Bi-functional pH-switchable compound for nucleic acid isolation
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Solution Overview
Problem
Current methods for isolating nucleic acids using pH-dependent ion exchange materials are inefficient in binding and releasing nucleic acids, as they lack effective bi-functional compounds that are positively charged at a first pH and negatively charged at a second pH.
Innovation Solution
A bi-functional compound represented by Formula I, which is positively charged at a first pH and negatively charged at a second pH, is immobilized on a solid support, derived from a sequential reaction product of a substrate, a surface treatment agent, a dianhydride or polyanhydride, and a nitrogen-containing compound, allowing for efficient binding and release of nucleic acids at different pH levels.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If pH-dependent ion exchange materials with single ionizable groups are used, then nucleic acid binding is achieved, but release efficiency is insufficient
Solution Approach 1:
The patent applies parameter changes by using a bi-functional compound where the charge state changes with pH. At low pH, the compound is positively charged for binding; at high pH, it becomes negatively charged for release. This dynamic parameter change resolves the contradiction between binding and release efficiency.
Solution Approach 2:
The patent uses a composite material structure with two different ionizable groups (amino group and carboxyl group) in the same compound. The amino group provides positive charge at low pH for binding, while the carboxyl group provides negative charge at high pH for release, creating a material that performs both functions.
2Ease of manufacture
If common buffer compounds are used as ion exchange materials, then nucleic acid isolation is possible, but binding and release efficiency is insufficient
Solution Approach 1:
The patent modifies common buffer compounds by introducing dual ionizable groups that change charge state with pH. This parameter change enables the material to both bind and release nucleic acids efficiently, while still using commercially available starting materials.
3Reliability
If silica-coated magnetic particles with ion exchange ligands are used, then nucleic acid binding is achieved, but efficient release at higher pH is not sufficient
Solution Approach 1:
The patent applies parameter changes by designing a compound where pH controls the charge state. At the higher pH used for release, the carboxyl group becomes deprotonated providing negative charge that repels the negatively charged nucleic acid, enabling efficient release.
Solution Approach 2:
The patent combines silica-coated magnetic particles with bi-functional compounds containing both amino and carboxyl groups. This composite structure provides both the magnetic separation capability and the pH-dependent charge reversal for efficient binding and release.
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 bi-functional compound effectively binds nucleic acids at a lower pH and releases them at a higher pH, enhancing the efficiency of nucleic acid isolation with high binding and elution efficiencies, as demonstrated in various examples using silicon substrates and magnetic beads.
Implementation Method 1
a bi-functional compound, positively charged at a first pH and negatively charged at a second pH
Implementation Method 2
a bi-functional compound, positively charged at a first pH and negatively charged at a second pH
Implementation Method 3
a material having an ionizable group, such as an acidic functional group or basic functional group
Data Source
AI summary
Provided are a bi-functional compound that is positively charged at a first pH and negatively charged at a second pH, a solid support having the bi-functional compound immobilized thereon, and a method of isolating a nucleic acid, including: binding the bi-functional compound with a nucleic acid at a first pH and isolating the nucleic acid from the bi-functional compound at a second pH.