Split Affinity Marker for Multiplex Proximity Detection
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Solution Overview
Problem
Existing markers for biological samples lack flexibility and reliability, particularly in detecting multiple target analytes with high specificity and reducing false-positive rates, especially in multiplex assays and diagnostic tests.
Innovation Solution
A marker comprising two parts with specific affinity reagents and polymeric backbones, where labelling moieties form upon proximity, allowing optical detection of target analytes, and can be controlled through catalytic conditions to enhance specificity and reduce false positives.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If traditional markers with single affinity reagents and labels are used, then the marker structure is simple, but the flexibility and reliability in detecting multiple target analytes is insufficient
Solution Approach 1:
The marker is divided into two separate marker parts: a first marker part with a first affinity reagent and first labelling moiety parts, and a second marker part with a second affinity reagent and second labelling moiety parts. This segmentation allows each part to independently bind to different target analytes or different epitopes, enabling flexible multiplex detection while maintaining manageable complexity through modular design
Solution Approach 2:
The marker system is designed to perform multiple functions: it can detect different target analytes simultaneously, determine proximity between targets, and provide high-specificity detection through dual-affinity reagent binding. The universal design allows the same marker structure to be applied across various diagnostic and research applications
2Productivity
If traditional markers are used for multiplex assays, then the assay throughput is high, but the false-positive rate increases due to cross-reactivity
Solution Approach 1:
The first and second affinity reagents are pre-bound to their respective target analytes or epitopes before the labelling moiety reaction occurs. This preliminary specific binding ensures that only markers with both affinity reagents correctly bound to their targets will form the complete labelling structure, eliminating false positives from cross-reactivity while maintaining high multiplex throughput
Solution Approach 2:
The polymeric backbone with labelling moiety parts acts as an intermediary that bridges the first and second affinity reagents. The labelling moieties only form when both affinity reagents are correctly bound, serving as a mediator that verifies dual-specificity and prevents false-positive signals from single-affinity reagent binding events
3Measurement precision
If markers with bright labels are used to detect small quantities of analytes, then the detection sensitivity is high, but the reliability and specificity are reduced
Solution Approach 1:
The affinity reagents perform preliminary specific binding to the target analytes before the labelling reaction occurs. This ensures that even low-abundance analytes are specifically captured with high reliability, and only then are the bright labelling moieties activated, maintaining both sensitivity and reliability simultaneously
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
Enables precise determination of target analyte proximity and location with reduced false positives, suitable for multiplex assays and diagnostic tests, improving assay specificity and reliability.
Implementation Method 1
The first labelling moiety parts and the second labelling moiety parts are configured to bind to the respective other one to form labelling moieties
Implementation Method 2
The first affinity reagent and the second affinity reagent are each configured to bind specifically to one of the target analytes of the biological sample
Data Source
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AI summary
In a first aspect a marker (100, 404, 500) for analysing a biological sample with a plurality of target analytes (107, 113, 406, 506) is provided. The marker (100, 404, 500) comprises a first marker part (102) comprising a first affinity reagent (106, 400, 502) and a first label, the first label comprising a first polymeric backbone (108) attached to the first affinity reagent (106, 400, 502) and a plurality of first labelling moiety parts (110) attached to the first polymeric backbone (108), and a second marker part (104) comprising a second affinity reagent (112, 402, 504) and a second label, the second label comprising a second polymeric backbone (114) attached to the second affinity reagent (112, 402, 504) and a plurality of second labelling moiety parts (116) attached to the second polymeric backbone (114). The first labelling moiety parts (110) and the second labelling moiety parts (116) are configured to bind to the respective other one (110, 116) to form labelling moieties (118). The first affinity reagent and the second affinity reagent are each configured to bind specifically to one of the target analytes of the biological sample. In a further aspect, a method for analysing a biological sample is provided.