Nucleic Acid Binding Support for High-Yield miRNA Purification
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Solution Overview
Problem
Current methods for isolating and purifying extracellular nucleic acids, such as miRNAs, from body fluids like blood, plasma, and serum are inefficient, resulting in low yields and difficulties in processing large sample volumes due to challenges in binding and blocking issues with nucleic acid-binding supports at high alcohol concentrations.
Innovation Solution
A method involving the use of a nucleic acid-binding support material with chaotropic compounds, at least two different detergents, and a branched or unbranched alcohol at concentrations ≥40% (v/v) for efficient binding of nucleic acids, allowing for high-yield purification of short-chain nucleic acids like miRNAs from large sample volumes without blocking small-area membranes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If high alcohol concentration (≥40% v/v) is used during binding, then the yield of short-chain nucleic acids is markedly increased, but the sample fluid blocks small-area nucleic acid binding supports such as membranes
Solution Approach 1:
The patent introduces an intermediary substance (chaotropic compound) that mediates between the high alcohol concentration and the membrane surface. This chaotropic compound prevents direct interaction between alcohol and membrane components that would cause blockage, while still allowing the high alcohol concentration to maintain nucleic acid binding efficiency. The intermediary substance acts as a protective layer that enables both high yield and prevents harmful blockage effects.
2Productivity
If standard purification methods are used, then the process is simple, but the yield of extracellular nucleic acids is comparatively low
Solution Approach 1:
The patent fundamentally changes the chemical parameters of the binding buffer by incorporating chaotropic compounds at specific concentrations. This parameter change transforms the binding mechanism to specifically enhance short-chain nucleic acid binding while maintaining compatibility with membrane surfaces. The modified chemical environment enables high yield recovery without requiring complex additional purification steps, thus improving productivity without significantly increasing operational complexity.
3Measurement precision
If large sample volumes are processed, then the detection sensitivity for low concentrations of nucleic acids is improved, but the processing time and complexity increase
Solution Approach 1:
The patent modifies the binding buffer parameters with chaotropic compounds that enhance nucleic acid binding efficiency. This parameter change allows large sample volumes to be processed more rapidly and efficiently, as the improved binding characteristics reduce the need for extended processing steps. The enhanced binding affinity enables faster separation and concentration of nucleic acids from large volumes, thereby improving detection sensitivity while minimizing time loss.
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 method significantly increases the yield of short-chain nucleic acids, particularly miRNAs, by preventing membrane blockage and enabling effective processing of large sample volumes, resulting in a high concentration effect and improved sensitivity for detecting low concentrations of nucleic acids.
Implementation Method 1
contacting the starting material with said nucleic acid-binding support material in the presence of at least one chaotropic compound
Implementation Method 2
at least two different detergents
Implementation Method 3
binding the nucleic acids to a nucleic acid-binding support material by contacting the starting material with said nucleic acid-binding support material
Implementation Method 4
with the concentration of said alcohol being ≧40% (v/v)
Data Source
AI summary
The invention describes a method of and kits for isolating and/or purifying nucleic acids, more specifically short-chain nucleic acids such as miRNA, from a nucleic acid-containing starting material, characterized by the following method steps of:(a) binding the nucleic acids to a nucleic acid-binding support material by contacting the starting material with said nucleic acid-binding support material in the presence of at least one chaotropic compound, at least two different detergents and at least one branched and/or unbranched alcohol, preferably isopropanol, with the concentration of said alcohol being 40% (v/v);(b) optionally eluting the bound nucleic acids from the nucleic acid-binding support material.The method of the invention is particularly suitable for purifying circulating, extracellular miRNA from blood.