are used for DNA detection, then higher sensitivity is achieved, but the methods are relatively complex, sensitive to contamination, and require thermocycling
Solution Approach 1:
The patent replaces the thermal cycling mechanism of HRM with an isothermal SPR detection system. The SPR sensor operates at constant temperature, detecting DNA binding events through real-time optical signal changes. This eliminates the complex thermocycling process while maintaining high sensitivity through the optical detection of molecular interactions.
Solution Approach 2:
The SPR sensor surface acts as an intermediary platform that captures DNA analytes through immobilized bioreceptors. This intermediary approach allows direct detection of DNA binding events without requiring amplification or thermal denaturation steps, simplifying the assay while maintaining sensitivity through the optical transduction mechanism.
3Measurement precision
If assay components are optimized for a certain group of molecules, then high performance is achieved for that specific target, but adaptation towards another target requires re-starting the entire assay development process
Solution Approach 1:
The SPR sensor platform provides a universal detection system where the core sensing mechanism remains constant while only the immobilized bioreceptor needs to be changed for different analytes. The same SPR instrumentation, data acquisition system, and analysis software can be used across multiple applications, enabling rapid adaptation to new targets without redeveloping the entire assay.
Solution Approach 2:
The assay system is segmented into modular components: a universal SPR sensing platform and interchangeable bioreceptor layers. This segmentation allows the performance-optimized detection system to remain constant while only the specific recognition element (bioreceptor) is changed for different analytes, facilitating rapid assay adaptation.
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 SPR sensor system provides a highly sensitive, cost-effective, and adaptable method for NAzyme activity analysis, capable of detecting different types of target molecules and monitoring molecular binding events in real-time, overcoming the limitations of existing techniques.
Implementation Method 1
surface plasmon resonance (SPR) sensor system that measures nucleic acid enzyme (NAzyme) activity
Implementation Method 2
metallic nanoparticle-labelled DNA sequences
Data Source
AI summary
NAzyme activity surface plasmon resonance sensors include a first DNA probe that is covalently connected to a sensing surface, and a second DNA probe that is covalently connected to a nanoparticle or a nanoparticle cluster. The first DNA probe and the second DNA probe are ligated together to provide a selected single strand DNA probe connected to the sensing surface and the nanoparticle. The single strand DNA probe includes a ligation zone within a selected NAzyme substrate. The sensor measures DNAzyme activity by NAzyme binding at the NAzyme substrate and cleavage at the ligation zone. Fiber optic surface plasmon resonance sensor tips are adapted to measure activity of a NAzyme when the NAzyme substrate is recognized by the selected NAzyme through hybridization and the metallic nanoparticle is released from the sensor by cleavage of the single strand DNA at the ligation zone by the selected NAzyme.