Immunoassay Dye Detection to Minimize Hook Effect Interference
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Solution Overview
Problem
Immunoassays experience 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 in the immunoassay, allowing for simultaneous detection with an immunoassay reagent, to generate a signal and shift an absorption band, thereby minimizing hook effect interference.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a wash step is added between incubation steps to eliminate the hook effect, then measurement precision is improved, but device complexity and time consumption increase
Solution Approach 1:
The patent extracts the harmful wash step from the assay procedure by using a one-step mixing approach where capture antibody, detection antibody, and sample are combined simultaneously without intermediate washing, thereby eliminating the complexity while maintaining accuracy through the dye-based detection method
Solution Approach 2:
The patent applies preliminary action by pre-mixing capture antibody and detection antibody in a single reagent solution before adding the sample, allowing both antibodies to bind simultaneously to the analyte without requiring sequential washing steps that会增加 complexity
2Measurement precision
If multiple assays are conducted on a single sample to ensure validity, then measurement precision is improved, but productivity decreases
Solution Approach 1:
The patent makes the single assay universally reliable by incorporating a dye that binds to the analyte in addition to the immunoassay reagent, allowing the same assay to accurately measure both low and high analyte concentrations without requiring multiple separate assays, thereby maintaining productivity
Solution Approach 2:
The patent changes the detection parameter by using dye absorption spectroscopy alongside immunoassay signal detection, creating a dual-parameter system that can distinguish between true low signals and hook effect artifacts, ensuring assay validity in a single run
3Measurement precision
If antibody concentration is increased to handle high analyte concentrations, then measurement precision is improved, but cost increases
Solution Approach 1:
The patent introduces a dye as an intermediary substance that binds to the analyte and provides an additional detection signal. This dye acts as a mediator that works alongside the antibodies, allowing the system to handle high analyte concentrations without needing to increase antibody amounts, thus controlling reagent costs
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 extends the dose response range and reduces false negative results, providing accurate analyte concentration measurements without the need for additional wash steps, suitable for high-throughput analysis.
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.