Aptamer Selection via Inverted Solid-Phase Hybridization
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Solution Overview
Problem
Current methods for selecting and isolating aptamers specific to particular targets are inefficient and time-consuming, lacking rapid and effective strategies for identifying and isolating aptamers with high affinity and specificity.
Innovation Solution
A novel method involving structure-switching aptamers, where a library of oligonucleotides with random and antisense binding domains is hybridized, immobilized, and then incubated with a target to selectively release aptamers that bind, allowing for amplification and sequencing of specific aptamer sequences, enabling the selection of aptamers capable of binding to ATP and GTP.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If the traditional SELEX process is used to select aptamers, then aptamers with high affinity and specificity can be obtained, but the selection process is time-consuming and inefficient
Solution Approach 1:
Instead of immobilizing the target and allowing library molecules to bind (traditional SELEX), the patent inverts the approach by immobilizing the library molecules on a solid support and allowing target molecules to bind to them in solution. This inversion simplifies the selection process and reduces time while maintaining aptamer specificity.
Solution Approach 2:
The patent replaces the complex mechanical column flushing and elution system of traditional SELEX with a simpler solid-phase binding system. Library molecules are covalently attached to solid support, and target binding is detected directly without requiring column manipulation, significantly reducing process time.
2Strength
If multiple SELEX cycles are repeated to achieve higher affinity, then aptamer binding strength increases, but the overall process becomes more complex and time-consuming
Solution Approach 1:
The patent enables continuous selection by maintaining library molecules in a fixed state on solid support throughout multiple selection cycles. The same solid-phase library can be repeatedly exposed to different target molecules without re-immobilization, allowing continuous affinity improvement without increasing procedural complexity.
3Adaptability or versatility
If a large library of random DNA molecules is used to ensure target coverage, then the probability of finding specific aptamers increases, but the volume of material to be processed increases
Solution Approach 1:
The patent segments the library into individual molecules covalently attached to solid support surfaces, creating a distributed array of potential aptamers. This segmentation allows each library member to be independently accessible to targets in solution, maintaining diversity while reducing the bulk volume that needs to be processed compared to traditional solution-phase libraries.
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
This method enables the rapid and efficient selection of aptamers with high specificity and affinity, allowing for the conversion of selected aptamers into signaling aptamers for biological detection, enhancing their application in medical diagnosis and biosensing.
Implementation Method 1
hybridizing the library oligonucleotide to a biotinylated antisense oligonucleotide to form a duplex molecule
Implementation Method 2
interacting the duplex molecule with avidin coated beads to immobilize the duplex molecule on the beads
Implementation Method 3
incubating the duplex structure in the presence of the target; and collecting library oligonucleotides that dissociate from the duplex structure and bind to the target
Data Source
AI summary
The present invention provides a method for the selection of aptamer sequences and aptamers identified using the method. The method comprises immobilizing a library oligonucleotide by forming a duplex with an antisense oligonucleotide, reating the duplex molecule with a target and collecting oligonucleotide molecules which dissociate from the duplex structure.