Electrochemically-Cleavable Linkers Below Water Electrolysis Potential
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Solution Overview
Problem
Existing cleavable linkers are unsuitable for use in aqueous environments due to the high electrochemical potential required for cleavage, which often leads to electrolysis of water, limiting their application in enzymatic nucleotide synthesis and sequencing-by-synthesis techniques.
Innovation Solution
Development of electrochemically-cleavable linkers with cleavage potentials below the redox potential of solvents, allowing for controlled cleavage using electrode potentials less than the solvent's hydrolysis potential, enabling selective cleavage on electrode arrays and facilitating applications in aqueous environments.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If high electrochemical potential is used to cleave existing linkers, then linker cleavage is achieved, but water electrolysis occurs
Solution Approach 1:
The patent modifies the electrochemical parameters of the linker by incorporating redox-active groups (such as quinone, catechol, or disulfide groups) that enable cleavage at lower potentials. These groups undergo reversible redox reactions at potentials below the water electrolysis threshold, generating reactive species that trigger linker cleavage without causing harmful water decomposition
Solution Approach 2:
The patent introduces redox-mediated cleavage mechanisms where a mediator molecule (such as a quinone or disulfide group embedded in the linker) acts as an intermediary. The mediator undergoes redox cycling at low potentials and transfers electrons to the cleavage site, enabling indirect cleavage without applying high potentials that would electrolyze water
2Reliability
If traditional cleavage methods are used in aqueous environments, then cleavage can occur, but the process lacks precision and control
Solution Approach 1:
The patent enables spatially selective linker cleavage by applying electrochemical potential to specific electrode locations. Only linkers in proximity to activated electrodes undergo cleavage, while those in other regions remain intact. This local electrochemical activation provides precise spatial control over where cleavage occurs in the aqueous environment
Solution Approach 2:
The patent employs temporal control of cleavage through periodic or pulsed electrochemical activation. By controlling the timing and duration of potential application, the system can trigger cleavage at specific time points, enabling sequential or synchronized release of multiple linkers with precise temporal resolution
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
Enables efficient and controlled cleavage of linkers in aqueous solvents, allowing for precise enzymatic nucleotide synthesis and sequencing-by-synthesis, overcoming the limitations of traditional cleavage methods that cause water electrolysis.
Implementation Method 1
The linker may be reduced or oxidized, thereby triggering a fragmentation reaction that cleaves the bond between the nucleotide and the bound group
Implementation Method 2
The linker may be reduced or oxidized, thereby triggering a fragmentation reaction that cleaves the bond between the nucleotide and the bound group
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.