Cleavable Linker Polymer Synthesis for Homogeneous Product Pools
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current DNA synthesis methods, particularly solid-phase synthesis, face challenges with heterogeneous product pools due to retention of unreacted nascent DNA strands, leading to undesirable lengths and purification difficulties.
Innovation Solution
The alternating-phase polymer and nucleic acid synthesis method involves immobilizing monomers or nucleotides to a support via a cleavable linker, allowing for controlled coupling and removal of uncoupled species, and subsequent cleavage of the extended polymer or nucleic acid from the support, thereby avoiding immobilization of unreacted strands and minimizing contamination.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If solid-phase synthesis is used to synthesize DNA, then purification efficiency is improved and product loss is minimized, but heterogeneous product pools are formed due to retention of unreacted nascent DNA strands
Solution Approach 1:
The patent extracts the problematic feature (permanent immobilization) from the solid-phase synthesis method. By making immobilization reversible through cleavable linkers, the method removes unreacted strands from the support after each coupling cycle, preventing them from contaminating the final product pool while maintaining the purification efficiency of solid-phase synthesis.
Solution Approach 2:
The patent changes the chemical state of the linker from permanent to cleavable. This parameter change allows the support-monomer conjugate to be formed initially for efficient purification, then subsequently cleaved to release the synthesized product free from heterogeneous contaminants, thus resolving the contradiction between purification efficiency and product homogeneity.
2Ease of manufacture
If monomers are permanently immobilized to support in solid-phase synthesis, then purification is simplified, but unreacted nascent strands cannot be removed leading to product heterogeneity
Solution Approach 1:
The patent introduces dynamics to the previously static immobilization system. The linker transitions from a permanent, fixed state to a dynamic, conditionally cleavable state. This allows the system to adapt: remaining immobilized during synthesis for easy purification, then becoming mobile/cleaved for product release, thereby achieving both purification simplicity and product purity.
Solution Approach 2:
The patent implements periodic action through the cyclical formation and cleavage of the support-monomer conjugate. The linker is formed during synthesis cycles to enable purification, then periodically cleaved to release the final product. This periodic switching between immobilized and free states resolves the contradiction between ease of manufacture and product purity.
3Productivity
If large excess of reagent is used to increase N to N+1 conversion rates, then coupling efficiency is improved, but product homogeneity deteriorates due to retention of unreacted strands
Solution Approach 1:
The patent converts the harmful effect of unreacted strand retention into a beneficial feature. Instead of viewing unreacted strands as contamination to be avoided, the method allows them to remain on the support temporarily, then uses the cleavable linker to selectively remove them in a subsequent step. This transforms the problem of unreacted strand retention into an opportunity for enhanced product homogeneity while maintaining high coupling efficiency.
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 approach eliminates contaminants like X−1 species, ensures controlled exposure to reagents, and avoids polymerase steric gate clashes, resulting in a more efficient and error-corrected synthesis process with homogeneous product pools.
Implementation Method 1
movement of the initiator nucleic acid sequence between areas of immobilized dNTPs/NTPs can be achieved by droplet actuation by electrowetting on dielectric (EWOD)
Data Source
AI summary
The invention relates to methods of synthesizing polymers, biopolymers, and nucleic acids, to immobilised dNTP/NTPs and kits comprising said immobilised dNTP/NTPs for use in said methods of nucleic acid synthesis.