DMP Tag/Anti-Tag Signal Amplification for Multiplex Immunoassays
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Solution Overview
Problem
Current bead-based multiplex immunoassays rely on biotin-streptavidin interaction for signal amplification, which may not provide sufficient amplification for detecting multiple analytes simultaneously and can be affected by background issues.
Innovation Solution
The use of a DMP-based chemical tag and a corresponding antibody that specifically binds to this tag, allowing for an alternative signal amplification method that can be used in conjunction with biotin-streptavidin without interference, thereby enhancing the ability to detect multiple analytes simultaneously.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If biotin-streptavidin system is used for signal amplification, then signal amplification is achieved, but background issues arise and amplification is insufficient for detecting multiple analytes simultaneously
Solution Approach 1:
The patent introduces a dimethyl piperidine (DMP) tag as an intermediary between the detection antibody and the fluorescent label. This DMP-based tag system acts as a mediator that replaces the problematic biotin-streptavidin interaction, providing signal amplification without the associated background issues. The DMP tag is conjugated to the detection antibody, and a fluorescently-labeled anti-DMP antibody provides the amplification signal, thereby resolving the contradiction between achieving amplification and avoiding background interference.
2Reliability
If biotin is conjugated to detection antibody, then signal amplification is possible, but multiple SA-PE binding partners attach causing interference in multiplex detection
Solution Approach 1:
The patent segments the signal amplification system into two independent pathways: one using biotin-streptavidin for the first analyte detection, and another using the DMP-based tag system for the second analyte detection. This segmentation allows each pathway to operate independently without interference, enabling true multiplex detection where multiple analytes can be detected simultaneously using different fluorescent channels without cross-reactivity or signal interference.
3Adaptability or versatility
If multiple detection antibodies are used for multiplex assay, then multiple analytes can be detected, but signal amplification becomes insufficient
Solution Approach 1:
The patent creates a composite detection system that combines two different amplification mechanisms: the biotin-streptavidin system and the DMP-based tag system. By using composite materials/concepts, the assay can simultaneously detect multiple analytes with sufficient signal amplification for each, as each analyte can be detected through its own dedicated amplification pathway, thereby resolving the contradiction between multiplex capability and signal amplification strength.
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 approach provides effective signal amplification, allowing for the simultaneous detection of multiple analytes with high specificity and sensitivity, while avoiding background issues associated with other systems.
Implementation Method 1
a first detection reagent capable of specifically binding to the first detectable label and including a first fluorescent label; and a second detection reagent capable of specifically binding to the second detectable label and including a second fluorescent label
Data Source
AI summary
Methods and kits for detecting analytes (such as a plurality of analytes) in a sample utilizing signal amplification (such as DMP-based chemical tag signal amplification) are provided.