Dye-Shift Immunoassay Detection for High Analyte Concentrations
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Solution Overview
Problem
Immunoassays suffer from the 'hook effect' due to high analyte concentrations saturating antibodies, leading to false negative or false low concentration results, which current compensation methods like calibration curves and wash steps increase assay time and complexity.
Innovation Solution
Incorporating a dye that binds to the target analyte and shifts its absorption band upon binding, combined with an immunoassay reagent, to detect the analyte and minimize hook effect interference.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a wash step is added between capture antibody incubation and detection antibody addition to eliminate the hook effect, then measurement precision is improved, but loss of time and device complexity increase
Solution Approach 1:
The patent extracts the harmful effect (hook effect) by adding a detergent solution that selectively removes excess analyte from the capture antibody-bound complex, while leaving the properly formed immune complexes intact. This extraction of the interfering substance eliminates the need for a wash step while maintaining measurement precision.
Solution Approach 2:
The detergent solution acts as an intermediary substance that mediates between the capture antibody-analyte complex and the detection antibody. It selectively interacts with excess analyte to remove it, facilitating accurate detection without requiring time-consuming wash steps.
2Measurement precision
If multiple assays are conducted on a single sample to ensure validity and compensate for the hook effect, then measurement precision is improved, but productivity decreases
Solution Approach 1:
The patent performs a preliminary action by adding the detergent solution to remove excess analyte before the detection antibody is introduced. This preliminary removal of interfering substances ensures that a single assay run provides valid results, eliminating the need for multiple assays and thereby maintaining high productivity.
3Measurement precision
If the concentration of antibody reagents is increased to handle high analyte concentrations, then measurement precision is improved, but device complexity and cost increase
Solution Approach 1:
The patent changes the chemical parameter of the assay environment by introducing a detergent solution with specific surfactant properties. This parameter change allows the assay to handle high analyte concentrations without increasing antibody reagent concentrations, thereby maintaining simplicity in assay design while improving measurement precision.
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
The method allows for accurate detection of high analyte concentrations without saturation, reducing false results and maintaining assay efficiency by extending the dose response curve.
Implementation Method 1
at least one dye capable of binding to the target analyte, wherein an absorption band of the at least one dye shifts upon binding to the target analyte
Implementation Method 2
at least one immunoassay reagent capable of specifically binding to the target analyte and capable of directly or indirectly generating a signal upon binding to the target analyte
Data Source
AI summary
Methods of minimizing hook effect interference in an immunoassay are disclosed. Also disclosed are reagents, kits, and immunoassay devices that may be utilized in accordance with the method.