New Hapten Classes for Multiplexed Immunoassay Sensitivity
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Solution Overview
Problem
There is a lack of research on developing additional classes of haptens against which sensitive and specific antibodies can be raised for highly multiplexed assays, which are essential for monitoring therapeutic responses and companion diagnostic applications.
Innovation Solution
The development of new classes of haptens, such as oxazoles, pyrazoles, thiazoles, nitroaryls, benzofurazans, triterpenes, ureas, thioureas, rotenones, coumarins, and cyclolignans, along with their conjugates and anti-hapten antibody conjugates, for use in enzyme immunoassays and diagnostic methods to detect multiple targets in biological samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional haptens (biotin, fluorescein) are used for immunoassays, then the assay can be performed, but background interference occurs due to endogenous biotin or background fluorescence
Solution Approach 1:
The patent introduces digoxigenin as an intermediary hapten that does not naturally occur in biological systems. This mediator allows specific binding through anti-digoxigenin antibodies without the background interference problems associated with biotin (endogenous biotin) or fluorescein (background fluorescence), thereby resolving the contradiction between assay sensitivity and background interference
Solution Approach 2:
The patent employs biotin-streptavidin-biotinylated anti-digoxigenin antibody conjugates as a disposable detection system. The biotinylated conjugates are used once for detection and then discarded, avoiding the need for repeated use that would accumulate background interference, while maintaining high sensitivity through the strong biotin-streptavidin interaction
2Productivity
If only a limited number of hapten classes are available, then current immunoassays can function, but multiplexed assays for simultaneous detection of multiple targets are limited
Solution Approach 1:
The patent creates universal detection platforms using biotinylated anti-digoxigenin antibody conjugates that can detect multiple different targets (proteins, nucleic acids) by simply changing the hapten-labeled primary antibody, while maintaining a constant secondary detection system. This multi-functional approach enables multiplexed assays without requiring diverse hapten classes
Solution Approach 2:
The patent segments the immunoassay into distinct functional modules: (1) target-specific primary antibodies labeled with various haptens, (2) universal biotinylated anti-digoxigenin antibody conjugates, and (3) streptavidin-based signal detection. This segmentation allows flexible combination for multiplexing while maintaining system simplicity
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
These new haptens and conjugates enable advanced multiplexed assays, allowing for the simultaneous detection and quantitation of multiple protein and nucleic acid targets, enhancing the sensitivity and specificity of diagnostic and therapeutic applications.
Implementation Method 1
specific binding of antidigoxin antibodies to the drug in the test sample
Implementation Method 2
an enzyme or a fluorescent label
Implementation Method 3
a fluorescent label
Data Source
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AI summary
A method for performing a multiplexed diagnostic assay, such as for two or more different targets in a sample, is described. One embodiment comprised contacting the sample with two or more specific binding moieties that bind specifically to two or more different targets. The two or more specific binding moieties are conjugated to different haptens, and at least one of the haptens is an oxazole, a pyrazole, a thiazole, a nitroaryl compound other than dinitrophenyl, a benzofurazan, a triterpene, a urea, a thiourea, a rotenoid, a coumarin, a cyclolignan, a heterobiaryl, an azo aryl, or a benzodiazepine. The sample is contacted with two or more different anti-hapten antibodies that can be detected separately. The two or more different anti-hapten antibodies may be conjugated to different detectable labels.