Polyribonucleotide Purification via Oligonucleotide Hybridization
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Solution Overview
Problem
Current methods for separating and purifying polyribonucleotides are inefficient, particularly in distinguishing between polyribonucleotides with target regions and those without, especially when the target regions are not located at the 3′ or 5′ termini or lack polyA sequences, leading to impurities and incomplete products in therapeutic and engineering applications.
Innovation Solution
A method involving the use of oligonucleotides that hybridize to target regions within polyribonucleotides, allowing for selective separation by differential binding affinities, using conjugated capture agents and particles to immobilize and purify polyribonucleotides with specific target regions, even when these regions are internally located or absent in circular forms.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional separation methods are used, then the process is simple, but the purification efficiency is low and cannot distinguish polyribonucleotides with internal target regions or without polyA sequences
Solution Approach 1:
The patent introduces an oligonucleotide as an intermediary that hybridizes to the target region of polyribonucleotides. This oligonucleotide acts as a mediator between the polyribonucleotide and the capture agent, enabling specific recognition and separation of polyribonucleotides with internal target regions or without polyA sequences, thereby significantly improving purification efficiency
Solution Approach 2:
The patent extracts the target region recognition function from conventional separation methods by using a specifically designed oligonucleotide that hybridizes only to the target region. This allows the method to selectively extract and separate polyribonucleotides containing the target region from the mixture, achieving high purification efficiency without relying on polyA sequences or terminal locations
2Manufacturing precision
If oligonucleotide hybridization is used to separate polyribonucleotides with internal target regions, then purification precision is improved, but the process time increases
Solution Approach 1:
The patent optimizes hybridization parameters including temperature, salt concentration, and oligonucleotide concentration to achieve rapid and specific hybridization. By carefully controlling these parameters, the method achieves high purification precision while minimizing the time required for the hybridization and separation process
3Reliability
If capture agents are used to immobilize polyribonucleotides, then separation effectiveness is improved, but the device complexity increases
Solution Approach 1:
The patent uses capture agents as intermediaries that bind to the oligonucleotide-polyribonucleotide complex. This intermediary approach allows effective immobilization and separation of polyribonucleotides while maintaining a relatively simple system architecture, as the capture agent serves as a bridge between the hybridized complex and the solid support
Solution Approach 2:
The patent employs capture agents with universal binding properties that can work with various oligonucleotide designs and polyribonucleotide targets. This multi-functionality reduces the need for specialized components for each specific application, thereby maintaining system simplicity while achieving high separation effectiveness
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 effectively enriches polyribonucleotides with target regions by up to 99% relative to the sample, reducing impurities and improving the purity and usability of polyribonucleotides for therapeutic and engineering purposes.
Implementation Method 1
contacting the sample with an oligonucleotide that hybridizes to the target region
Data Source
AI summary
The present disclosure relates to compositions and methods for separating and/or purifying polyribonucleotides. The polyribonucleotide may be separated from a mixture of polyribonucleotides with an oligonucleotide that hybridizes to a target region of the polyribonucleotide and be available for use as a therapeutic agent.