Fabrication of hierarchical silica nanomembranes and uses thereof for solid phase extraction of nucleic acids

Patent metadata

Field	Value
Publication number	US-9765325-B2
Application number	US-201414341957-A
Country	US
Kind code	B2
Filing date	Jul 28, 2014
Priority date	Aug 5, 2013
Publication date	Sep 19, 2017
Grant date	Sep 19, 2017

Abstract

The present invention provides a novel method to fabricate silica nanostructures on thin polymer films based on silica deposition and self-wrinkling induced by thermal shrinkage. These micro- and nano-scale structures have vastly enlarged the specific area of silica, thus the silica nanomembranes can be used for solid phase extraction of nucleic acids. The inventive silica nanomembranes are suitable for nucleic acid purification and isolation and demonstrated better performance than commercial particles in terms of DNA recovery yield and integrity. In addition, the silica nanomembranes have extremely high nucleic acid capacity due to its significantly enlarged specific surface area of silica. Methods of use and devices comprising the silica nanomembranes are also provided.

First claim

The invention claimed is:

1. A silica nanomembrane comprising a heat shrunken polymer core coated with a silicon dioxide layer, wherein a surface structure comprises microscale wrinkles structures overlaid with discreet, substantially-planar nanoscale flakes having a length from about 10 nm to about 10 μm.

2. The silica nanomembrane of claim 1 , wherein the silicon dioxide layer has a thickness of between about 2 nm to about 500 nm.

3. The silica nanomembrane of claim 1 , wherein the polymer core has a shrunken thickness of between about 5 μm and 5 mm.

4. The silica nanomembrane of claim 1 , wherein the polymer core has a pre-shrunken thickness of between about 5 μm and 500 μm.

5. The silica nanomembrane of claim 1 , wherein the polymer core is planar.

6. The silica nanomembrane of claim 1 , wherein the polymer core is a thermoplastic material selected from the group consisting of polyolefins, polystyrenes, polycarbonates, polymethyl methacrylate, polyvinyl chloride, polyethylene, fluorinated ethylene propylene, polytetrafluoroethylene, and polyvinylidene fluoride.

7. The silica nanomembrane of claim 1 , wherein the silica nanomembrane has a square or circular shape.

8. The silica nanomembrane of claim 1 , wherein the silicon dioxide layer is derivatized.

9. The silica nanomembrane of claim 8 , wherein the silicon dioxide layer is derivatized with aminopropyl groups, chloropropyl groups, octadecyl groups, octyl groups, quaternary ammonium groups, diethlylaminoethyl group, sulfonic acid groups, phenyl groups, biotin, streptavidin, antibodies, or enzymes.

10. A method for making a silica nanomembrane comprising:
a) depositing onto a polymer film or core having an original size, a layer of silicon dioxide; and
b) heating the composition of a) at a sufficient temperature and time to allow the polymer film or core to shrink, and
wherein the shrinking of the polymer film or core creates silica microstructures and/or nanostructures on the surface of the layer of silica on the silica nanomembrane wherein the silica nanomembrane comprise a surface structure of microscale wrinkle structures overlaid with discreet, substantially planar nanoscale flakes having a length from about 10 nm to about 10 um.

11. The method of claim 10 , wherein the polymer film or core shrinks to between about 0.1% to about 75% of its original size.

12. The method of claim 10 , wherein the polymer film or core is planar.

13. The method of claim 10 , wherein the polymer core has a pre-shrunken thickness of between about 5 μm and 500 μm in thickness.

14. The method of claim 10 , wherein the polymer core is a thermoplastic which is heat-shrinkable and selected from the group consisting of polyolefins, polystyrenes, polycarbonates, polymethyl methacrylate, polyvinyl chloride, polypropylene, polyethylene, fluorinated ethylene propylene, polytetrafluoroethylene, and polyvinylidene fluoride.

15. The method of claim 10 , wherein the silicon dioxide layer has a thickness of between about 2 nm to about 500 nm.

16. The method of claim 10 , wherein the silicon dioxide layer is deposited on the thermoplastic core by chemical vapor deposition, electrophoretic deposition, dip-coating, physical vapor deposition, electron beam vapor deposition, sputtering, spin-coating, or liquid phase deposition.

