Immunoassay Bead Separation Without Washing
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Solution Overview
Problem
Current immunoassay methods for detecting target molecules in samples require multiple steps, including washing, which increases complexity, reagent usage, and time, making them unsuitable for rapid point-of-care diagnostics, especially in emergency situations like antimicrobial resistance and sepsis diagnosis.
Innovation Solution
A method involving the sequential steps of incubating a sample with capture and detection molecules coupled to beads in a buffer solution, separating the capture beads from the detection beads, and detecting the remaining detection molecules in the buffer solution without a washing step, allowing for simultaneous incubation, separation, and detection in the same reaction well.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If traditional immunoassay methods with washing steps are used, then specificity is improved, but the number of steps and time required increases
Solution Approach 1:
The invention extracts and eliminates the washing step from the traditional immunoassay protocol. By detecting unbound detection beads in the supernatant without washing, the method removes the time-consuming washing step while maintaining assay specificity through mathematical correction of background signal
Solution Approach 2:
Instead of detecting bound detection beads on the solid phase (traditional approach), the invention inverts the detection strategy by measuring unbound detection beads in the liquid supernatant. This inversion eliminates the need for washing while preserving specificity through background subtraction calculations
2Measurement precision
If washing steps are included, then signal-to-noise ratio is improved, but device complexity and reagent usage increase
Solution Approach 1:
The washing step is extracted and removed from the protocol. Mathematical correction methods are applied to calculate and subtract background signal from the measurement of unbound detection beads, achieving signal-to-noise ratio improvement without physical washing
Solution Approach 2:
Mathematical correction acts as an intermediary between the raw measurement of unbound detection beads and the final specific signal. The calculation process mediates the conversion of total signal (specific + background) into corrected specific signal
3Measurement precision
If multiple washing steps are performed, then detection accuracy is improved, but productivity decreases
Solution Approach 1:
Multiple washing steps are extracted and replaced by a single measurement of unbound detection beads in the supernatant, followed by mathematical correction. This dramatically reduces the number of operations while maintaining detection accuracy
Solution Approach 2:
The invention skips the washing steps entirely and rushes directly to detection of unbound beads in the supernatant. Mathematical correction rushes through the signal processing, achieving accurate results without the time-consuming washing operations
4Reliability
If washing steps are used, then reliability is improved, but ease of operation deteriorates
Solution Approach 1:
The washing step is extracted and removed, simplifying the操作流程 to incubation followed by direct detection. Mathematical correction automatically handles the reliability aspect without requiring manual washing operations
Solution Approach 2:
The system performs self-correction through mathematical calculations that automatically subtract background signal from total signal. This self-service approach maintains reliability without requiring operator-performed washing steps
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 reduces the number of steps and reagents needed, enhances sensitivity, and enables rapid, automated detection of target molecules, improving the efficiency and reliability of immunoassays for point-of-care applications by eliminating the need for washing and simplifying the assay process.
Implementation Method 1
separating in the reaction well the capture-beads from the buffer solution comprising unbound detection-beads
Implementation Method 2
wherein the capture-beads are separable from the detection-beads
Implementation Method 3
detection molecules suitable for binding to the target molecules and/or the capture molecules
Data Source
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AI summary
The invention relates to a method for detecting a target molecule in a sample, comprising the sequential steps of (a.) incubating at the same time in a buffer solution in a reaction well (i.) the sample suspected to contain one or more target molecules, (ii.) solid phase-coupled capture molecules suitable for binding to the target molecules, wherein the capture molecules are coupled to beads (capture-beads), (iii.) a defined amount of detection molecules suitable for binding to the target molecules and/or the capture molecules, wherein the detection molecules are coupled to beads (detection-beads), wherein the capture-beads are separable from the detection-beads, (b.) separating in the reaction well the capture-beads from the buffer solution comprising unbound detection-beads, and (c.) detecting the remaining unbound detection-beads in the buffer solution, which has been separated from the capture-beads, wherein the method does not comprise a wahing step.