Rapid cryobaric sterilization and vaccine preparation

• US 6,270,723 B1
• Filed: 10/02/1998
• Issued: 08/07/2001
• Est. Priority Date: 06/15/1998
• Status: Expired due to Term

• Current Assignee: Pressure BioSciences, Inc.
• Original Assignee: BBI Bioseq Incorporated
• Inventors: Bradley, David W., Laugharn, James A. Jr., Hess, Robert A.
• Primary Examiner(s): MCKANE, ELIZABETH L
• Application Number:
  US09/165,829
• Time in Patent Office: 1,040 Days
• Field of Search: 422/39, 422/33, 422/40, 422/1, 435/236, 424/184.1, 206/367, 436/543
• US Class Current: 422/39
• CPC Class Codes: A61L 2/0011 using physical methods
• CPC Class Codes: A61L 2/02 using physical phenomena
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention is based on the discovery that biological and non-biological materials can be sterilized, decontaminated, or disinfected by repeatedly cycling between relatively high and low pressures. Pressure cycling can be carried out at low, ambient, or elevated temperatures (e.g., from about −20° C. to about 95° C.). New methods based on this discovery can have applications in, for example, the preparation of vaccines, the sterilization of blood plasma or serum, the decontamination of military devices, and the disinfection of medical equipment. The new methods can also be incorporated into production processes or research procedures.

Claims

1. A method for sterilizing a material comprising at least one desired protein, the method comprising:

providing said material and, while maintaining the temperature of the material at 4°

C. or lower, increasing the pressure to an elevated pressure sufficient to sterilize the material but insufficient to irreversibly denatured the desired protein, thereby providing a sterilized material.

2. The method of claim 1, wherein the initial pressure is about 1 atm.

3. The method of claim 1, wherein the initial temperature is between −

40°

C. and 0°

C.

4. The method of claim 3, wherein the initial temperature is between −

20°

C. and −

5°

C.

5. The method of claim 1, wherein the elevated pressure is in the range of about 5,000 psi to about 50,000 psi.

6. The method of claim 1, further comprising repeatedly cycling the pressure between the elevated pressure and the initial pressure.

7. The method of claim 6, wherein the initial temperature is in the range from about −

40°

C. and 4°

C.

8. The method of claim 1, further comprising cooling the material to a subzero temperature.

9. The method of claim 8, wherein the cooling step is carried out prior to the pressure increasing step.

10. The method of claim 1, wherein the material being sterilized is selected from the group consisting of a biological sample, blood plasma, serum, living tissue, medical or military equipment, a foodstuff, a pharmaceutical preparation, and a vaccine.

11. The method of claim 1, wherein the material being sterilized is initially contaminated with at least one of a bacterium, a virus, a fungus, a protist, a nucleic acid, and a protein.

12. The method of claim 1, wherein a protein stabilizing agent is added prior to sterilization.

13. The method of claim 12, wherein the protein stabilizing agent is selected from the group consisting of sugars, glycerol, a hydrophilic polymer, a cyclodextrin, a caprylate, acetyl tryptophanoate, polyethylene glycol, anti-oxidant, and a protein specific ligand.

14. The method of claim 1, wherein the material to be sterilized is provided in its final packaging, the packaging being adapted to transmit pressure without rupture.

15. The method of claim 14, wherein the packaging is hermetically sealed in flexible plastic.

16. The method of claim 14, wherein the packaging is a syringe and pressure is transmitted via a plunger.

26. A method of producing a vaccine against a pathogen, the method comprising:

obtaining a suspension of pathogenic cells;

sterilizing the suspension by the method of claim 15; and

adding an adjuvant to the sterilized suspension to produce a vaccine.

27. The method of claim 26, further comprising removing toxins from the suspension after the sterilizing step.

28. The method of claim 26, wherein the material to be sterilized is provided in its final packaging, the packaging being adapted to transmit pressure without rupture.

29. The method of claim 28, wherein the packaging is hermetically sealed in flexible plastic.

30. The method of claim 28, wherein the packaging is a syringe and a pressure is transmitted via a plunger.

32. A material sterilized by the method of claim 1.

17. A method for sterilizing a material comprising at least one desired protein, the method comprising:

providing said material and, while maintaining the temperature of the material below 25°

C., increasing the pressure to an elevated pressure sufficient to sterlize the material but insufficient to irreversibly denature the desired protein, wherein the elevated pressure is in the range of about 10,000 to about 30,000 psi, thereby providing a sterilized material.

18. A method for sterilizing a material comprising at least one desired protein, the method comprising:

providing said material and, while maintaining the temperature of the material below 25°

C., increasing the pressure to an elevated pressure sufficient to sterilize the material but insufficient to irreversibly denature to desired protein, and further comprising cooling the material to a subzero temperature after the pressure increasing step,

thereby providing a sterilized material.

19. A method for sterilizing a material comprising at least one desired protein, the method comprising:

providing said material and, while maintaining the temperature of the material below 25°

C., increasing the pressure to an elevated pressure sufficient to sterilize the material but insufficient to irreversibly denature the desired protein, and further comprising cooling the material to a decreased temperature, wherein the decreased temperature is in the range from about −

40°

C. to about 4°

C.

thereby providing a sterilized material.

20. The method of claim 19, wherein the decreased temperature is in the range from about −

20 to about −

5°

C.

21. A method for sterilizing a material comprising at least one desired protein, the method comprising:

providing said material and, while maintaining the temperature of the material below 25°

C., increasing the pressure to an elevated pressure sufficient to sterilize the material but insufficient to irreversibly denature the desired protein;

warming the material to the initial temperature, and then

decreasing the pressure,

thereby providing a sterilized material.

22. A method for sterilizing a material comprising at least one desired protein, the method comprising:

providing said material and, while maintaining the temperature of the material below 25°

C., increasing the pressure to an elevated pressure sufficient to sterilize the material but insufficient to irreversibly denature the desired protein;

decreasing the pressure; and

then

warming the material to the initial temperature, thereby providing a sterilized material.

23. A method for sterilizing a material comprising at least one desired protein, the method comprising:

providing said material at an initial pressure and temperature;

cooling the material to decreased temperature below 25°

C., then, while maintaining the temperature of the material at the decreased temperature,

increasing the pressure to an elevated pressure sufficient to sterilize the material but insufficient to irreversibly denature the desired protein, and

repeatedly cycling the pressure between the elevated pressure and the initial pressure,

thereby providing a sterilized material.

24. The method of claim 23, wherein said decreased temperature is in the range from about −

40°

C. to about 4°

C.

25. The method of claim 23, wherein the decreased temperature is in the range from about −

20°

C. to about −

5°

C.

31. A method for sterilizing a material, the method comprising:

providing a material at an initial pressure and temperature;

increasing the pressure to an elevated pressure insufficient to irreversibly denature proteins of interest; and

decreasing the pressure to the initial pressure, thereby providing a sterilized material, wherein the material to be sterilized includes a phase-change catalyst.

33. A method for sterilizing a sample that includes at least one desired protein, the method comprising:

providing the sample at an initial pressure;

increasing the pressure to a pressure sufficient to inactivate pathogens;

maintaining the elevated pressure for a period of less than about 5 seconds; and

restoring the initial pressure to provide a sterilized sample and to avoid substantial aggregation of the desired protein.