Matrices and media for storage and stabilization of biomolecules

• US 8,283,165 B2
• Filed: 09/14/2009
• Issued: 10/09/2012
• Est. Priority Date: 09/12/2008
• Status: Active Grant

• Current Assignee: GenTegra, LLC (Genisyss, LLC)
• Original Assignee: GenVault Corp. (Thermo Fisher Scientific Incorporated)
• Inventors: Hogan, Michael, Saghbini, Michael, Shi, Chunnian, Wong, David
• Primary Examiner(s): Horlick, Kenneth R.
• Assistant Examiner(s): THOMAS, DAVID C
• Application Number:
  US12/559,463
• Publication Number:
  US 20100248363A1
• Time in Patent Office: 1,121 Days
• Field of Search: None
• US Class Current: 435/374
• CPC Class Codes: C07H 21/02 with ribosyl as saccharide radical
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C09K 15/30 containing heterocyclic ring with at least one nitrogen atom as ring member
• CPC Class Codes: C12N 15/10 Processes for the isolation, preparation or purification of DNA or RNA chemical preparation of DNA or RNA C07H21/00; preparation of non-structural polynucleotides from microorganisms or with enzymes C12P19/34
• CPC Class Codes: C12N 5/0632 Cells of the oral mucosa
• CPC Class Codes: C12N 5/0634 Cells from the blood or the immune system
• CPC Class Codes: C12N 9/96 Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: Y10T 428/139 Open-ended, self-supporting conduit, cylinder, or tube-type article
• CPC Class Codes: Y10T 428/1397 Single layer [continuous layer]

Abstract

The present invention provides compositions useful for biomolecule storage comprising a water soluble inorganic compound, a stabilizer, or a combination thereof. The present invention also provides methods of using the compositions of the invention to store biomolecules in the dry state and in solution, as well as sample carriers and kits comprising compositions of the invention.

Claims

1. A dry-state matrix comprising:

boric acid or a corresponding salt of boric acid;

a plasticizer, wherein the plasticizer is a poly-alcohol;

a hydroxyl radical scavenger; and

an RNase inhibitor,

wherein the matrix is in a solid or dry form which can be converted to a liquid form by suspending, re-hydrating, or solubilizing the matrix in water.

2. The matrix of claim 1, further comprising a metal chelator.

3. The matrix of claim 1, wherein said matrix further comprises a singlet oxygen quencher selected from the group consisting of an alkyl imidazole, an indole, a sulfur-containing amino acid, a phenolic compound, an aromatic acid, azide, tocopherol, vitamin E derivatives, carotene, vitamin A derivatives, and any combination thereof.

4. The matrix of claim 1, wherein said matrix further comprises a metal chelator selected from the group consisting of EDTA, EGTA, o-phenanthroline, citrate, and any combination thereof.

5. The matrix of claim 1, wherein said hydroxyl radical scavenger is selected from the group consisting of azide, dimethyl sulfoxide, histidine, mannitol, and any combination thereof.

6. The matrix of claim 1, wherein said matrix further comprises a hydroperoxide removing agent selected from the group consisting of catalase, pyruvate, glutathione, glutathione peroxidases, and any combination thereof.

7. The matrix of claim 1, wherein said matrix is inert with respect to methods of nucleic acid amplification, nucleic acid digestion, and/or protein digestion.

8. The matrix of claim 1, further comprising a sample comprising biomolecules.

14. A sample carrier comprising:

a container, and a sample node, wherein said sample node is supported by said container and comprises a dry-state matrix of claim 1 or an aqueous medium of claim 9.

15. The sample carrier of claim 14, wherein said sample node comprises about 10 to about 1000 micrograms of said matrix, or about 10 to about 1000 microliters of said aqueous medium.

16. The sample carrier of claim 14, wherein said sample node comprises an identifying indicia.

17. The sample carrier of claim 14, further comprising a plurality of said containers, and a plurality of said sample nodes, wherein each of said plurality of sample nodes is supported by one of said plurality of said containers.

