Solid-Phase Binding Assay Analyte Pre-concentration
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Solution Overview
Problem
Existing binding assays face challenges in improving signal detection and measurement accuracy, especially when analyzing complex biological samples, due to interference from extraneous materials and low analyte concentrations.
Innovation Solution
The method involves a series of steps including contacting a sample with a particle or solid phase linked to a binding reagent, separating unbound components, releasing the complex, and measuring the analyte bound to a solid phase using a second binding reagent, which can include detectable labels for enhanced detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If binding assays are conducted directly on complex biological samples, then the assay can be performed with minimal processing, but the signal detection and measurement accuracy are reduced due to interference from extraneous materials and low analyte concentrations
Solution Approach 1:
The assay is divided into distinct stages: (1) capture of analyte-bound complexes on magnetic particles, (2) separation and washing to remove interferents, (3) release of complexes, and (4) detection on a solid phase. This segmentation allows removal of extraneous materials between steps, improving measurement precision while maintaining operational clarity through standardized protocols
Solution Approach 2:
Magnetic particles coated with capture reagents serve as intermediaries to concentrate analytes from complex samples. These particles facilitate the separation of analyte-bound complexes from extraneous materials through magnetic separation, enabling subsequent detection with improved accuracy without requiring direct analysis of the original complex sample
2Quantity of substance
If analyte concentration in complex biological samples is low, then the sample requires minimal preparation, but the signal obtained from binding events is insufficient for accurate measurement
Solution Approach 1:
The method performs preliminary concentration of analytes by capturing them on magnetic particles before the detection step. This pre-concentration increases the effective analyte concentration and signal strength for subsequent measurement, improving measurement precision without altering the fundamental binding assay methodology
Solution Approach 2:
The detection step merges the captured analyte complexes with a solid phase and detection reagents, concentrating the signal in a localized area. This merging of analyte, capture reagent, and detection reagent on a solid phase amplifies the measurable signal from low-abundance analytes in the original sample
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 enhances assay performance by pre-concentrating analytes and reducing contaminants, leading to improved signal detection and measurement accuracy in binding assays.
Implementation Method 1
techniques that employ binding reactions, e.g., antigen-antibody reactions
Implementation Method 2
nucleic acid hybridization
Implementation Method 3
receptor-ligand reactions
Implementation Method 4
The particle may be a magnetic particle and the collection step may comprise magnetic concentration of the particle
Implementation Method 5
In one embodiment, the detectable label is an electrochemiluminescent label
Data Source
AI summary
The invention relates to methods for conducting solid-phase binding assays. One example is an assay method having improved analyte specificity where specificity is limited by the presence of non-specific binding interactions.