Biological Sample Markers with Covalent Linkers for Robust Assembly
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Solution Overview
Problem
Existing markers for biological samples, such as those used in fluorescent microscopy, are complex to produce due to individual assembly of parts, leading to inefficiencies and reduced robustness, with reversible attachments resulting in shorter shelf life.
Innovation Solution
A marker comprising an affinity reagent, a linker oligonucleotide, and a labelling moiety, where the linker oligonucleotide is covalently attached to both, enabling efficient and robust assembly without hybridization, using methods like click chemistry and ligase-mediated ligation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If individual parts of the marker are assembled through hybridization, then the marker can be produced modularly, but the attachment becomes reversible and less robust
Solution Approach 1:
The patent replaces the mechanical hybridization-based attachment system with a covalent bonding system. Specifically, it uses click chemistry (copper-catalyzed azide-alkyne cycloaddition) to form irreversible covalent bonds between the affinity reagent and the oligonucleotide linker, and between the oligonucleotide and the label. This substitution of bonding mechanism maintains modular assembly capability while dramatically improving attachment robustness and marker stability.
2Adaptability or versatility
If individual parts are assembled step-by-step, then the marker can be constructed with specific components, but the production process becomes complex and time-consuming
Solution Approach 1:
The patent applies preliminary action by pre-functionalizing the affinity reagent with an azide group and the oligonucleotide linker with an alkyne group before assembly. This pre-preparation of reactive groups enables a single-step click chemistry conjugation reaction, simplifying the overall process while maintaining the ability to selectively choose different affinity reagents, linkers, and labels for specific applications.
3Ease of manufacture
If traditional marker assembly methods are used, then individual components can be optimized separately, but reaction losses and purification losses increase
Solution Approach 1:
The patent replaces traditional step-by-step chemical conjugation methods with click chemistry, which offers high reaction efficiency and specificity. The copper-catalyzed azide-alkyne cycloaddition proceeds with near-quantitative yield and minimal side reactions, eliminating the need for extensive purification steps. This substitution dramatically reduces both reaction losses and purification losses while maintaining the ability to optimize individual components separately.
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
The solution allows for efficient, robust, and bright markers to be generated with reduced losses, facilitating easy assembly and analysis of biological samples without the need for separation or purification steps.
Implementation Method 1
The at least one linker oligonucleotide is attached to the affinity reagent, and the at least one labelling moiety is attached to the at least one linker oligonucleotide
Implementation Method 2
using methods like click chemistry and ligase-mediated ligation
Data Source
AI summary
A marker for analysing a biological sample includes an affinity reagent, at least one labelling moiety, and at least one linker oligonucleotide. The at least one linker oligonucleotide is attached to the affinity reagent, and the at least one labelling moiety is attached to the at least one linker oligonucleotide.