Methods of using pneumatic bioreactors

• US 8,790,913 B2
• Filed: 09/17/2010
• Issued: 07/29/2014
• Est. Priority Date: 10/26/2005
• Status: Active Grant

• Current Assignee: PBS Biotech, Inc.
• Original Assignee: PBS Biotech, Inc.
• Inventors: Zeikus, J. Gregory
• Primary Examiner(s): SOOHOO, TONY GLEN
• Application Number:
  US12/884,431
• Publication Number:
  US 20110003366A1
• Time in Patent Office: 1,411 Days
• Field of Search: 366/101, 435/302.1, 435/289.1, 435/295.2, 435/296.1, 435/257.1, 47/655
• US Class Current: 435/257.1
• CPC Class Codes: B01F 23/233 using driven stirrers with completely immersed stirring elements
• CPC Class Codes: B01F 23/23351 the gas moving along the axis of rotation
• CPC Class Codes: B01F 23/23362 the gas being introduced under the stirrer
• CPC Class Codes: B01F 27/073 with stirring elements moving with respect to the stirrer shaft, e.g. floating or comprising contracting chambers
• CPC Class Codes: B01F 27/11252 paddle wheels
• CPC Class Codes: B01F 27/55 with stirrers driven by the moving material
• CPC Class Codes: B01F 27/70 with paddles, blades or arms
• CPC Class Codes: B01F 27/91 with propellers
• CPC Class Codes: C12M 23/28 disposable or single use
• CPC Class Codes: C12M 23/56 Floating elements
• CPC Class Codes: C12M 27/00 Means for mixing, agitating or circulating fluids in the vessel by introduction of gas C12M29/06, C12M29/14, mixing in general or mixers per se B01F; mixing in apparatus for chemical or physical processes B01J
• CPC Class Codes: C12M 27/06 with horizontal or inclined stirrer shaft or axis
• CPC Class Codes: C12M 37/00 Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination C12M23/38 takes precedence; filtration in general and filters per se B01D24/00-B01D41/00; autoclaves B01J3/04; treatment of microorganisms with electrical or wave energy C12N13/00
• CPC Class Codes: C12M 41/12 of temperature controlling the temperature of chemical or physical processes B01J19/0013, heating or cooling apparatus for laboratory use B01L7/00
• CPC Class Codes: C12M 41/42 of agitation speed

Abstract

A pneumatic bioreactor includes a vessel containing a fluid to be mixed and at least one mixing device driven by gas pressure. A first embodiment includes a floating impeller that rises and falls in the fluid as gas bubbles carry it upward to the surface where the gas is then vented, permitting the impeller to sink in the fluid. The floating impeller may be tethered to a second impeller with a flexible member and pulley. The mixing speed is controlled with electromagnets in the vessel acting upon magnetic material in the impeller or its guides. In another embodiment, floating pistons mix the fluid, pushing it through a mixing plate with one or more apertures. In a third embodiment, the mixing device is a rotating drum with bubble-catching blades and rotating mixing plates with apertures. The top of the vessel for these mixers may include a closed top and sterile filters.

Claims

1. A method of growing a biological agent within a single use bioreactor, the method comprising the steps of:

providing a containment vessel for holding biological contents, the containment vessel comprising a generally rectangular upper portion in horizontal section and a semi-cylindrical lower portion defining a semi-cylindrical concavity therewithin;

providing a rotatable buoyancy-driven mixing device located within the containment vessel and having a substantially horizontal axis of rotation, the mixing device having a diameter such that when it rotates it substantially fills the semi-cylindrical concavity;

sterilizing the containment vessel and the mixing device so that at least the interior of the containment vessel and the mixing device are sterile;

providing a culture medium within the containment vessel;

providing nutrients within the containment vessel to support growth of the biological agent within the culture medium;

enclosing the containment vessel such that the culture medium is not exposed to the surrounding environment;

rotating the mixing device about said horizontal axis to mix the contents of the containment vessel by introducing gas into the containment vessel at a location below the mixing device such that at least a portion of the gas rising within the containment vessel is temporarily captured by a portion of the mixing device to cause rotation of the mixing device due to the buoyancy of the captured gas; and

discarding the containment vessel and mixing device after a single use of growing a biological agent.

