Lipid-Layer Immunosensing with Stable Sandwich Complexes
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Solution Overview
Problem
Existing ELISA methods face challenges with sensitivity and specificity due to fragile immune complexes, leading to false positive signals and limitations in signal generation, particularly in sandwich formats.
Innovation Solution
A method involving a sensor element with an anchor layer, a first binding agent anchored in the layer and comprising a detectable label, and a second binding agent immobilized on a solid support, allowing for specific binding and detection of analytes through a 'lock-on, lock-off' complex formation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If traditional ELISA methods are used with immobilized capture agents and detection agents, then the assay can detect analytes, but the immune complexes are fragile leading to false positive signals and reduced sensitivity
Solution Approach 1:
The patent applies preliminary action by pre-forming stable immune complexes on a microbead support before introducing them to the assay system. The capture agent is pre-bound to the analyte and detection agent complex on the microbead, creating a stable sandwich structure that prevents false positives while maintaining sensitivity. This pre-assembly of the complex eliminates the fragility issues of traditional ELISA where complexes form and break during multiple washing steps.
2Reliability
If multiple washing steps are performed in traditional ELISA to reduce background noise, then specificity improves, but the fragile immune complexes may dissociate leading to false negatives
Solution Approach 1:
The immune complex is pre-assembled on the microbead support with the capture agent, analyte, and detection agent already bound in a stable sandwich configuration. This preliminary formation of the complete complex on the stable microbead support eliminates the need for multiple washing steps that could cause dissociation, as the complex remains intact throughout the assay process.
3Measurement precision
If sandwich format ELISA is used for signal generation, then sensitivity can be improved through enzyme amplification, but the complexity of multiple reagent additions and washing steps increases
Solution Approach 1:
The patent merges the capture agent, detection agent, and analyte into a single pre-formed sandwich complex on the microbead support. This consolidation eliminates the need for separate addition and washing steps for each reagent, simplifying the assay procedure while maintaining the signal amplification benefits of the sandwich format through the enzyme-linked detection agent.
Solution Approach 2:
The complete sandwich complex is pre-assembled before the assay begins, with the capture agent bound to the analyte-detection agent complex on the microbead. This preliminary formation of the complete immunocomplex structure eliminates the need for multiple sequential reagent additions and washing steps, reducing procedural complexity while preserving the sensitivity advantages of the sandwich format.
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
Improves sensitivity and specificity by reducing background noise and enabling dynamic, real-time detection of analytes with improved accuracy, potentially eliminating the need for washing steps.
Implementation Method 1
a first binding agent which is capable of specifically binding to the analyte
Implementation Method 2
a second binding agent which is capable of specifically binding to the analyte when bound to the first binding agent
Data Source
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AI summary
The present invention relates to diagnostic test and technology. In particular, it relates to the present invention relates to a method for determining an analyte suspected to be present in a sample comprising contacting said sample with a sensor element comprising i) an anchor layer which is present on a solid support, ii) a first binding agent which is capable of specifically binding to the analyte, which is anchored in the anchor layer and which comprises at least one detectable label, and iii) a second binding agent which is capable of specifically binding to the analyte when bound to the first binding agent and which is immobilized on the solid support, for a time and under conditions which allow for specific binding of the analyte suspected to be present in the sample to the first binding agent and specific binding of the second binding agent to the analyte bound to the first binding agent and detecting the formation of the complex of first binding agent, analyte and second binding agent whereby the analyte is determined. Moreover, provided is a device for determining an analyte suspected to be present in a sample and the use thereof for determining an analyte suspected to be present in a sample in said sample. Moreover, the present invention contemplates a kit for determining an analyte suspected to be present in a sample.