Freeze-Thaw Protective Polymer for Nucleic Acid Amplification
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Solution Overview
Problem
Existing nucleic acid amplification compositions require high concentrations of additives like sugars, glycerin, and DMSO for freeze-thaw protection, which can affect protein stability and necessitate removal operations, and DMSO poses toxicity risks.
Innovation Solution
A polymer containing constitutional units derived from specific monomers, such as 2-(meth)acryloyloxyethyl phosphorylcholine, is used as a freeze-thawing protective agent for nucleic acid amplification compositions, offering protection at lower concentrations without the need for post-thaw removal.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If sugars, glycerin, or albumin are added as freeze-thawing protective agents, then deactivation due to freeze-thawing is reduced, but extremely high concentrations are required and removal operations are needed
Solution Approach 1:
The patent changes the chemical nature of the protective agent from small molecules (sugars, glycerin) to a specific polymer structure with phosphorylcholine groups. This parameter change allows achieving protective effects at much lower concentrations (0.01-10% w/v) compared to conventional agents that require extremely high concentrations
Solution Approach 2:
The protective agent is a copolymer combining phosphorylcholine-containing monomer units with other compatible monomer units. This composite structure provides both the freeze-thaw protection capability and compatibility with nucleic acid amplification reagents, eliminating the need for removal operations
2Reliability
If DMSO is used as a freeze-thawing protective agent, then deactivation is reduced, but toxicity to proteins and cell metabolism occurs
Solution Approach 1:
The patent replaces toxic DMSO with a biocompatible polymer that can remain in the composition without causing harm. The polymer acts as a disposable protective element that maintains reagent stability through freeze-thaw cycles without introducing toxicity to subsequent biological applications
Solution Approach 2:
The phosphorylcholine-containing polymer serves as an intermediary substance that provides freeze-thaw protection through steric stabilization and hydration shell formation, rather than through the toxic mechanism of DMSO. This intermediary approach achieves protection without the harmful side effects
3Reliability
If sugars, glycerin, or albumin are added as freeze-thawing protective agents, then deactivation is reduced, but the properties of target substances may be affected
Solution Approach 1:
The patent introduces localized phosphorylcholine groups at specific positions in the polymer structure. These localized functional groups provide freeze-thaw protection through electrostatic repulsion and hydration, while the rest of the polymer structure remains inert and compatible with nucleic acid amplification reactions, thus preserving target substance properties
Data Source
AI summary
The present invention provides a freeze-thawing protective agent for a composition for nucleic acid amplification, which is a polymer containing constitutional units derived from a monomer represented by the following formula (1) (the definitions of symbols in the formula are as described in the specification).