Charged Particle Coatings for Mild Analyte Elution
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Solution Overview
Problem
Existing methods for purifying analytes from body fluids, such as DNA extraction, often require high salt or pH conditions that are not compatible with downstream analysis techniques, necessitating additional steps like dilution or desalting, which are disadvantageous.
Innovation Solution
A method involving solid particles coated with a molecule having an opposite electric charge to the analyte, capturing the analyte through electrostatic interaction, followed by releasing the analyte by breaking down or destabilizing the coating using enzymes, reducing agents, or competitive agents, allowing elution under mild conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If high salt or pH conditions are used to elute analyte from solid particles, then the analyte can be released from the coating, but the eluted analyte becomes incompatible with downstream analysis techniques requiring additional dilution or desalting steps
Solution Approach 1:
The invention changes the elution mechanism from chemical (high salt/pH) to physical (mechanical disruption of coating). By altering how the analyte is released - through mechanical forces that disrupt the coating structure rather than chemical conditions - the analyte can be eluted in a buffer-compatible state, eliminating the need for downstream desalting or dilution steps
Solution Approach 2:
The invention extracts the charge carriers (charged groups) from the particle surface by disrupting the coating structure. This removal of the charged coating layer eliminates the electrostatic interaction holding the analyte, allowing release without introducing incompatible chemical conditions. The charge carriers are taken out physically rather than chemically neutralized
2Ease of operation
If high salt conditions are used for analyte elution, then the analyte can be released from the coating, but the salt must be removed or adjusted before the sample can be used for downstream analysis
Solution Approach 1:
The invention extracts the charge carriers from the particle surface by disrupting the coating structure. This removal of the charged coating layer eliminates the electrostatic interaction holding the analyte, allowing release without introducing incompatible chemical conditions. The charge carriers are taken out physically rather than chemically neutralized
Solution Approach 2:
The invention introduces an intermediary mechanism (coating disruption) between the analyte binding and release steps. Instead of directly using high salt to compete for binding, the coating is disrupted as an intermediary step, which indirectly causes analyte release without introducing salt or other incompatible substances into the final sample
3Measurement precision
If conventional ion exchange methods are used with silica beads, then DNA can be captured and purified, but the elution requires high salt or pH conditions that are not ideal for downstream PCR and sequencing
Solution Approach 1:
The invention changes the elution mechanism from chemical (high salt/pH) to physical (mechanical disruption of coating). By altering how the analyte is released - through mechanical forces that disrupt the coating structure rather than chemical conditions - the analyte can be eluted in a buffer-compatible state, eliminating the need for downstream desalting or dilution steps
Solution Approach 2:
The invention uses a composite structure consisting of solid particles with a coating layer. The coating material is specifically designed to provide reversible binding through electrostatic interactions while allowing mechanical disruption. This composite approach combines the benefits of strong binding during capture with gentle, compatible release conditions for downstream analysis
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 direct downstream analysis without intermediate steps, reducing the risk of analyte loss or bias, and saving time and money by eliminating the need for high salt or pH adjustments.
Implementation Method 1
contacting the sample with solid particles coated with a molecule having an electric charge opposite to the electric charge of the analyte to allow the coating to capture the analyte by electrostatic interaction
Implementation Method 2
the coating is a polyelectrolyte polymer that is broken down or destabilized by means of an enzyme
Implementation Method 3
Addition of a reducing agent results in conversion of the metal ion to an oxidation state where it is more labile and hence easily removed by chelating molecules in solution
Data Source
Figure 1A~1E
Figure 2~3
Figure 4
AI summary
The present invention relates to a method for isolating an electrically charged analyte from a body sample, comprising: a) contacting the sample with solid particles coated with a molecule having an electric charge opposite to the electric charge of the analyte to allow the coating to capture the analyte by electrostatic interaction; b) separating the solid particles and the sample; and c) releasing the analyte from the solid particles by breaking down or destabilizing the coating. When the coating is a polymer it is broken down or destabilized by means of an enzyme. When the coating comprises a metal ion that is chelated to the surface of the solid particles, the coating is destabilized or broken down by means of a reducing agent. The coating can also be hydrophobic comprising an amphiphilic molecule embedded therein and then the breaking down or destabilizing the coating is achieved by means of a detergent or a competitive agent.