Modified Oligonucleotides with Multiple Thiols for Stable Biosensor Immobilization
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Solution Overview
Problem
Current biosensors for detecting nucleic acids face challenges in stability and specificity due to the weakness of Au—S bonds and limited flexibility in hybridization conditions, especially during washing steps, which affects the sensitivity and specificity of nucleic acid detection.
Innovation Solution
Development of modified oligonucleotides with multiple thiol functions that can be efficiently immobilized on gold surfaces or surfaces with carbon-carbon double bonds, triple bonds, or haloacetamide functions, enhancing the stability and specificity of nucleic acid detection regardless of transduction methods used.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional oligonucleotides are used with single thiol function for immobilization on gold surface, then the biosensor can be manufactured simply, but the stability and reliability of immobilization is insufficient due to weak Au-S bonds
Solution Approach 1:
The oligonucleotide is divided into functional segments: a recognition sequence for target binding and multiple thiol functions for immobilization. This segmentation allows the thiol groups to be positioned at specific locations (e.g., 5' end or interspersed) to create multiple Au-S bonds without interfering with the hybridization function of the recognition sequence.
Solution Approach 2:
The invention creates a composite structure by combining oligonucleotide molecules with multiple thiol groups, forming a multi-component system that includes the oligonucleotide backbone, thiol functional groups, and gold surface. This composite approach enables simultaneous achievement of stable immobilization through multiple Au-S bonds and maintained biological activity for target detection.
2Reliability
If multiple thiol functions are introduced into oligonucleotides to strengthen immobilization, then the stability improves, but the manufacturing process becomes more complex
Solution Approach 1:
The invention changes the chemical parameters of the oligonucleotide by introducing thiol-modified nucleotides or thiol groups at specific positions during synthesis. This parameter modification allows standard solid-phase synthesis methods to be used while incorporating multiple thiol functions, thereby improving immobilization stability without fundamentally altering the manufacturing process.
3Measurement precision
If conventional biosensors are used with limited flexibility in hybridization conditions, then the device structure remains simple, but the measurement precision and detection sensitivity are reduced
Solution Approach 1:
The invention introduces dynamic flexibility into the oligonucleotide structure through multiple thiol functions that can adapt to different immobilization configurations and hybridization conditions. This dynamic capability allows the biosensor to maintain stable immobilization while accommodating variations in hybridization temperature, salt concentration, and washing stringency, thereby improving detection sensitivity without excessive structural complexity.
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 modified oligonucleotides provide improved stability and specificity for detecting target nucleic acid sequences, enabling more sensitive and flexible detection methods for pathogenic organisms or genes, which are economical and easier to automate.
Implementation Method 1
modified oligonucleotide having two or more thiol functions, which can be immobilized on a gold surface or on a grafted surface, in particular a surface comprising at least one carbon-carbon double bond or carbon-carbon triple bond or haloacetamide functions
Implementation Method 2
detecting hybridization between a modified oligonucleotide and a target nucleic acid amplified from the biological sample
Implementation Method 3
The molecular recognition element, called 'probe', is generally immobilized on the surface of the transducer and displays high specificity and sensitivity for a 'target' nucleic acid molecule
Data Source
AI summary
The present invention relates to a modified oligonucleotide having two or more thiol functions, which can be immobilized on a gold surface or on a grafted surface, in particular a surface comprising at least one carbon-carbon double bond or carbon-carbon triple bond or haloacetamide functions, preferably maleimide or acrylamide functions. The invention also relates to a method for detecting a nucleic acid in a biological sample comprising a step of detecting hybridization between a modified oligonucleotide and a target nucleic acid amplified from the biological sample. The invention relates more particularly to a method for detecting, genotyping or sequencing a pathogenic organism, preferably a virus.