Flow-Type SPR Biosensor for Real-Time Pathogen Detection
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Solution Overview
Problem
Conventional flow-type SPR sensors face challenges in detecting pathogenic microorganisms in real-time due to slow immune reaction speeds and weak detection signals, limiting their effectiveness in waterborne pathogen detection.
Innovation Solution
A modified flow-type SPR biosensor method is introduced, where the immune reaction between the pathogenic microorganism and antibody is performed in a batch type, followed by selective separation using a cut-off membrane filter, and the bound microorganism is then bound to a flow-type SPR chip, enhancing detection efficiency through biotin-streptavidin binding for increased SPR signal amplification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Speed
If a conventional flow-type SPR sensor is used for real-time detection, then the measurement speed is fast and results are obtained promptly, but the detection sensitivity and limit of detection are insufficient for pathogenic microorganisms
Solution Approach 1:
The detection process is divided into two separate stages: (1) batch-type immune reaction between pathogenic microorganisms and antibodies to form immune complexes, and (2) flow-type SPR detection of these complexes. This segmentation allows the slow immune reaction to occur in batch mode while maintaining the speed advantage of flow-type SPR for detection, resolving the contradiction between measurement speed and detection sensitivity.
Solution Approach 2:
The immune reaction between pathogenic microorganisms and antibodies is performed in advance in a batch-type process before introduction to the SPR sensor. This preliminary action concentrates the immune complexes and enhances the signal, thereby improving detection sensitivity while the subsequent flow-type SPR measurement maintains rapid detection capability.
2Measurement precision
If a batch-type sensor is used for pathogenic microorganism detection, then the detection sensitivity is improved, but the measurement time increases and real-time detection capability is lost
Solution Approach 1:
By separating the immune reaction step (batch-type, performed offline) from the detection step (flow-type, performed online), the method achieves high detection sensitivity through concentrated immune complexes while maintaining real-time detection capability during the flow-type SPR measurement phase.
Solution Approach 2:
The flow-type SPR detection continuously monitors the binding of immune complexes to the sensor surface in real-time, maintaining continuous useful action during the detection phase. This continuous monitoring provides real-time data without requiring prolonged incubation times associated with traditional batch-type sensors.
3Ease of operation
If a conventional flow-type SPR sensor directly detects pathogenic microorganisms, then the measurement is simple and label-free, but the immune reaction signal is weak and detection limit is high
Solution Approach 1:
The immune complexes are formed in advance in a batch-type process with optimized conditions for maximum binding efficiency. This preliminary action ensures that the complexes introduced to the SPR sensor have high binding affinity and concentration, thereby strengthening the detection signal while maintaining the simplicity of the flow-type SPR measurement process.
Solution Approach 2:
The immune complex acts as an intermediary between the pathogenic microorganism and the SPR sensor surface. By forming these complexes in batch mode with high concentration, the signal strength is enhanced when introduced to the sensor, improving detection sensitivity without complicating the overall measurement process.
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 significantly enhances the detection efficiency and sensitivity of pathogenic microorganisms, allowing for real-time detection of waterborne pathogens like cryptosporidium parvum, comparable to batch-type sensors, while maintaining the merits of flow-type SPR sensors, including non-labeled and prompt measurement.
Implementation Method 1
The surface plasmon is excited by light which passes a dielectric medium, such as a prism, and then which is incident upon a metal thin film at an angle being same as or greater than a critical angle of the dielectric medium. Then, the surface plasmon generates resonance at a predetermined angle. An angle of incidence at which the SPR is generated, that is, an angle of resonance, is sensitive to a change in the index of refraction of a material being close to the metal thin film.
Implementation Method 2
selectively separating the pathogenic microorganism bound with the antibody
Implementation Method 3
binding the pathogenic microorganism bound with the antibody on a chip of a flow-type SPR sensor system
Data Source
AI summary
There is provided a method of detecting a pathogenic microorganism in real-time, using a modified flow-type surface plasmon resonance (SPR) biosensor, comprising the steps of: i) performing, in a batch-type, an immune reaction of a pathogenic microorganism and an antibody thereto; ii) selectively separating the pathogenic microorganism bound with the antibody; and iii) binding the pathogenic microorganism bound with the antibody on a surface of a chip of a flow-type SPR sensor system in real-time.