Dry-State Nucleic Acid Substrate with E-Beam Modified Surface
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Solution Overview
Problem
Existing methods for extracting biomolecules from solid substrates often result in low recovery and degradation of nucleic acids due to the use of extreme environmental factors like pH, temperature, and salt concentrations, which can cause denaturation and degradation of RNA and DNA.
Innovation Solution
A solid substrate modified with negatively charged groups via e-beam modification, impregnated with protein denaturing agents and reducing agents, allows for the stabilization and effective elution of nucleic acids in a dry state, preserving their integrity and enhancing recovery through a method that involves binding, washing, and elution at physiological pH.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If extreme environmental factors (pH, temperature, salt concentrations) are used to elute nucleic acids from substrates, then biomolecules can be released from the substrate, but nucleic acid stability deteriorates and degradation occurs
Solution Approach 1:
The invention changes the chemical parameters of the substrate surface by introducing negatively charged groups (carboxyl, sulfonate, phosphate) that enable nucleic acid binding at physiological pH. This eliminates the need for extreme pH changes, high salt concentrations, or high temperature treatments that would degrade the nucleic acids, thereby maintaining both high recovery and stability
Solution Approach 2:
The negatively charged substrate surface acts as an intermediary that facilitates nucleic acid binding and release through electrostatic interactions. This intermediary mechanism allows for gentle elution conditions using mild buffers at physiological pH, avoiding the need for harsh chemicals or extreme conditions that would compromise nucleic acid integrity
2Quantity of substance
If conventional extraction solvents are used to extract biomolecules from substrates, then biomolecules can be recovered, but biomolecule stability and downstream usability are adversely affected
Solution Approach 1:
The invention extracts the harmful extraction solvents from the process by using a substrate-based binding approach. Nucleic acids are bound directly to the negatively charged substrate surface and then eluted using mild buffers, eliminating the need for organic solvents or harsh chemicals that would compromise biomolecule usability in downstream applications
3Ease of operation
If high temperature or alkaline pH is used to release bound biomolecules from substrate, then biomolecules can be eluted through denaturation, but nucleic acid structural integrity is compromised
Solution Approach 1:
Instead of using high temperature or alkaline pH to denature and release biomolecules, the invention inverts the approach by using negatively charged substrate surfaces that bind nucleic acids through electrostatic attraction at physiological pH. Elution is achieved by reversing the charge interaction through simple buffer exchange, maintaining nucleic acid structural integrity while achieving efficient release
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
The described method achieves higher DNA recovery and maintains the structural and functional integrity of nucleic acids, reducing the need for high ionic strength or alkaline pH buffers, thus minimizing degradation and improving downstream applications.
Implementation Method 1
a surface modified with a plurality of negatively charged groups at physiological pH grafted by e-beam modification
Implementation Method 2
binding the biomolecule to the surface
Implementation Method 3
one or more protein denaturing agents impregnated therein the substrate in a substantially dry state
Data Source
AI summary
A solid substrate for biological sample storage under dry-state and elution of biomolecules is provided. The dry, solid substrate comprises a surface modified with a plurality of hydrophilic groups; and the substrate is comprised of one or more protein denaturing agents impregnated therein under a substantially dry state. A method for elution of biomolecules from biological samples is also provided. The compositions disclosed herein provide for enhanced elution and recovery of biomolecules, such as nucleic acids, from the sample. The sample is disposed on a substrate, dried to a substantially dry state; eluted from the biological sample dried on the substrate by rehydrating the substrate in an elution buffer.