Fluorescent Competition Assay for Rapid Small Molecule Detection
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Solution Overview
Problem
Current methods for detecting small molecule analytes in blood samples are often time-consuming, require specialized equipment, and are not suitable for point-of-care (POC) applications, especially for drugs where enzymatic conversion is not available.
Innovation Solution
A fluorescently based competition assay using optically labelled analyte binding proteins and analogues, where the analyte binding protein is covalently linked to a first member of an optical signal pair and the analyte analogue is linked to a second member, allowing for rapid determination of analyte presence and concentration by measuring optical signals.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If HPLC-MS is used for analyte detection, then measurement precision is improved, but productivity deteriorates due to time-consuming procedures and requirement for specialists
Solution Approach 1:
The patent replaces the mechanical/chemical HPLC-MS system with an optical detection system using fluorescently labelled analyte binding proteins. The binding interaction between analyte and binding protein is detected through optical signals (fluorescence), eliminating the need for complex mechanical chromatography and mass spectrometry equipment, thereby enabling rapid point-of-care detection while maintaining measurement precision
Solution Approach 2:
The patent uses fluorescently labelled copies (analogs) of the analyte that bind to the analyte binding protein with similar affinity. These labelled copies serve as detectable proxies for the actual analyte, allowing optical detection of analyte concentration through competitive binding assays without requiring direct detection of the original analyte molecules
2Productivity
If enzymatic conversion is used for glucose detection, then productivity is improved, but adaptability deteriorates since it only works for glucose
Solution Approach 1:
The patent creates a universal detection platform based on analyte binding proteins that can detect multiple different analytes (glucose, drugs, metabolites) by simply changing the binding protein component. The same optical detection system and assay format can be applied to various analytes, making the system multi-functional and adaptable beyond just glucose detection
Solution Approach 2:
The patent introduces analyte binding proteins as intermediary molecules that mediate between the target analyte and the optical detection system. These binding proteins specifically recognize and bind various analytes, converting diverse chemical structures into a common binding event that can be detected optically, thereby enabling versatile detection of different analyte types
3Measurement precision
If Surface Plasmon Resonance is used for small molecule detection, then measurement precision is improved, but device complexity increases due to expensive equipment requirements
Solution Approach 1:
The patent employs relatively simple, inexpensive optical detection equipment and disposable fluorescently labelled binding proteins instead of expensive, complex SPR instrumentation. The fluorescent assay reagents can be prepared and used in simple cuvettes or microplates with basic fluorescence readers, replacing the need for sophisticated SPR instruments while achieving comparable detection precision
Solution Approach 2:
The patent replaces the complex SPR optical system (requiring specialized lasers, prisms, and detectors) with a simpler fluorescence detection system. The binding event is detected through fluorescence signal changes rather than SPR angle shifts, eliminating the need for complex optical path configuration and specialized SPR equipment
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 method enables fast and reliable detection of analytes in blood samples, including complex matrices like plasma and whole blood, with results obtainable within minutes, improving upon existing technologies by providing a sensitive and efficient means for POC analysis.
Implementation Method 1
a fluorescently based competition assay comprising a fluorescently labelled analyte binding protein and a fluorescently labelled analyte analogue
Data Source
AI summary
The present invention relates to an in vitro method for determining the presence, absence and/or concentration of an analyte in a sample. The method uses an optically based competition assay comprising a labelled analyte binding protein and a labelled analyte analogue. The concentration/presence of the analyte is determined by inhibitory binding of the analyte to the analyte binding protein thereby impeding binding of the analyte analogue to the analyte binding protein. The invention further relates to kits, solid supports, cartridges, detection chips and uses thereof.