Nucleic Acid Purification via Neutral-pH Nucleophile Treatment
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Nucleic acids produced by chemical synthesis often contain a by-product with a mass 34 daltons greater than the target nucleic acid, which has similar molecular weight and physical properties, making it difficult to remove using existing purification methods.
Innovation Solution
Synthesize nucleic acids with an amino group of a base moiety protected by an acyl protecting group, then treat them with a nucleophile under neutral or acidic pH conditions, followed by deprotecting the protecting group to reduce the by-product content.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If known purification methods (RPC, RP HPLC, AEX, PAGE) are used, then general purification is achieved, but the by-product with mass 34 daltons greater than the target nucleic acid cannot be removed due to similar molecular weight and physical properties
Solution Approach 1:
The invention performs a preliminary treatment step before final purification by contacting the synthesized nucleic acid with a nucleophile under neutral or acidic pH conditions. This preliminary action modifies the by-product or target molecule in advance, creating a difference that enables subsequent effective separation and purification, thereby resolving the contradiction between maintaining high purity and dealing with similar molecular weight compounds.
2Ease of manufacture
If the amino group is protected with an acyl protecting group during synthesis, then the target nucleic acid can be produced, but the by-product with 34 daltons mass difference is generated and difficult to remove
Solution Approach 1:
The invention changes the pH parameter from conventional basic conditions to neutral or acidic conditions during the nucleophile treatment step. This parameter change enables selective reaction with the by-product or target molecule, creating a distinguishable difference that facilitates purification. The acyl protecting group strategy is maintained for synthesis feasibility while the pH modification enables subsequent purity improvement.
3Reliability
If strict quality control for impurities is implemented, then safety is ensured, but additional purification steps are required increasing process complexity
Solution Approach 1:
The invention performs a preliminary nucleophile treatment step that simplifies subsequent purification by creating a greater difference between the target nucleic acid and by-products. This preliminary action reduces the complexity of downstream purification processes while maintaining strict quality control, thereby ensuring safety without excessive process 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 method effectively reduces the by-product content to 0.50 or less relative to the target nucleic acid, achieving high purity nucleic acids.
Implementation Method 1
bringing the nucleic acid obtained in step (A) into contact with a nucleophile under a condition of a neutral or acidic pH
Implementation Method 2
deprotecting the protecting group of the nucleic acid obtained in step (B)
Data Source
AI summary
A nucleic acid production method includes: (A) synthesizing a nucleic acid in which an amino group of a base moiety is protected with an acyl protecting group; (B) bringing the nucleic acid obtained in step (A) into contact with a nucleophile under a condition of a neutral or acidic pH; and (C) deprotecting the protecting group for the nucleic acid obtained in step (B). An embodiment of the present invention reduces the content of a by-product (B), which cannot be removed by a known technique, and can produce a nucleic acid with high purity.