Electrochemically-Cleavable Linkers for Aqueous Enzymatic Synthesis
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Solution Overview
Problem
Existing cleavable linker molecules require electrochemical potentials above the redox potential of solvents for cleavage, making them unsuitable for use in certain solvent environments, particularly aqueous solutions where the electrochemical window is narrow and electrolysis of water occurs at relatively low potentials.
Innovation Solution
Development of electrochemically-cleavable linkers with cleavable groups such as methoxybenzyl alcohol, ester, propargyl thioether, trichloroethyl ether, pyrrolidinone-type safety-catch motif, paramethoxy aniline, or ketal, which can cleave at electrochemical potentials below the redox potential of the solvent, allowing for their use in a variety of solvent environments, including aqueous solutions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If existing cleavable linker molecules are used, then they can cleave linkers effectively, but they require electrochemical potentials above the redox potential of solvents which limits their use in certain solvent environments
Solution Approach 1:
The patent modifies the electrochemical properties of linker molecules by introducing specific functional groups (methoxybenzyl alcohol, ester, propargyl thioether, trichloroethyl ether, pyrrolidinone-type safety-catch motif, paramethoxy aniline, or ketal) that lower the cleavage potential below the redox potential of aqueous solvents, enabling effective operation in previously incompatible environments
2Reliability
If electrochemical potentials above the redox potential of solvents are applied, then linker cleavage can be achieved, but electrolysis of water occurs in aqueous solutions at relatively low potentials
Solution Approach 1:
The patent changes the electrochemical parameters of the linker molecules by incorporating functional groups with lower reduction potentials, allowing cleavage to occur at potentials below the water redox potential (approximately -0.83 V vs. SHE at pH 7), thereby preventing water electrolysis while maintaining effective linker cleavage
Solution Approach 2:
The patent replaces traditional chemical cleavage mechanisms with electrochemical cleavage at controlled potentials, using electron transfer reactions to trigger linker breakdown without requiring harsh chemical conditions that would cause solvent decomposition
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 electrochemically-cleavable linkers can effectively cleave in solvents at lower electrochemical potentials than the redox potential of the solvent, enabling their use in challenging environments like aqueous solutions without causing electrolysis of water, thus expanding their applicability in biotechnology applications.
Implementation Method 1
The linkers include at least one cleavable group that may be a methoxybenzyl alcohol, an ester, a propargyl thioether, a trichloroethyl ether, a pyrrolidinone-type safety-catch motif, a paramethoxy aniline, or a ketal. The cleavable group may also optionally include an extension that may be carbonyl group, an alkane group, or an alkene group.
Implementation Method 2
The electrochemically-cleavable linkers provided in this disclosure are cleaved by addition of electrons to a bond in the linker. The electrons may be generated by activating an electrode in the proximity of the bond in the linker that is to be cleaved.
Data Source
AI summary
This disclosure provides electrochemically-cleavable linkers with cleavage potentials that are less than the redox potential of the solvent in which the linkers are used. In some applications, the solvent may be water or an aqueous buffer solution. The linkers may be used to link a nucleotide to a bound group. The linkers include a cleavable group which may be one of a methoxybenzyl alcohol, an ester, a propargyl thioether, or a trichloroethyl ether. The linkers may be cleaved in solvent by generating an electrode potential that is less than the redox potential of the solvent. In some implementations, an electrode array may be used to generate localized electrode potentials which selectively cleave linkers bound to the activated electrode. Uses for the linkers include attachment of blocking groups to nucleotides in enzymatic oligonucleotide synthesis.