Methods for sterilizing biological materials by irradiation over a temperature gradient

• US 6,908,591 B2
• Filed: 07/18/2002
• Issued: 06/21/2005
• Est. Priority Date: 07/18/2002
• Status: Expired due to Fees

• Current Assignee: Clearant, Inc.
• Original Assignee: Clearant, Inc.
• Inventors: Lynch, Tom, Calvert, Glenn, Burgess, Wilson, Drohan, William N., MacPhee, Martin, Miekka, Shirley, Mann, David, Kent, Randall S.
• Primary Examiner(s): MCKANE, ELIZABETH L
• Application Number:
  US10/197,249
• Publication Number:
  US 20040033160A1
• Time in Patent Office: 1,069 Days
• Field of Search: 422/21, 422/22, 422/24, 422/23, 435/1.1, 435/2
• US Class Current: 422/22
• CPC Class Codes: A61L 2/0029 Radiation
• CPC Class Codes: A61L 2/0035 Gamma radiation
• CPC Class Codes: A61L 2/0041 X-rays
• CPC Class Codes: A61L 2/0047 Ultraviolet radiation
• CPC Class Codes: A61L 2/0052 Visible light
• CPC Class Codes: A61L 2/0064 Microwaves
• CPC Class Codes: A61L 2/007 Particle radiation, e.g. electron-beam, alpha or beta radiation
• CPC Class Codes: A61L 2/10 Ultraviolet radiation
• CPC Class Codes: A61L 2202/22 Blood or products thereof

Abstract

Methods are disclosed for sterilizing tissue to reduce the level of one or more active biological contaminants or pathogens therein, such as viruses, bacteria, (including inter- and intracellular bacteria, such as mycoplasmas, ureaplasmas, nanobacteria, chlamydia, rickettsias), yeasts, molds, fungi, spores, prions or similar agents responsible, alone or in combination, for TSEs and/or single or multicellular parasites. The methods involve sterilizing one or more tissues with irradiation.

Claims

1. A method for sterilizing a biological material that is sensitive to radiation, said method comprising:

(a) determining the maximum acceptable temperature (T_max) for said biological material during irradiation; and

(b) irradiating said biological material with radiation for a time effective to sterilize said biological material at a rate effective to sterilize said biological material, wherein said irradiating is performed under conditions whereby the temperature of said biological material increases during said irradiating from an initial temperature (T_i) to a final temperature (T_f), wherein said initial temperature (T_i) is no greater than about T_max−

T, wherein T is about equal to the total dose of said radiation (D) divided by the specific heat capacity (c) of said biological material.

3. The method according to claim 1 or 2, wherein said final temperature (T_f) is at or below a temperature effective to protect said biological material from said radiation.

4. The method according to claim 1 or 2, wherein at least one stabilizing process is applied to said biological material prior to said irradiating, said stabilizing process being selected from the group consisting of:

(a) adding to said biological material at least one stabilizer;

(b) reducing the residual solvent content of said biological material;

(c) reducing the oxygen content of said biological material;

(d) adjusting the pH of said biological material; and

(e) adding to said biological material at least one non-aqueous solvent, wherein said stabilizing process is effective to protect said biological material from said radiation.

5. The method according to claim 1 or 2, wherein said increase in the temperature of the biological material is about 0.25°

C./kGy.

6. The method according to claim 1 or 2, wherein said increase in the temperature of the biological material is about 1.2°

C./kGy.

7. The method according to claim 1 or 2, wherein said rate is effective to protect said biological material from said radiation.

8. The method according to claim 4, wherein at least two stabilizing processes are applied to said biological material prior to said irradiating.

9. The method according to claim 1 or 2, wherein said rate is at least about 3.0 kGy/hr.

10. The method according to claim 1 or 2, wherein said rate is at least about 30.0 kGy/hr.

11. The method according to claim 1 or 2, wherein said rate is at least about 45.0 kGy/hr.

12. The method according to claim 1 or 2, wherein said biological material is maintained in a low oxygen atmosphere.

13. The method according to claim 1 or 2, wherein said biological material is maintained in an atmosphere comprising at least one noble gas or nitrogen.

