Reversible Silane Surface Functionalization for Flow Cell Renewal
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Solution Overview
Problem
Current sequencing technologies, such as sequencing-by-synthesis (SBS), require a new flow cell after each use due to the irreversible attachment of silane layers, leading to high costs and waste, as there is no efficient method to renew or recycle the flow cell surfaces.
Innovation Solution
The use of reversible interactions, including host-guest complexes and copper-imine bonds, allows for the reversible immobilization of biological molecules like DNA on silane-functionalized surfaces, enabling the reuse of flow cells by dissociating the molecules under specific conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of repair
If a strong base is used to remove the silane layer, then the flow cell can be renewed, but a CVD oven is required which increases device complexity and cost
Solution Approach 1:
The patent extracts the silane layer removal process from the complex CVD oven system by using a simple base treatment that can be performed without specialized equipment. The base selectively removes the silane layer while leaving the flow cell substrate intact, enabling renewal through a straightforward chemical treatment rather than requiring removal and re-deposition equipment.
Solution Approach 2:
The patent changes the chemical parameters of the silane layer by treating it with a base solution, which alters the silane's chemical state from a stable covalently-bonded layer to a removable form. This parameter change enables the silane to be detached under mild conditions without requiring high-temperature CVD equipment, thus simplifying the renewal process.
2Reliability
If the flow cell is disposed after each use, then sequencing quality is maintained, but cost and waste increase
Solution Approach 1:
The patent implements a recovery process where the valuable flow cell substrate is preserved and the only discarded material is the spent silane layer. By selectively removing and discarding only the consumable silane component while recovering and reusing the expensive flow cell substrate, the system maintains sequencing quality through proper surface preparation while dramatically reducing waste and cost.
3Strength
If covalent attachment is used to attach DNA to the surface, then immobilization strength is improved, but reversibility is lost
Solution Approach 1:
The patent creates a dynamic system where the attachment strength can be modulated. The silane layer provides strong initial immobilization of DNA through covalent bonds, but the entire silane layer can be reversibly removed by base treatment. This dynamic approach allows the system to switch between strong attachment (during sequencing) and complete release (for renewal), providing both strength and reversibility at different operational stages.
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 enables the multiple use of flow cells, reducing waste and costs by allowing the regeneration of the flow cell surface while maintaining the integrity of the functionalized silane layer, thus facilitating cost-effective and sustainable DNA sequencing processes.
Implementation Method 1
it is known that a strong base can be used to efficiently remove the silane layer
Data Source
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Figure 3A~3B
AI summary
Some embodiments described herein relate to a substrate comprising a silane functionalized surface for reversibly immobilizing a biological molecule of interest, such as oligonucleotides, polynucleotides, or protein. Methods for immobilizing the biological molecule and the use in DNA sequencing and other diagnostic applications are also disclosed.