Methods for electroelution of biomolecules
View Patent ↗A method of eluting biomolecules, such as nucleic acids from a biological sample by electroelution is provided. An example of a method includes various steps, such as loading the biological sample to a device comprising a housing, at least two conductive redox polymer electrodes operationally coupled to the housing and a biomolecule impermeable layer disposed on at least one of the electrodes. The loading of sample is followed by initiating an electrical connection to generate an electric field strength sufficient to elute biomolecules from the biological sample; and eluting the biomolecules from the biological sample.
1. A method of eluting biomolecules from a biological sample, comprising:
loading the biological sample to a device comprising a housing, at least two conductive redox polymer electrodes operationally coupled to the housing and a biomolecule impermeable layer disposed on at least one of the electrodes;
initiating an electrical connection to generate an electric field strength sufficient to elute biomolecules from the biological sample, wherein the electric field strength is at least about 400 V/cm; and
eluting the biomolecules from the biological sample, wherein eluting the biomolecules comprises electrophoretically eluting the biomolecules from the biological sample, wherein the at least two conductive redox polymer electrodes comprise conductive redox polymers of 0.05 μg-0.5 mg per mm 2 of electrode surface and the conductive redox polymer is cross-linked.
2. The method of claim 1 , wherein the conductive redox polymer is selected from polyacetylenes, polyphenylene vinylenes, polypyrroles, polyThiophenes, polyanilines, polyphenylene sulfide, or polyfluorenes.
3. The method of claim 1 , wherein the conductive redox polymer comprises cross-linked Pedot-PSS and the biomolecule impermeable layer comprises a sulfonated tetrafluoroethylene based fluoropolymer-copolymer.
4. The method of claim 1 , wherein the loading of the biological sample is effected by direct loading of the biological sample to the housing, loading of a biological sample laden substrate into the housing, loading of the biological sample in a package configured to release the sample during operation, or combinations thereof.
5. The method of claim 1 , further comprising wetting the electrodes and the biomolecule impermeable layer.
6. The method of claim 1 , further comprising electrophoretically driving the eluted biomolecules to a surface of the biomolecule impermeable layer.
7. The method of claim 1 , further comprising releasing the biomolecules from the surface of the biomolecule impermeable layer by reversing an ion-conduction between the electrodes.
8. The method of claim 7 , further comprising eluting impurities from the housing.
9. The method of claim 1 , wherein the conductive redox polymer comprises Pedot-PSS.
10. The method of claim 1 , wherein the housing comprises a cylindrical cartridge, a microfluidic channel or combinations thereof.
11. The method of claim 1 , wherein the biomolecule impermeable layer comprises cellulose, sulfonated tetrafluoroethylene based fluoropolymer-copolymer or combinations thereof.
12. The method of claim 1 , wherein the biomolecules are nucleic acids.
13. The method of claim 1 , wherein the biological sample comprises a blood, a buccal swab, a body fluid, a serum, a sputum, an excretory product, a cell-extract, a cell, a tissue, a liquid comprising nucleic acids or combinations thereof.
14. The method of claim 1 , further comprising wetting the electrodes by adding an electrolyte to the housing.
15. A method of eluting nucleic acids from a biological sample, comprising:
loading the biological sample to a device comprising a housing, at least two conductive redox polymer electrodes operationally coupled to the housing and a biomolecule impermeable layer disposed on at least one of the electrodes;
initiating an electrical connection to generate an electric field strength sufficient to elute nucleic acids from the biological sample, wherein the electric field strength is at least about 25 V/cm; and
eluting the nucleic acids from the biological sample, wherein eluting the nucleic acids comprises electrophoretically eluting the nucleic acids from the biological sample, wherein the at least two conductive redox polymer electrodes comprise conductive redox polymers of 0.05μg-0.5 mg per mm 2 of electrode surface and the conductive redox polymer is cross-linked.
16. The method of claim 15 , wherein the conductive redox polymer comprises cross-linked Pedot-PSS and the biomolecule impermeable layer comprises a sulfonated tetrafluoroethylene based fluoropolymer-copolymer.
17. The method of claim 15 . further comprising wetting the electrodes and the biomolecule impermeable layer.
18. The method of claim 15 , further comprising electrophoretically driving the eluted biomolecules to a surface of the biomolecule impermeable layer.
19. The method of claim 15 , further comprising releasing the biomolecules from the surface of the biomolecule impermeable layer by reversing an ion-conduction between the electrodes.
20. The method of claim 19 , further comprising eluting impurities from the housing.