Multi-epitope affinity assay concentration method
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Solution Overview
Problem
Affinity assays face challenges in achieving high signal-to-noise ratios and sensitivity due to non-specific binding and interference from endogenous components in biological samples, which limits their accuracy and reliability, especially in complex mixtures like blood or urine.
Innovation Solution
A method involving the use of capturing moieties adhered to particles that concentrate the biological target into a smaller volume, allowing for uncoupled binding and detection, thereby reducing interference and increasing specificity and sensitivity through enzymatic, thermal, or photochemical cleavage of the capturing moiety from the particle, enabling direct or indirect detection and quantification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional affinity assays are used to detect biological targets in complex biological samples, then the assay can identify the presence of targets, but the signal-to-noise ratio is low due to non-specific binding and interference from endogenous components
Solution Approach 1:
The assay is divided into multiple sequential steps: capture step with capturing moiety, washing step to remove interferents, elution step to release the target, and detection step. This segmentation allows separation of specific binding events from non-specific background, thereby improving signal-to-noise ratio.
Solution Approach 2:
The biological target is extracted from the complex biological sample matrix through specific binding to the capturing moiety. The target is then eluted from the capturing moiety into a clean eluate, removing it from interfering endogenous components. This extraction process isolates the target signal from background noise.
2Quantity of substance
If capturing moieties are adhered to particles for concentration, then the biological target can be concentrated into smaller volume, but the detecting and quantifying step becomes more complex
Solution Approach 1:
The capturing moiety acts as an intermediary that temporarily binds the biological target during concentration and purification steps. After elution, the capturing moiety is removed, allowing standard detection methods to be applied to the purified target in the eluate without interference from the capturing moiety or particle matrix.
Solution Approach 2:
The capturing moiety performs preliminary concentration and purification of the biological target before the detection step. By pre-concentrating the target from large sample volumes into small eluate volumes and removing interferents, the subsequent detection and quantification become simpler and more reliable.
3Reliability
If multiple capturing moieties are used to increase specificity, then the assay specificity improves, but the device complexity increases
Solution Approach 1:
Multiple capturing moieties recognizing different epitopes are used in a sequential manner across different steps (capture and detection), rather than simultaneously. This segmented approach maintains high specificity through multi-epitope recognition while keeping each individual step relatively simple and manageable.
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 signal-to-noise ratio and sensitivity of affinity assays, allowing for more accurate detection and quantification of biological targets with minimal false-positive results, suitable for point-of-care and miniaturized devices.
Implementation Method 1
adding a first capturing moiety to the biological sample volume comprising the biological target, wherein the first capturing moiety is adhered to a particle, c) concentrating the captured biological target into an elution volume that is smaller than the biological sample volume in step a)
Implementation Method 2
cleaving the first capturing moiety or the biological target from the particle
Implementation Method 3
cleaving the first capturing moiety or the biological target from the particle
Implementation Method 4
cleaving the first capturing moiety or the biological target from the particle
Data Source
Figure 1A~1D
Figure 2
Figure 3A~3D
AI summary
The present invention is related to a method for detection of a biological target in an affinity assay, the method comprising the steps of providing a biological sample volume containing the biological target, adding a first capturing moiety to the biological sample volume comprising the biological target, wherein the first capturing moiety is adhered to a particle, concentration of the captured biological target into an elution volume that is smaller than the biological sample volume in step a), cleavage of the first capturing moiety or the biological target from the particle and direct or indirect detection and/or quantification of the biological target in a sandwich or competitive affinity assay format, wherein the biological target is associated with at least one capturing moiety, preferably at least two capturing moieties.