2. The method of claim 1, further comprising controlling the temperature of the interior of the containment vessel to support growth of the biological agent.

3. The method of claim 1, further comprising controlling the speed of rotation of the mixing device by controlling the rate of introduction of gas into the containment vessel.

4. The method of claim 1, further comprising controlling the direction of rotation of the mixing device by introducing gas into the containment vessel below one side or the opposite side of the mixing device.

5. The method of claim 1, wherein said sterilizing step comprises sterilizing the containment vessel and the mixing device by gamma irradiation.

6. The method of claim 1, further including a rigid housing into which the containment vessel fits, the housing defining a semi-cylindrical lower section and the lower portion of the containment vessel being formed of a material that is non-structural such that the semi-cylindrical lower section of the housing structurally supports the lower portion of the containment vessel.

7. The method of claim 6, wherein a vessel top extends outwardly of containment vessel sides and rests on upper edges of the housing so as to suspend the containment vessel within the housing.

8. The method of claim 6, wherein the containment vessel is primarily formed of a thin wall film that is non-structural.

9. The method of claim 1, further including an impeller defined within an outer wheel on the mixing element configured to generate axial flow of at least a portion of the contents of the containment vessel upon rotation of the mixing device.

10. A method for performing a cell culture growth process comprising:

holding a cell culture medium within a single use sterile vessel;

enclosing the vessel such that the cell culture medium is not exposed to a surrounding environment;

adding nutrients to the contents of the vessel to support cell culture growth;

maintaining the contents of the vessel at a prescribed temperature to support cell culture growth;

introducing gas into the vessel to support cell culture growth;

mixing the contents of the container with a pneumatically-driven mixing element disposed within the vessel so as to be rotatable about a horizontal axis and being constructed and arranged to capture at least a portion of the gas introduced into the vessel and to be rotated by buoyant force of the captured gas to thereby effect mixing of the contents of the vessel, wherein the mixing element has at least one impeller blade located radially inward from an outer edge thereof configured to generate axial flow of at least a portion of the contents of the vessel upon rotation of the mixing element; and

discarding the vessel and mixing element after a single cell culture growth process.

11. The method of claim 10, further comprising controlling the speed of rotation of the mixing element by controlling the rate of introduction of gas into the vessel.

12. The method of claim 10, wherein the gas is introduced at a location below the mixing element.

13. The method of claim 12, further comprising controlling the direction of rotation of the mixing element by introducing gas into the vessel below one side or the opposite side of the mixing element.

14. The method of claim 10, wherein the vessel is sterilized by exposing the vessel and the mixing element to gamma irradiation.

15. The method of claim 10, wherein the mixing element is configured to reduce force imposed on the mountings of the mixing element by generating toroidal flow of at least a portion of the contents of the vessel upon rotation of the mixing element.

16. The method of claim 10, wherein the vessel comprises a generally rectangular upper portion in horizontal section and a semi-cylindrical lower portion defining a semi-cylindrical concavity therewithin, and wherein the mixing element has a diameter such that when it rotates it substantially fills the semi-cylindrical concavity.

17. The method of claim 16, further including a rigid housing into which the vessel fits, the housing defining a semi-cylindrical lower section and the lower portion of the vessel being formed of a material that is non-structural such that the semi-cylindrical lower section of the housing structurally supports the vessel.

18. The method of claim 17, wherein a vessel top extends outwardly of vessel sides and rests on upper edges of the housing so as to suspend the vessel within the housing.

19. The method of claim 17, wherein the vessel is primarily formed of a thin wall film that is non-structural.

20. The method of claim 10, wherein the impeller is defined within an outer wheel on the mixing element.