MATRICES AND MEDIA FOR STORAGE AND STABILIZATION OF BIOMOLECULES

• US 20100248363A1
• Filed: 09/14/2009
• Published: 09/30/2010
• Est. Priority Date: 09/12/2008
• Status: Active Grant

• Current Assignee: GenTegra, LLC (Genisyss, LLC)
• Original Assignee: GenVault Corp. (Thermo Fisher Scientific Incorporated)
• Inventors: Wong, David, Shi, Chunnian, Hogan, Michael, Saghbini, Michael
• Granted Patent: US 8,283,165 B2
• Time in Patent Office: Days
• 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:

an inorganic compound; and

a stabilizer, a plasticizer, or a combination thereof,

wherein the stabilizer is selected from the group consisting of a singlet oxygen quencher, a hydroxyl radical scavenger, a hydroperoxide removing agent, a reducing agent, a metal chelator, a detergent, a chaotrope, or any combination thereof, and

wherein said inorganic compound is water soluble and wherein the matrix is in solid state.

2. The matrix of claim 1, wherein said inorganic compound is a metal chelator or a microcidal agent, or is inert with respect to biomolecules.

3. The matrix of claim 1, wherein said inorganic compound comprises an element from Group IIIA, Group VA, or Group VB of the Periodic Table of Elements.

4. The matrix of claim 1, wherein said inorganic compound comprises an element selected from the group consisting of boron, phosphorus, vanadium, and aluminum.

5. The matrix of claim 1, wherein said inorganic compound comprises boric acid, a corresponding salt of boric acid, phosphoric acid, a corresponding salt of phosphoric acid, a salt comprising vanadate, potassium alum, soda alum, ammonium alum, or any combination thereof.

6. The matrix of claim 1, wherein said matrix comprises a stabilizer, and wherein said stabilizer 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.

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

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

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

10. The matrix of claim 1, wherein said matrix comprises a plasticizer selected from the group consisting of monosaccharides, disaccharides, complex sugars, and linear or branched short-chain poly-alcohols.

11. 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.

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

31. 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, 13, 15, or 19, or an aqueous medium of claim 23 or 28.

32. The sample carrier of claim 31, 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.

33. The sample carrier of claim 31, further comprising an identifying indicia.

34. The sample carrier of claim 31, wherein said sample node comprises an identifying indicia.

35. The sample carrier of claim 31, 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.

36. A kit comprising:

a container containing a dry-state matrix of claim 1, 13, 15, or 19, or an aqueous medium of claim 23 or 28, or a sample carrier of claim 31; and

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

37. 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, 13, 15, or 19, or an aqueous medium of claim 23 or 28, to form a mixture, and

drying said mixture to store said biomolecules in a dry state.

38. The method of claim 37, wherein said sample is a liquid sample, and wherein said liquid sample is mixed with said dry-state matrix.

39. The method of claim 37, wherein said sample is carried by a solid support, and wherein said mixing comprises rinsing said solid support with said aqueous medium.

40. The method of claim 37, wherein said biomolecules are stored at about 25°

C. to about 72°

C.

13. A dry-state matrix comprising:

an inorganic compound; and

a sample comprising biomolecules,

wherein said inorganic compound is water soluble and wherein said matrix is in solid state.

14. The matrix of claim 13, wherein said inorganic compound comprises boric acid, a corresponding salt of boric acid, phosphoric acid, a corresponding salt of phosphoric acid, a salt comprising vanadate, potassium alum, soda alum, ammonium alum, or combinations thereof.

15. A dry-state matrix comprising:

a stabilizer comprising a singlet oxygen quencher, a hydroxyl radical scavenger, a hydroperoxide removing agent, a reducing agent, a chelator, a detergent, a chaotrope, or any combination thereof, and

an inorganic salt, a plasticizer, or any combination thereof,

wherein said matrix is in solid state.

16. The matrix of claim 15, wherein said stabilizer comprises a singlet oxygen quencher, and wherein said singlet oxygen quencher is selected from the group consisting of an alkyl imidazole, an indole, a sulfur-containing amino acid, a phenolic compound, an aromatic acid, an azide salt, tocopherol, vitamin E derivatives, carotene, vitamin A derivatives, and combinations thereof.

17. The matrix of claim 15, wherein said matrix comprises a plasticizer selected from the group consisting of monosaccharides, disaccharides, complex sugars, and linear or branched short-chain poly-alcohols.

18. The matrix of claim 15, further comprising a sample comprising biomolecules.

19. A dry-state matrix comprising:

at least three components selected from the list consisting of an inorganic compound, a singlet oxygen quencher, a hydroxyl radical scavenger, a hydroperoxide removing agent, a reducing agent, a metal chelator, a detergent, and a plasticizer,

wherein said inorganic compound is water soluble, and wherein said matrix is in solid state.

20. The matrix of claim 19, wherein said matrix comprises an inorganic compound, a hydroxyl radical scavenger, and a hydroperoxide removing agent.

21. The matrix of claim 20, further comprising a plasticizer, a metal chelator, an RNase inhibitor, or any combination thereof.

22. The matrix of claim 19, further comprising a sample comprising biomolecules.

23. An aqueous medium comprising:

an inorganic compound;

a stabilizer, a plasticizer, or a combination thereof, and

optionally, an RNase inhibitor,

wherein said stabilizer is selected from the group consisting of a singlet oxygen quencher, a hydroxyl radical scavenger, a hydroperoxide removing agent, and any combination thereof, and

wherein said inorganic compound is water soluble.

24. The medium of claim 23, wherein said inorganic compound comprises boric acid, a corresponding salt of boric acid, phosphoric acid, a corresponding salt of phosphoric acid, a salt comprising vanadate, potassium alum, soda alum, ammonium alum, or combinations thereof.

25. The medium of claim 23, comprising an RNase inhibitor 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, 5′

-phospho-2′

-deoxyuridine 3′

-pyrophosphate P′

→

5′

-ester with adenosine 3′

-phosphate (pdUppAp).

26. The medium of claim 23, further comprising a reducing agent, a chelator, a detergent, a chaotrope, or any combination thereof.

27. The medium of claim 23, further comprising a sample comprising biomolecules.

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

mixing a sample comprising biomolecules with an aqueous medium of claim 23 or 28, to form a mixture.

42. The method of claim 41, wherein said sample is carried by a solid support.

43. The method of claim 41, wherein said sample is a solid tissue sample.

44. The method of claim 41, wherein biomolecules stabilized by said method include RNA.

45. The method of claim 41, wherein said mixture is maintained at about 4°

C. to about 37°

C.

28. An aqueous medium comprising:

one or more hydroxyl radical scavengers;

an inorganic salt, a plasticizer, or any combination thereof, and

optionally, an RNase inhibitor.

29. The medium of claim 28, further comprising a singlet oxygen quencher, a reducing agent, a chelator, a detergent, a chaotrope, or any combination thereof.

30. The medium of claim 28, further comprising a sample comprising biomolecules.