Nanopore Sequencing Electrolyte Stability via Cation Complexing

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Solution Overview

Problem

Nanopore sequencing techniques face challenges due to partial consumption of electrolyte redox reagents, leading to concentration depletion and current drift, which affects the ability to accurately distinguish nucleotide bases.

Innovation Solution

Mitigating the diffusion of electrochemically passive species through the nanopore by using cation complexing agents, such as crown ethers, to exclusively carry charge with anions, reducing reagent depletion and maintaining a stable ionic balance.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If nanopore sequencing is performed using conventional electrolyte solutions, then the sequencing process can proceed, but electrolyte redox reagents are partially consumed leading to concentration depletion and current drift

Engineering Contradiction:
Improvesequencing throughputVSAvoidcurrent reading accuracy
Core Design Contradiction:
ProductivityVSReliability

Solution Approach 1:

The patent changes the chemical composition parameters of the electrolyte solution by introducing cation complexing agents (such as crown ethers) that selectively bind cations. This parameter change prevents cation diffusion through the nanopore, thereby preventing concentration depletion of redox reagents and eliminating current drift, while maintaining sequencing productivity

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent introduces cation complexing agents as intermediary substances that mediate between the electrolyte ions and the nanopore. These agents form complexes with cations, preventing their direct diffusion through the nanopore while allowing anions to carry charge, thus maintaining ionic balance and preventing reagent depletion

Inventive Principle:
Principle #24Intermediary (Mediator)

2Reliability

If cation complexing agents are used to prevent cation diffusion, then reagent depletion is reduced to less than 10%, but the device complexity increases

Engineering Contradiction:
Improveelectrolyte stabilityVSAvoidelectrolyte composition complexity
Core Design Contradiction:
ReliabilityVSDevice complexity

Solution Approach 1:

The patent modifies the electrolyte composition by adding specific cation complexing agents at controlled concentrations. This parameter change achieves reliable prevention of cation diffusion and reagent depletion while maintaining manageable device complexity through selective use of well-characterized chemical agents

Inventive Principle:
Principle #35Parameter changes

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 reduces reagent depletion to less than 10%, maintaining accurate current readings and extending the lifetime of electrolytes and electrodes, thereby improving the reliability of nucleotide base identification in nanopore sequencing.

Implementation Method 1

Mitigating the diffusion of electrochemically passive species through the nanopore by using cation complexing agents, such as crown ethers, to exclusively carry charge with anions

Methodology Applied
Scientific EffectComplexation:

Implementation Method 2

A polynucleotide is driven through the nanopore, and as the polynucleotide passes through the nanopore, it disrupts the electrical current through the nanopore

Methodology Applied
Scientific EffectIonic conduction: Conduction (electrical)

Implementation Method 3

One electrolyte redox reagent may be partially consumed on a trans well (or chamber) electrode in order to support Faradaic current through the system

Methodology Applied
Scientific EffectRedox reactions: Redox Reactions

Data Source

PatentEP3642619B1Nanopore sequencing method
Publication Date: 2024.07.31 ILLUMINA INC
  • EP3642619B1 patent drawingFigure 1~2
  • EP3642619B1 patent drawingFigure 3~4
  • EP3642619B1 patent drawingFigure 5~6A

AI summary

Example nanopore sequencers include a cis well, a trans well, and a nanopore fluidically connecting the cis and trans wells. In one example sequencer, a modified electrolyte (including an electrolyte and a cation complexing agent) is present in the cis well, or the trans well, or in the cis and the trans wells. In another example sequencer, a gel state polyelectrolyte is present in the cis well, or the trans well, or in the cis and the trans wells.