Fluorescence Quenching Immunoassay for Separation-Free Detection
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Solution Overview
Problem
Current immunoassays lack sensitivity, are complex to perform, and are costly, necessitating the development of methods that can accurately determine analyte concentrations with enhanced sensitivity and simplicity.
Innovation Solution
The method involves contacting a sample with a fluorescent tracer and a binding partner specific to the analyte, then irradiating the mixture with light at a specific wavelength to measure the intensity of emitted light, which is proportional to the analyte concentration.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional immunoassay methods are used, then analyte concentration can be determined, but the sensitivity is insufficient and the assay is complex to perform
Solution Approach 1:
The patent employs fluorescence quenching parameters to detect analyte concentration. By measuring changes in fluorescence intensity (a physical parameter) when the fluorescent tracer binds to the binding partner, the assay achieves high sensitivity without complex separation steps. The fluorescent tracer's emission properties change upon binding, providing a direct readout of analyte concentration.
Solution Approach 2:
The patent replaces mechanical separation steps with optical detection. Instead of physically separating bound from free tracer through washing and filtration steps, the assay uses fluorescence quenching measurements to directly detect binding events. This substitution of mechanical operations with optical measurement simplifies the assay procedure while maintaining measurement precision.
2Measurement precision
If conventional immunoassay methods are used, then analyte concentration can be determined, but the assay is costly
Solution Approach 1:
The patent uses fluorescent tracers that are chemically modified derivatives of fluorescein, which are relatively inexpensive fluorescent compounds. These tracers can be synthesized through standard chemical modification procedures, making the assay cost-effective compared to using expensive radioactive labels or complex enzyme systems. The tracers are designed for single-use in each assay.
3Measurement precision
If separation steps are included in the immunoassay, then binding events can be accurately measured, but the assay efficiency decreases
Solution Approach 1:
The patent enables continuous measurement of binding events without interruption for separation steps. The fluorescent tracer remains in solution throughout the assay, and binding can be monitored continuously by measuring fluorescence quenching. This eliminates the need to stop the reaction, separate components, and restart, thereby maintaining continuous useful action and improving assay efficiency.
Solution Approach 2:
The patent extracts the separation step from the assay procedure entirely. By using fluorescence quenching as the detection mechanism, the assay can directly measure binding events in the presence of both bound and free tracer without requiring physical separation. This extraction of the separation step simplifies the workflow and improves productivity.
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 allows for the sensitive and accurate determination of analyte concentrations without the need for separation steps, thereby improving assay efficiency and reducing costs.
Implementation Method 1
Fluorescence quenching immunoassay for determining the concentration of an analyte in a sample
Data Source
AI summary
The present invention relates to a method and reagents for determining the presence of or the amount of an analyte in a sample.