Imide Polymer Coating for Stable DNA Attachment in Flow Cells

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Solution Overview

Problem

Current microfluidic flow cell devices face challenges in maintaining stable attachment of DNA fragments during sequencing cycles, leading to limited read lengths due to the instability of DNA-substrate linkages, particularly with amide bond-based attachments.

Innovation Solution

The introduction of a flow cell system with a substrate having a fluidic channel coated with a polymer containing imide functional groups, formed through copolymerization with maleic anhydride, which covalently attaches nucleic acid primer molecules, providing a stable and reactive surface for DNA fragment capture and sequencing.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Ease of manufacture

If amide bond-based attachment is used to attach DNA fragments to the substrate, then the attachment process is simple and widely applicable, but the stability of DNA attachment deteriorates during sequencing cycles leading to limited read lengths

Engineering Contradiction:
Improveattachment process simplicityVSAvoidDNA attachment stability
Core Design Contradiction:
Ease of manufactureVSReliability

Solution Approach 1:

The patent changes the chemical parameter of the linkage bond from amide to imide. The imide bond provides enhanced chemical stability and resistance to hydrolysis compared to amide bonds, directly addressing the reliability issue while maintaining the covalent attachment approach that ensures ease of manufacture

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent employs a composite structure consisting of the substrate, coupling agent, polymer coating with imide functional groups, and DNA fragment. This multi-layer composite system provides both the simplicity of covalent attachment and the enhanced stability of imide bonds, resolving the contradiction between ease of manufacture and attachment stability

Inventive Principle:
Principle #40Composite materials

2Productivity

If multiple rigorous washing steps are performed during sequencing cycles, then the sequencing process can be completed, but DNA molecules are lost over time limiting read length

Engineering Contradiction:
Improvesequencing cycle completionVSAvoidDNA molecule loss
Core Design Contradiction:
ProductivityVSLoss of substance

Solution Approach 1:

The patent applies beforehand cushioning by pre-attaching DNA fragments to the substrate using stable imide bonds before the sequencing cycles begin. This strong initial attachment prevents DNA loss during the subsequent rigorous washing steps, allowing complete sequencing cycles to be performed without significant molecule loss

Inventive Principle:
Principle #11Beforehand cushioning (Prior cushioning)

3Duration of action of moving object

If longer sequencing read lengths are achieved, then more comprehensive DNA analysis is possible, but the attachment stability must be improved to withstand extended cycling conditions

Engineering Contradiction:
Improvesequencing read lengthVSAvoidDNA-substrate linkage stability
Core Design Contradiction:
Duration of action of moving objectVSStability of the object's composition

Solution Approach 1:

The patent changes the chemical composition parameter of the linkage from amide to imide bonds. This chemical parameter change provides the enhanced stability required to maintain DNA attachment integrity over extended sequencing cycles, enabling longer read lengths without compromising linkage stability

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 enhances the stability and reliability of DNA attachment, allowing for longer sequencing read lengths and improved precision in DNA analysis, as demonstrated by the comparison of imide linkage stability against amide linkage in sequencing by synthesis methods.

Implementation Method 1

a coupling agent with a first functional group covalently attached to the substrate of the fluidic channel and a second imide functional group covalently attached to a polymer

Methodology Applied
Scientific EffectCovalent bonding: Chemical Bonding

Implementation Method 2

covalently attached to a polymer of formula (I)... covalently attached to the substrate of the fluidic channel

Methodology Applied
Scientific EffectCovalent attachment: Chemical Bonding

Data Source

PatentEP3833696B1Flow cells with stable polymer coating and their uses for gene sequencing
Publication Date: 2022.09.14 CORNING INC
  • EP3833696B1 patent drawingFigure 1
  • EP3833696B1 patent drawingFigure 2
  • EP3833696B1 patent drawingFigure 3

AI summary

A flow cell article is provided where the flow cell article includes a substrate having one or more layers; a fluidic channel disposed in the substrate wherein the fluidic channel includes at least one reactive surface comprising: a coupling agent having a first functional group covalently attached to the substrate of the fluidic channel and a second imide functional group covalently attached to a polymer of formula (I), where R1 is a residue of an unsaturated monomer that has been copolymerized with maleic anhydride; R2 is H, an alkyl group, an oligo(ethylene glycol), and/or a dialkyl amine; m, n, and o are each from 1 to 10,000; X is a divalent NH, O, and/or S; and Z is the first functional group.