14. The method according to claim 13, wherein said noble gas is argon.

15. The method according to claim 1 or 2, wherein said biological material is maintained in a vacuum.

16. The method according to claim 1 or 2, wherein at least one sensitizer is added to said biological material prior to said step of irradiating said biological material.

17. The method according to claim 1 or 2, wherein said radiation is selected from the group consisting of gamma, corpuscular, electromagnetic and combinations thereof.

18. The method according to claim 17, wherein said electromagnetic radiation is selected from the group consisting of radio waves, microwaves, visible and invisible light, ultraviolet light, x-ray radiation, gamma radiation and combinations thereof.

19. The method according to claim 1 or 2, wherein said irradiation is conducted at a temperature below ambient temperature.

20. The method according to claim 1 or 2, wherein said radiation is conducted at a temperature below the freezing point of said biological material.

21. The method according to claim 1 or 2, wherein said biological material is selected from the group consisting of dextrose, urokinase, thrombin, purified protein fraction, blood, blood cells, alpha-1 proteinase inhibitor, digestive enzymes, blood proteins and tissue.

22. The method according to claim 21, wherein said tissue is selected from the group consisting of tendons, nerves, bone, teeth, bone marrow, skin grafts, cartilage, corneas, arteries, veins, heart valves, ligaments and demineralized bone matrix.

23. The method according to claim 21, wherein said digestive enzymes are selected from the group consisting of galactosidases and sulfatases.

24. The method according to claim 21, wherein said blood proteins are selected from the group consisting of albumin, Factor VIII, Factor VII, Factor IV, fibrinogen, monoclonal immunoglobulins and polyclonal immunoglobulins.

25. The method according to claim 1 or 2, wherein said biological material is milk.

26. The method according to claim 1 or 2, wherein said biological material is serum or plasma.

27. The method according to claim 4, wherein said residual solvent content is reduced by a method selected from the group consisting of lyophilization, drying, concentration, addition of solute, evaporation, chemical extraction, spray-drying and vitrification.

28. The method according to claim 4, wherein said residual solvent content is reduced to a level less than about 33%.

29. The method according to claim 4, wherein said residual solvent content is reduced to a level less than about 15%.

30. The method according to claim 4, wherein said residual solvent content is reduced to a level less than about 8%.

31. The method according to claim 4, wherein said residual solvent is an organic solvent.

32. The method according to claim 31, wherein said organic solvent is selected from the group consisting of ethanol, isopropanol, polyethylene glycol and mixtures thereof.

33. The method according to claim 4, wherein said residual solvent is an aqueous solvent.

34. The method according to claim 4, wherein said at least one stabilizer is an antioxidant.

35. The method according to claim 4, wherein said at least one stabilizer is selected from the group consisting of:

ascorbic acid or a salt or ester thereof;

glutathione;

vitamin E or a derivative thereof;

albumin;

sucrose;

glycyiglycine;

L-carnosine;

cysteine;

silymarin;

diosmin;

hydroquinonesulfonic acid;

6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid;

uric acid or a salt or ester thereof;

methionine;

histidine;

N-acetyl cysteine;

lipoic acid;

sodium formaldehyde sulfoxylate;

gallic acid or a derivative thereof;

propyl gallate;

ethanol;

acetone;

rutin;

epicatechin;

biacalein;

purpurogallin;

mannitol;

trehalose;

DMSO and mixtures of two or more thereof.

36. The method according to claim 35, wherein said at least one stabilizer is selected from the group consisting of mannitol, propylene glycol, trehalose, DMSO and mixtures thereof.