18. A kit comprising:

a container containing a dry-state matrix of claim 1, or an aqueous medium of claim 9, or a sample carrier of claim 14; and

an instruction for using said dry-state matrix, said aqueous medium, or said sample carrier for storage of biomolecules.

19. A method of storing biomolecules in a dry state, said method comprising:

mixing a sample comprising biomolecules with a dry-state matrix of claim 1 to form a mixture, and drying said mixture to store said biomolecules in a dry state.

20. The method of claim 19, wherein said sample is a liquid sample, and wherein said liquid sample is mixed with said dry-state matrix.

21. The method of claim 19, wherein said biomolecules are stored at about 25°

C. to about 72°

C.

27. The dry-state matrix of claim 1, further comprising leucine.

28. The dry-state matrix of claim 1, wherein:

the plasticizer comprises dextran;

the hydroxyl scavenger comprises mannitol; and

the RNase inhibitor comprises aluminon.

29. The dry-state matrix of claim 28, further comprising a metal chelator.

30. The dry-state matrix of claim 29, wherein the metal chelator comprises EDTA.

31. The dry-state matrix of claim 30, wherein the metal chelator further comprises citrate.

32. The dry-state matrix of claim 28, further comprising leucine.

33. The dry-state matrix of claim 30, further comprising leucine.

34. The dry-state matrix of claim 1, wherein the dry-state matrix is paper free.

35. The dry-state matrix of claim 1, wherein the dry-state matrix is in a tube.

36. The dry-state matrix of claim 1, wherein the dry-state matrix is substantially free of cellulose.

37. The method of claim 19, wherein said dry-state matrix further comprises a metal chelator selected from the group consisting of EDTA, EGTA, o-phenanthroline, citrate, and any combination thereof.

38. The method of claim 19, wherein said hydroxyl radical scavenger is selected from the group consisting of azide, dimethyl sulfoxide, histidine, mannitol, and any combination thereof.

39. The method of claim 19, wherein said dry-state matrix further comprises a hydroperoxide removing agent selected from the group consisting of catalase, pyruvate, glutathione, glutathione peroxidases, and any combination thereof.

41. The dry-state matrix of claim 1, wherein the matrix is substantially free of a biomolecule.

42. The dry-state matrix of claim 1, wherein the poly-alcohol is a polysaccharide.

9. An aqueous medium comprising:

boric acid or a corresponding salt of boric acid;

a plasticizer, wherein the plasticizer is a poly-alcohol;

a hydroxyl radical scavenger; and

an RNase inhibitor,

wherein said aqueous medium is in a liquid state.

10. The medium of claim 9, further comprising a metal chelator.

11. The medium of claim 9, wherein the RNase inhibitor is selected from the group consisting of 2′

-cytidine monophosphate free acid (2′

-CMP), aluminon, adenosine 5′

-pyrophosphate, 5′

-diphosphoadenosine 3′

-phosphate (ppA-3′

-p), 5′

-diphosphoadenosine 2′

-phosphate (ppA-2′

-p), Leucine, poly-L-aspartic acid, tyrosine-glutamic acid polymer, oligovinysulfonic acid, and 5′

-phospho-2′

-deoxyuridine 3′

-pyrophosphate P′

→

5′

-ester with adenosine 3′

-phosphate (pdUppAp).

12. The medium of claim 9, further comprising a reducing agent, a chelator, a detergent, a chaotrope, or any combination thereof.

13. The medium of claim 9, further comprising a sample comprising biomolecules.

22. A method of stabilizing biomolecules in aqueous solution, said method comprising:

mixing a sample comprising biomolecules with an aqueous medium of claim 9 to form a mixture.

23. The method of claim 22, wherein said sample is carried by a solid support.

24. The method of claim 22, wherein said sample is a solid tissue sample.

25. The method of claim 22, wherein biomolecules stabilized by said method include RNA.

26. The method of claim 22, wherein said mixture is maintained at about 4°

C. to about 37°

C.

40. The medium of claim 9, wherein said medium is inert with respect to methods of nucleic acid amplification, nucleic acid digestion, and/or protein digestion.

43. The medium of claim 9, wherein the poly-alcohol is a polysaccharide.