Nanofiltration Purification of Oligonucleotide Synthons
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Solution Overview
Problem
Current methods for purifying synthons used in oligonucleotide synthesis, such as phosphoramidites and H-phosphonates, are inefficient and time-consuming, particularly at large scales, where kilogram quantities are needed for clinical trials, and existing chromatography methods are lengthy and ineffective for large-scale preparation.
Innovation Solution
A nanofiltration process is employed to purify oligonucleotide synthons, specifically subjecting an organic solution containing an oligonucleotide synthon and lower molecular weight impurities to nanofiltration, increasing the ratio of synthon to impurities, using membranes with a molecular weight cut-off at about 400, and employing high pressure and suitable solvents like ethyl acetate to concentrate and purify protected nucleoside phosphoramidites and H-phosphonates.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If chromatography methods are used to purify phosphoramidites and H-phosphonates, then some level of purification is achieved, but the process becomes lengthy and time-consuming, especially at large scales
Solution Approach 1:
The patent replaces the mechanical chromatography system with a nanofiltration system that uses a semi-permeable membrane to separate synthons from impurities based on molecular size. This substitution eliminates the lengthy chromatography process while achieving comparable or superior purification efficiency, directly addressing the time loss issue.
Solution Approach 2:
The patent changes the purification parameter from chromatographic separation (based on chemical affinity) to nanofiltration separation (based on molecular weight cutoff). By using a membrane with a specific molecular weight cutoff, the process achieves rapid separation of synthons from lower molecular weight impurities, dramatically reducing purification time.
2Manufacturing precision
If chromatography is used for purification, then purify compounds can be obtained, but the process is ineffective for large-scale preparation where kilogram quantities are needed
Solution Approach 1:
The patent replaces chromatography with nanofiltration, which is inherently scalable. The membrane-based separation process can handle large volumes and kilogram quantities of synthons without the capacity limitations that plague chromatographic methods, enabling effective large-scale preparation.
Solution Approach 2:
The patent changes from a labor-intensive, capacity-limited chromatographic process to a continuous nanofiltration process that can be scaled up. The molecular weight cutoff parameter allows for straightforward scaling to handle kilogram quantities while maintaining purification effectiveness.
3Manufacturing precision
If existing purification methods are used for H-phosphonates and phosphoramidites, then some purification is achieved, but the methods suffer from similar disadvantages to phosphoramidites including time consumption and inefficiency at large scales
Solution Approach 1:
The patent develops a universal nanofiltration process that can purify multiple types of synthons (H-phosphonates, phosphoramidites, and other oligonucleotide building blocks) using the same membrane-based approach. This multi-functional method eliminates the need for different purification protocols for different synthon types, reducing overall time consumption.
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 process significantly increases the purity and concentration of oligonucleotide synthons, reducing the volume of impurities and efficiently preparing high-purity synthons for large-scale therapeutic applications, such as CMV treatment, by selectively allowing lower molecular weight impurities to pass through the membrane while retaining the synthons, thus addressing the inefficiencies of existing purification methods.
Implementation Method 1
subjecting an organic solution comprising an oligonucleotide synthon and lower molecular weight impurities to nanofiltration whereby the ratio of an oligonucleotide synthon to lower molecular weight impurities in the solution is increased after the nanofiltration
Implementation Method 2
using membranes with a molecular weight cut-off at about 400, and employing high pressure and suitable solvents like ethyl acetate to concentrate and purify protected nucleoside phosphoramidites and H-phosphonates
Implementation Method 3
employing high pressure and suitable solvents like ethyl acetate to concentrate and purify protected nucleoside phosphoramidites and H-phosphonates
Data Source
AI summary
A process for the purification of an oligonucleotide synthon is provided. The process comprises subjecting an organic solution comprising an oligonucleotide synthon and lower molecular weight impurities to nanofiltration whereby the ratio of an oligonucleotide synthon to lower molecular weight impurities in the solution is increased after the nanofiltration. Preferably, the oligonucleotide synthon is a nucleoside phosphoramidite or nucleoside H-phosphonate. The nanofiltration membrane is preferably a polyimide membrane having a molecular weight cut off of 400.