Single-Molecule Capture-and-Release Detection System
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Solution Overview
Problem
Current methods for detecting low concentrations of analyte molecules in samples are limited by ensemble responses requiring large numbers of molecules, non-specific binding, and high background signals, which restrict sensitivity and dynamic range.
Innovation Solution
A method involving a capture-and-release system where analyte molecules associate with capture components on a substrate, are dissociated, and partitioned across reaction vessels for detection, allowing for the determination of low concentrations with reduced non-specific binding and improved sensitivity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If amplification procedures (ELISA, PCR, immunoPCR) are used to increase reporter molecules, then detection sensitivity is improved, but assay complexity increases and false-positive signals are generated
Solution Approach 1:
The patent extracts and eliminates the amplification step from the detection system. Instead of using ELISA, PCR, or immunoPCR to amplify signals, the invention directly detects individual analyte molecules bound to capture components on a substrate, removing the source of false positives and complexity while maintaining sensitivity through single-molecule detection capabilities
Solution Approach 2:
The patent uses fluorescently labeled capture components that act as copies or proxies for the analyte molecules. These labeled capture components bind to the analytes and provide detectable signals without requiring amplification, allowing direct quantification of individual molecule binding events
2Quantity of substance
If ensemble response methods are used requiring large numbers of molecules, then detection threshold is reached, but sensitivity and dynamic range are limited
Solution Approach 1:
The patent inverts the traditional ensemble detection approach by transitioning to single-molecule detection. Instead of requiring large numbers of molecules to generate a measurable signal, the system detects individual analyte molecules bound to capture components, achieving high sensitivity and extended dynamic range by counting discrete binding events
Solution Approach 2:
The patent replaces the mechanical/physical ensemble averaging approach with a molecular-level detection mechanism. By using fluorescently labeled capture components and single-molecule imaging techniques, the system substitutes bulk measurements with individual molecule detection, enabling sensitivity beyond traditional ensemble methods
3Object-generated harmful factors
If non-specific binding occurs to sites other than expected, then background signal increases, but detection accuracy decreases
Solution Approach 1:
The patent applies local quality by using specifically designed capture components with high affinity for the target analyte. The capture components are localized to specific binding sites on the substrate and are optimized to recognize only the target molecule, minimizing non-specific binding through careful selection of binding partners and controlled local environment
Solution Approach 2:
The patent introduces fluorescently labeled capture components as intermediaries between the analyte and the detection system. These intermediaries provide specific binding to the analyte while their fluorescent labels provide the detection signal, separating the binding function from the detection function and reducing non-specific interactions
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
Enables the detection and quantification of analyte molecules at lower concentrations with enhanced sensitivity and reduced false positives, facilitating the detection of molecules previously undetectable due to low concentrations.
Implementation Method 1
exposing a substrate comprising a plurality of capture components to a sample comprising a plurality of analyte molecules or particles, so that analyte molecules or particles associate with capture components to form a plurality of complexes
Implementation Method 2
dissociating at least a portion of each complex to form a plurality of dissociated species, which are not immobilized with respect to the substrate
Data Source
AI summary
The present invention relates to systems and methods for detecting analyte molecules or particles in a fluid sample and in some cases, determining a measure of the concentration of the molecules or particles in the fluid sample. Methods of the present invention may comprise immobilizing a plurality of analyte molecules or particles to form a plurality of complexes, releasing at least a portion of some of the plurality of complexes, determining at least a portion of the plurality of complexes released, and determining a measure of the concentration of the analyte molecules or particles in a fluid sample.