17. The method of claim 10 , at step b) wherein the composition is heated at a temperature between 100° F. and 500° F.

18. The method of claim 17 , wherein the temperature is 250° F.

19. The method of claim 10 , at step b) wherein the composition is heated for between 10 sec to 10 minutes.

20. The method of claim 19 , wherein the composition is heated for 3 minutes.

21. The method of claim 16 , wherein silica structures comprise microscale wrinkles and nanoscale flakes.

22. A method for extracting nucleic acids from a sample comprising:
a) contacting a sample comprising nucleic acids with a sufficient amount of silica nanomembranes of claim 1 ;
b) allowing the nucleic acids in the sample to adsorb onto the silica nanomembranes;
c) washing the silica nanomembranes to remove any non-nucleic acid components; and
d) desorbing the nucleic acids from the silica nanomembranes to obtain the isolated and purified nucleic acids from the sample.

23. The method of claim 22 , wherein the sample is from a cell or a tissue.

24. The method of claim 23 , further comprising contacting the sample with a lysis and/or digestion solution prior to step a), followed by one or more washing steps to remove cellular debris and lysis and/or digestion components.

25. The method of claim 22 , wherein at step a) the nucleic acids are contacted with a chaotropic agent.

26. The method of claim 22 , further comprising at step b) contacting the sample with a sufficient amount of silica nanomembranes in the presence of an aqueous alcoholic solution.

27. The method of claim 22 , wherein step d) comprises two or more washes.

28. The method of claim 22 , wherein step d) comprises subsequently adding to the sample a DNAse or an RNAse to further enrich the sample with RNA or DNA.

29. The method of claim 22 , further comprising a drying step after step d).

30. The method of claim 22 , wherein at step e) the desorption of the nucleic acids from the silica nanomembranes is by application of an elution solution.

31. The method of claim 22 , wherein the nucleic acids are DNA.

32. The method of claim 22 , wherein the nucleic acids are RNA.

33. The method of claim 22 , wherein the nucleic acids are from plasmid, genomic, mitochondrial, vesicles, or cell free sources.

34. A method for extracting nucleic acids from formalin fixed paraffin embedded (FFPE) samples comprising:
a) deparaffinizing a FFPE sample comprising nucleic acids;
b) contacting the sample with a sufficient amount of silica nanomembranes of claim 1 ;
c) allowing the nucleic acids in the sample to adsorb onto the silica nanomembranes;
d) washing the silica nanomembranes to remove any non-nucleic acid components; and
e) desorbing the nucleic acids from the silica nanomembranes to obtain the isolated and purified nucleic acids from the sample.

35. The method of claim 34 , wherein the sample is from a cell or a tissue.

36. The method of claim 34 , wherein the deparaffinization step b) comprises contacting the sample with an organic solvent.

37. The method of claim 36 , wherein the organic solvent is xylene, mineral oil, hexadecane, toluene, 5-chloro-2-methyl-4-isothiazolin-3-one, 5-chloro-2-methyl-4-isothiazolin-3-one; a terpene or isoparaffinic hydrocarbon, and 2-butoxyethanol.

38. The method of claim 36 , wherein step b) further comprises removing the organic solvent, and washing the sample.

39. The method of claim 34 , further comprising contacting the sample with a lysis and/or digestion solution prior to step c), followed by one or more washing steps to remove cellular debris and lysis and/or digestion components.

40. The method of claim 34 , wherein at step c) the nucleic acids are contacted with a chaotropic agent.

41. The method of claim 34 , further comprising at step c) contacting the sample with a sufficient amount of s

Assignees

• Univ Johns Hopkins

Inventors

• Wang Tza-Huei
• Zhang Yi

Classifications

• B01D63/087

  Single membrane modules · CPC title

• B01D69/02

  characterised by their properties · CPC title

• B01D2325/06

  Surface irregularities · CPC title

• B01J20/261

  obtained by reactions only involving carbon to carbon unsaturated bonds (macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds per se C08F) · CPC title

• B01D67/0083

  Thermal after-treatment · CPC title