Multiplex ELISA Solid Substrate Analyte Detection
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Solution Overview
Problem
Current methods for detecting multiple analytes in a single sample face challenges such as distinguishing between analytes, overlap and interference of detection signals, insufficient sensitivity, and interference from other agents, particularly in complex biological samples, and often require expensive equipment or new technologies.
Innovation Solution
A method involving a solid substrate with immobilization agents, multiple antibody capture agents, and detectable agents that bind to different analytes, allowing for simultaneous detection by washing and identifying bound agents, which can include peptide-tag and anti-peptide antibody pairs, lanthanide ions, or enzyme substrates, to enhance sensitivity and reduce interference.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple analytes are detected in parallel using separate reactions, then the ability to distinguish between analytes is improved, but the time consumption and resource usage increase
Solution Approach 1:
The patent combines multiple separate analyte detection reactions into a single reaction well by using a multiplex ELISA approach. Different capture antibodies specific to various analytes are immobilized on the solid substrate, allowing simultaneous detection of multiple analytes in one reaction, thereby reducing time consumption while maintaining the ability to distinguish between different analytes through specific antibody recognition
Solution Approach 2:
The solid substrate serves multiple functions simultaneously: it acts as a platform for immobilizing multiple different capture antibodies, provides a common detection surface for multiple analytes, and enables single-well reaction conditions. This multi-functional design allows one reaction system to perform what would traditionally require multiple separate reactions
2Productivity
If multiple analytes are detected in the same reaction using multiplex detection, then the time efficiency is improved, but signal overlap and interference between detection signals worsen
Solution Approach 1:
Each capture antibody on the solid substrate is specific to a particular analyte, creating locally distinct binding sites that prevent cross-reactivity. The detection antibodies are also analyte-specific, ensuring that each analyte produces a unique detectable signal. This local specificity maintains signal distinction even when multiple analytes are detected simultaneously in the same reaction well
3Productivity
If traditional multiplex detection methods are used, then multiple analytes can be detected in a single reaction, but the sensitivity to detect low amounts of analyte deteriorates
Solution Approach 1:
The patent uses detection antibodies that are conjugated to detectable agents (such as enzymes, fluorophores, or other detectable molecules) to amplify and copy the signal from each analyte. This signal amplification mechanism allows sensitive detection of low analyte concentrations while maintaining the multiplex capability to detect multiple analytes simultaneously in the same reaction
4Measurement precision
If new technologies and equipment are adopted for multiplex detection, then the detection capability is improved, but the cost and device complexity increase
Solution Approach 1:
The patent employs a solid substrate-based ELISA format that can be performed using standard, readily available laboratory equipment and reagents. The method utilizes conventional plate readers and standard detection chemistry rather than requiring expensive specialized instruments, making the multiplex detection capability accessible to laboratories with existing resources
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 enables efficient detection of multiple analytes in a single reaction environment with improved sensitivity, reduced interference, and cost-effectiveness, utilizing existing hardware like plate readers, and can handle complex biological samples with moderate to high protein concentrations.
Implementation Method 1
Detection of analytes in such samples often utilises recognition of the analyte by binding to another molecule
Implementation Method 2
the detectable agents that bind to the at least two different analytes; wherein one or more of the at least two different detectable agents comprises a lanthanide ion
Implementation Method 3
the detectable agents that bind to the at least two different analytes; wherein one or more of the at least two different detectable agents comprises an enzyme that converts a substrate into a detectable product
Data Source
Figure 1A~1B
Figure 2A~2B
Figure 3
AI summary
The present disclosure relates to methods and kits for detecting multiple analytes in a sample. Certain embodiments are directed to a method for detecting different analytes in a sample comprising providing at least two different capture agents that bind to at least two different analytes and the at least two different capture agents can be immobilised on at least one solid substrate. Certain embodiments are directed to the use of pre-immobilised capture agents. Certain other embodiments are directed to capture agents that are immobilised on a solid substrate using an affinity capture system, including a peptide tag/anti-peptide tag antibody affinity capture system.