Cross-Linked Nucleic Acid Affinity Reagents for Stable Binding
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Solution Overview
Problem
Nucleic acid based affinity reagents face stability issues due to dynamic 3D structure changes, leading to partial unfolding or misfolding, which complicates their use and storage.
Innovation Solution
A nucleic acid backbone with reaction moieties configured to form cross-links, maintaining a stable complex structure through covalent bonds, such as CuAAC or SpAAC, to enhance stability and specificity in binding to target analytes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If nucleic acid based affinity reagents are used, then cost-effectiveness and ease of manufacture are improved, but structural stability and reliability deteriorate due to dynamic 3D structure changes
Solution Approach 1:
The patent combines nucleic acid backbone with cross-linking moieties to create a composite structure that maintains the cost-effectiveness and ease of manufacture of nucleic acid-based reagents while adding structural stability through cross-linking. The cross-linked nucleic acid affinity reagent integrates the beneficial properties of nucleic acids with enhanced structural rigidity.
Solution Approach 2:
The patent modifies the physical-chemical parameters of the nucleic acid structure by introducing cross-linking bonds that change the conformational flexibility and thermal stability parameters. This allows the reagent to maintain structural integrity across a broader range of temperatures and conditions while retaining the underlying nucleic acid properties.
2Measurement precision
If nucleic acid based affinity reagents are used, then affinity and specificity are improved, but ease of operation deteriorates due to refolding requirements
Solution Approach 1:
The cross-linking moieties are incorporated into the nucleic acid structure in advance during synthesis, creating pre-configured structures that require no post-synthesis refolding steps. The cross-links are formed beforehand to stabilize the active conformation, eliminating the need for operators to perform refolding procedures before use.
Solution Approach 2:
The cross-linked structure provides self-stabilization, maintaining its active conformation without requiring external intervention or refolding steps. The structure serves itself by maintaining structural integrity through the embedded cross-linking moieties, eliminating manual refolding operations.
3Ease of manufacture
If nucleic acid based affinity reagents are used, then cost-effectiveness is improved, but durability deteriorates due to partial unfolding during storage
Solution Approach 1:
The patent creates a composite nucleic acid structure with integrated cross-linking moieties that provide enhanced storage stability while maintaining the cost-effectiveness of nucleic acid-based synthesis. The cross-links act as structural reinforcement that prevents degradation during long-term storage.
Solution Approach 2:
The cross-linking moieties provide preemptive structural protection against unfolding and degradation during storage. By incorporating these stabilizing elements beforehand, the reagent is cushioned against environmental stresses and conformational changes that would otherwise occur during storage, extending its operational lifespan.
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 cross-linked nucleic acid backbone provides a robust and stable affinity reagent with high specificity and resistance to unfolding, enabling efficient analysis of biological samples.
Implementation Method 1
The nucleic acid backbone includes at least one pair of reaction moieties for cross-linking the nucleic acid backbone, such as CuAAC or SpAAC
Implementation Method 2
The nucleic acid backbone includes at least one pair of reaction moieties for cross-linking the nucleic acid backbone, such as CuAAC or SpAAC
Implementation Method 3
The cross-linked nucleic acid backbone provides a robust and stable affinity reagent with high specificity and resistance to unfolding
Data Source
AI summary
An affinity reagent for analysing a biological sample includes a nucleic acid backbone. The nucleic acid backbone is configured to specifically bind to a target analyte by a complex structure of the nucleic acid backbone. The nucleic acid backbone includes at least one pair of reaction moieties for cross-linking the nucleic acid backbone.