Polycation Substrate for cfDNA Extraction from Blood
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Solution Overview
Problem
Current nucleic acid extraction methods from whole blood samples often result in genomic nucleic acid contamination due to upfront lysis, which complicates the isolation of cell-free nucleic acids (cfNA) and reduces their yield.
Innovation Solution
A nucleic acid extraction material comprising a substrate with a polycation, such as a cationic copolymer of quaternized 1-vinylimidazole and N-vinylpyrrolidone, bonded to its surface, which selectively captures and elutes cell-free deoxyribonucleic acids (cfDNA) from whole blood samples with minimal genomic nucleic acid contamination.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If upfront lysis is performed to release nucleic acids from cells, then total nucleic acid yield is improved, but genomic nucleic acid contamination increases
Solution Approach 1:
The invention extracts only the desired cell-free nucleic acids from the sample while leaving cellular material intact. The polycation selectively binds cfDNA and cfRNA in the supernatant without requiring cell lysis, thereby extracting the target analyte while excluding genomic contamination.
Solution Approach 2:
The invention applies different treatment conditions to different components of the sample. The polycation is applied to the supernatant under specific conditions (pH 4.0-6.0, 0.1-1.0 mg/mL concentration) that create selective binding affinity for cell-free nucleic acids while maintaining cellular integrity and preventing genomic DNA release.
2Object-generated harmful factors
If selective capture of cell-free nucleic acids is achieved without lysis, then genomic contamination is reduced, but capture efficiency may be worsened
Solution Approach 1:
The invention optimizes multiple parameters to enhance selective capture: polycation concentration (0.1-1.0 mg/mL), pH (4.0-6.0), incubation time (5-30 minutes), and temperature (4°C to room temperature). These parameter combinations maximize cfNA binding efficiency while maintaining selectivity against cellular nucleic acids.
Solution Approach 2:
The invention uses a copolymer consisting of quaternized 1-vinylimidazole and N-vinylpyrrolidone units. This composite structure combines the cationic charge density of quaternized imidazole for strong nucleic acid binding with the hydrophilic properties of N-vinylpyrrolidone for enhanced solubility and reduced non-specific binding, achieving both efficiency and selectivity.
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 method effectively increases the yield of cfDNA from whole blood samples by reducing genomic nucleic acid contamination and allows for efficient elution and processing of cfNA, enhancing diagnostic applications such as non-invasive prenatal testing and cancer detection.
Implementation Method 1
a polycation, such as a cationic copolymer of quaternized 1-vinylimidazole and N-vinylpyrrolidone, bonded to its surface, which selectively captures and elutes cell-free deoxyribonucleic acids (cfDNA)
Implementation Method 2
which selectively captures and elutes cell-free deoxyribonucleic acids (cfDNA) from whole blood samples with minimal genomic nucleic acid contamination
Data Source
Figure 1A~1C
Figure 2~3
Figure 4
AI summary
An example of a nucleic acid extraction material includes a substrate. The nucleic acid extraction material also includes a polycation bonded to at least a portion of a surface of the substrate. The polycation consists of a polymer of a quaternized monomer selected from the group consisting of a quaternized 1-vinylimidazole monomer and a quaternized dimethylaminoethyl methacrylate monomer, or a copolymer of a neutral monomer and the quaternized monomer.