Dual-Label Immunoassay Quantitation Across Low and High Concentrations
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Solution Overview
Problem
Existing detection methods using fluorescent and chemiluminescent labels face challenges with nonlinear relationships and inaccuracies due to self-quenching at high concentrations and inefficiencies at low concentrations, leading to unreliable quantitation.
Innovation Solution
A dual-label detection method utilizing both fluorescence and chemiluminescence signals, where fluorescence is used for low concentrations and chemiluminescence for high concentrations, with a cutoff value determining which calibration curve to use for accurate quantitation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If fluorescent label is used for detection, then low concentration targets can be effectively detected, but at high concentrations self-quenching occurs leading to nonlinear relationship and inaccurate quantitation
Solution Approach 1:
The patent changes the detection parameter by switching between fluorescence and chemiluminescence signals based on analyte concentration. At low concentrations, fluorescence signal is used for high sensitivity detection. At high concentrations, chemiluminescence signal is used to avoid self-quenching effects, thereby maintaining quantitation reliability across the full dynamic range.
Solution Approach 2:
The detection range is segmented into low concentration and high concentration zones. Different detection modalities (fluorescence vs. chemiluminescence) are applied to different segments of the concentration range, with each modality optimized for its appropriate concentration zone to ensure accurate quantitation throughout.
2Measurement precision
If chemiluminescent label is used for detection, then high concentration samples can be accurately detected, but at low concentrations assay variation increases
Solution Approach 1:
The detection parameter is dynamically changed based on analyte concentration level. Chemiluminescence signal is utilized for high concentration samples where it provides accurate detection, while fluorescence signal is employed for low concentration samples where it maintains higher reliability and lower assay variation.
Solution Approach 2:
The concentration range is divided into segments with different optimal detection methods. The chemiluminescence channel handles the high concentration segment, while the fluorescence channel handles the low concentration segment, maximizing the strengths of each detection modality.
3Device complexity
If only fluorescent label is used, then detection system is simple, but quantitation becomes inaccurate at high concentrations due to self-quenching
Solution Approach 1:
The patent merges two detection systems (fluorescence and chemiluminescence) into a single immunoassay platform. Both labeling modalities are combined on the same detection surface, allowing the system to leverage the advantages of each: fluorescence for low concentration sensitivity and chemiluminescence for high concentration linearity, thereby improving overall quantitation accuracy.
Solution Approach 2:
The detection system is designed with multi-functionality to perform both fluorescence and chemiluminescence detection. This universal platform can adaptively switch between detection modes based on analyte concentration, providing accurate quantitation across the entire dynamic range without requiring separate assay systems.
4Device complexity
If only chemiluminescent label is used, then detection system is simple, but assay variation increases at low concentrations
Solution Approach 1:
Two detection modalities (fluorescence and chemiluminescence) are merged into a single assay system. The fluorescence channel is specifically optimized for low concentration detection with higher reliability and lower variation, while the chemiluminescence channel handles higher concentrations, together providing robust performance across the full range.
Solution Approach 2:
The immunoassay platform is designed with universal capability to perform both fluorescence and chemiluminescence detection using the same capture and detection antibody pairs. This multi-functional system allows selective use of the optimal detection modality for each concentration range, improving low concentration assay reliability.
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 provides accurate quantitation across a wide range of analyte concentrations by leveraging the strengths of both fluorescent and chemiluminescent signals, ensuring high sensitivity and precision in both low and high concentration measurements.
Implementation Method 1
fluorescent molecules are commonly used... Although a fluorescent label can effectively detect targets with a low-end concentration
Implementation Method 2
Chemiluminescent label requires no external radiant energy for detection. The energy comes from photons released after the chemical reaction breaks the chemical bond.
Data Source
AI summary
The present invention is directed to an immunoassay method for detecting an analyte in a liquid sample; and provides accuracy and reproducibility for both high concentration and low concentration samples. The method uses both fluorescence marker and chemiluminescence marker, and reads both fluorescence signal and chemiluminescence signal. The analyte concentration is determined based on either a calibration curve for the fluorescence signal, or a calibration curve chemiluminescence signal, using a pre-established analyte concentration value.