37. The method according to claim 35, wherein said mixtures of two or more stabilizers comprise ascorbic acid or a salt or ester thereof and at least one stabilizer selected from the group consisting of:

ethanol;

acetone;

6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid;

6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid;

albumin;

sucrose;

glycylglycine;

L-camosine;

N-acetyl cysteine;

silymarin;

diosmin;

lipoic acid;

hydroquinonesulfonic acid;

sodium formaldehyde sulfoxylate; and

gallic acid or a derivative thereof.

38. The method according to claim 35, wherein said mixtures of two or more stabilizers comprise uric acid or a salt or ester thereof and at least one stabilizer selected from the group consisting of:

ascorbic acid or a salt or ester thereof;

ethanol;

acetone;

6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid;

6-hydroxy-2,5,7,8-tetraniethylchroman-2-carboxylic acid;

albumin;

sucrose;

glycylglycine;

L-camosine;

N-acetyl cysteine;

silymarin;

diosmin;

lipoic acid;

hydroquinonesulfonic acid;

sodium formaldehyde sulfoxylate; and

gallic acid or a derivative thereof.

39. The method according to claim 4, wherein said at least one stabilizer is a dipeptide stabilizer.

40. The method according to claim 4, wherein said non-aqueous solvent is selected from the group consisting of glycerol, DMSO, ethanol, acetone, PPG and combinations thereof.

41. The method according to claim 1 or 2, wherein the recovery of the desired activity of the biological material after sterilization by irradiation is at least about 50% of the pre-irradiation value.

42. A method for prophylaxis or treatment of a condition or disease in a mammal comprising administering to a mammal in need thereof an effective amount of a biological material made according to a method of claim 1 or 2.

43. The method according to claim 4, wherein said biological material is tissue and said at least one stabilizer is introduced into said tissue by a method comprising:

(a) soaking said tissue in a solution containing said at least one stabilizer;

(b) applying a gas containing said at least one stabilizer to said tissue;

(c) injecting said at least one stabilizer into said tissue;

(d) injecting a solution containing said at least one stabilizer into said tissue;

(e) placing said tissue under reduced pressure and contacting said tissue with a gas or solution containing said at least one stabilizer; and

(f) dehydrating said tissue and rehydrating said tissue with a solution containing said at least one stabilizer.

44. A biological material made according to a method of claim 1 or 2.

45. The biological material according to claim 44, wherein said biological material is selected from the group consisting of dextrose, urokinase, thrombin, purified protein fraction, blood, blood cells, alpha-1 proteinase inhibitor, digestive enzymes, blood proteins and tissue.

46. The biological material according to claim 45, wherein said tissue is selected from the group consisting of tendons, nerves, bone, teeth, bone marrow, skin grafts, cartilage, corneas, arteries, veins, heart valves, ligaments and demineralized bone matrix.

47. The biological material according to claim 45, wherein said digestive enzymes are selected from the group consisting of galactosidases and sulfatases.

48. The biological material according to claim 45, wherein said blood proteins are selected from the group consisting of albumin, Factor VIII, Factor VII, Factor IV, fibrinogen, monoclonal immunoglobulins and polyclonal immunoglobulins.

49. The biological material according to claim 44, wherein said biological material is milk.

50. The biological material according to claim 44, wherein said biological material is serum or plasma.

2. A method for sterilizing a biological material that is sensitive to radiation, said method comprising;

(a) determining the maximum acceptable temperature (T_max) for said biological material during irradiation; and

(b) irradiating said biological material with radiation for a time effective to sterilize said biological material at a rate effective to sterilize said biological material, wherein said irradiating is performed under conditions whereby the temperature of said biological material increases during said irradiating from an initial temperature (T_i) to a final temperature (T_f), wherein said initial temperature (T_i) is no greater than about T_max−

T_obs, wherein T_obs is determined by a process comprising irradiating a sample of said biological material or a suitable substitute therefore with said radiation under the conditions under which said biological material is to be irradiated in step (b) while measuring the increase in temperature of said sample.