Multiplex Binding Assays Using Bacteriocin-Immunity Protein Couples
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Solution Overview
Problem
Current multiplex binding assays face challenges with sensitivity and specificity due to cross-reactions among analytes, particularly in lateral flow biosensors, which affect the accuracy and reliability of simultaneous detection of multiple analytes.
Innovation Solution
The use of polypeptide couples with high affinity, specifically bacteriocins and their cognate immunity proteins, functionalized on a support with low dissociation constants in the picomolar or femtomolar range, to enhance specificity and sensitivity in multiplex binding experiments.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If conventional antibodies and antigens are used in multiplex binding assays, then the assays can detect multiple analytes simultaneously, but cross-reactions occur among analytes reducing sensitivity and specificity
Solution Approach 1:
The patent changes the binding parameter (dissociation constant) from nanomolar range (conventional antibodies) to picomolar/femtomolar range (bacteriocin-immunity protein couples). This parameter change enables highly specific binding that prevents cross-reactions while maintaining multiplex detection capability
Solution Approach 2:
The patent uses composite binding systems comprising bacteriocin polypeptides coupled to capture entities and immunity polypeptides as detection entities. This composite approach combines the high affinity of bacteriocins with the specificity of immunity proteins to eliminate cross-reactivity in multiplex assays
2Productivity
If lateral flow biosensors are used for multiplex testing, then diagnostic time and costs are reduced, but sensitivity and specificity problems arise due to cross-reactions
Solution Approach 1:
The patent applies parameter changes by using bacteriocin-immunity protein couples with picomolar/femtomolar dissociation constants in lateral flow biosensors. This enables the maintenance of diagnostic efficiency while achieving high reliability through eliminated cross-reactions
Solution Approach 2:
The immunity polypeptides act as intermediaries that specifically recognize and bind only to their cognate bacteriocins. This intermediary mechanism ensures that detection entities only bind to intended capture entities, preventing cross-reactions while maintaining assay efficiency
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 significantly reduces cross-reactivity among non-cognate pairs, allowing for precise and sensitive detection of multiple analytes in a single run, improving the reliability and efficiency of multiplex assays.
Implementation Method 1
different polypeptide couples having high affinity, able to display molecules of interest in an appropriate manner on the surface of a support
Implementation Method 2
The polypeptides are coupled to the support by any suitable method, in particular by non-covalent interactions
Implementation Method 3
a labeled detection entity, which is an antibody or an antigen, without disturbing its binding to the analyte-capture entity
Data Source
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AI summary
The present invention concerns a support for a multiplex binding experiment functionalized with at least two different polypeptides having high affinity to their cognate binding partners provided in a reaction mixture, and their use in e.g. immunoassays.