Dual-Part Nucleic Acid Labels for Specific Proximity Detection
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Solution Overview
Problem
Existing biological sample analysis methods face challenges with cross-reactivity of affinity reagents, leading to false-positive results and reduced specificity, especially in high-plex assays, which are exacerbated by high background autofluorescence in tissue samples.
Innovation Solution
A label comprising a first and second nucleic acid backbone with complementary sequences and optically detectable moieties, along with a guest-host system to control duplex formation, allowing proximity detection with high specificity and minimal background interference.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional affinity reagents with detectable labels are used for biological sample analysis, then detection capability is achieved, but cross-reactivity occurs leading to false-positive results and reduced specificity
Solution Approach 1:
The label is divided into two separate label parts (first label part and second label part) that only come together to form a complete detectable signal when both affinity reagents bind to their respective target analytes in close proximity. This segmentation prevents false-positive signals from single affinity reagent binding events, thereby resolving the contradiction between detection capability and false-positive rate.
Solution Approach 2:
The first and second label parts act as intermediaries that require both affinity reagents to be present and in close proximity to form a complete detectable complex. This intermediary mechanism ensures that detection specificity is maintained while preventing false-positive results from non-specific binding of individual affinity reagents.
2Adaptability or versatility
If multiple markers are used for high-plex assays, then multiplexing capability is achieved, but handling complexity increases
Solution Approach 1:
The dual-label architecture provides a universal platform that can be applied across multiple affinity reagent pairs and target analytes. The same first and second label parts can work with different affinity reagents, enabling high-plex assays without proportionally increasing handling complexity, as the detection mechanism remains consistent across multiple targets.
3Measurement precision
If conventional labels are used in tissue samples, then detection is achieved, but background autofluorescence interferes with signal detection
Solution Approach 1:
By segmenting the label into two parts that only form a complete signal when both affinity reagents are bound in close proximity, the invention minimizes background signal from unbound or singly-bound affinity reagents. This segmentation strategy effectively reduces the impact of tissue autofluorescence by ensuring that only specific proximal binding events generate detectable signals above the background noise.
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 accurate detection of analyte proximities with reduced cross-reactivity and background noise, suitable for high-plex assays and tissue samples, using FRET and dequenching mechanisms for robust readout.
Implementation Method 1
The first nucleic acid backbone and the second nucleic acid backbone are each configured to hybridise to the respective other one
Implementation Method 2
The label comprises at least one guest molecule configured to form a complex with a host molecule
Implementation Method 3
using FRET and dequenching mechanisms for robust readout
Implementation Method 4
using FRET and dequenching mechanisms for robust readout
Data Source
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AI summary
In a first aspect a label for analysing a biological sample is provided. The label comprises a first label part comprising a first nucleic acid backbone (100, 206), and a second label part comprising a second nucleic acid backbone (102, 210). The first nucleic acid backbone (100, 206) and the second nucleic acid backbone (102, 210) are each configured to hybridise to the respective other one. The label further comprises at least one first labelling moiety (208) and at least one second labelling moiety (212). The label further comprises at least one guest molecule (104) configured to form a complex with a host molecule (106). In further aspects, a corresponding marker (200, 500, 600) and a method for analysing a biological sample are provided.