CONTINUOUS PERFUSION BIOREACTOR SYSTEM

• US 20090035856A1
• Filed: 05/20/2008
• Published: 02/05/2009
• Est. Priority Date: 07/30/2007
• Status: Active Grant

• Current Assignee: Global Life Sciences Solutions USA LLC (Danaher Corporation)
• Original Assignee: Xcellerex, Inc. (General Electric Company)
• Inventors: Mardirosian, Dan, Guertin, Patrick, Fisher, Michael, Hodge, Geoffrey L., Galliher, Parrish M.
• Granted Patent: US 9,109,193 B2
• Time in Patent Office: Days
• US Class Current: 435/383
• CPC Class Codes: B01D 29/13 Supported filter elements
• CPC Class Codes: C12M 23/14 Bags
• CPC Class Codes: C12M 23/26 flexible flexible containers for laboratory use B01L3/505
• CPC Class Codes: C12M 23/48 Holding appliances; Racks; Supports holding devices for laboratory apparatus B01L9/00
• CPC Class Codes: C12M 27/02 Stirrer or mobile mixing elements
• CPC Class Codes: C12M 27/20 Baffles; Ribs; Ribbons; Auger vanes
• CPC Class Codes: C12M 29/00 Means for introduction, extraction or recirculation of materials, e.g. pumps pumps per se F04B
• CPC Class Codes: C12M 29/10 Perfusion
• CPC Class Codes: C12M 29/18 External loop; Means for reintroduction of fermented biomass or liquid percolate loop type reactors for chemical or physical processes B01J19/2435
• CPC Class Codes: C12M 47/10 Separation or concentration of fermentation products bioreactors combined with means for distillation or extraction of liquid fuel C12M43/02

Abstract

Systems and methods for containing and manipulating liquids, including vessels and unit operations or components of cell culture, cell containment, bioreactor, and/or pharmaceutical manufacturing systems, are provided. In certain embodiments, such vessels and unit operations are directed to continuous perfusion reactor or bioreactor systems and may include one or more disposable components. For instance, in one aspect, a system includes an apparatus in the form of a bioreactor for harvesting cells which produce one or more products. The apparatus may include a disposable, collapsible bag adapted for containing a liquid, the collapsible bag in fluid communication with a liquid-solids (e.g., cell) separation device. For example, an outlet of the collapsible bag may be connected to an inlet of the separation device, and an outlet of the separation device may be connected to an inlet of the collapsible bag for recycle. Accordingly, after separating the cells from the liquid in the separation device, the cells can be flowed back into the collapsible bag where they can be re-harvested. Meanwhile, product contained in the liquid can be collected in a separate container. The efficiency of product formation in such a system may be enhanced by using mixing systems described herein.

Claims

1. A system, comprising:

a first apparatus, comprising;

a first collapsible bag adapted for containing a liquid, the first collapsible bag including a first inlet, a first outlet, and a base plate that is attached to the first collapsible bag;

an impeller associated with the base plate;

a first reusable support structure adapted for surrounding and supporting the first collapsible bag; and

a second apparatus in fluid communication with the first apparatus, the second apparatus including a second inlet and a second outlet, wherein the second apparatus is disposed external to the first collapsible bag or disposed inside of the first collapsible bag,

with the proviso that, if the second apparatus is disposed external to the first collapsible bag, the first outlet of the first collapsible bag is in fluid communication with the second inlet of the second apparatus, and the second outlet of the second apparatus is in fluid communication with the first inlet of the first collapsible bag; and

if the second apparatus is disposed inside of the first collapsible bag, the first outlet of the first collapsible bag is in fluid communication with the second outlet of the second apparatus, and the second inlet of the second apparatus is in direct fluid communication with the liquid inside the first collapsible bag.

2. The system of claim 1, wherein the second apparatus is disposed external to the first apparatus and is configured to receive a first liquid comprising a first concentration of a component from the first apparatus and to deliver a second liquid comprising a second concentration of the component to the first apparatus, wherein the first and second concentrations are different.

3. The system of claim 2, wherein the component comprises a substance chosen from a viable or non-viable organism, a precipitate, a cell, a polymeric porous or nonporous sphere, a solid sphere, a gelatinous particle, a microbead, a microdisk, a cross-linked bead, a biochemical molecular entity, and combinations thereof.

4. The system of claim 1, wherein the the second apparatus is disposed inside of the first collapsible bag and is configured to receive through the second inlet of the second apparatus a first liquid comprising a first concentration of a component directly from the liquid inside the first collapsible bag and to deliver a second concentration of the component to the first apparatus, wherein the first and second concentrations are different.

5. The system of claim 4, wherein the component comprises a substance chosen from a viable organism, a non-viable organism, a precipitate, a cell, a polymeric porous or nonporous sphere, a solid sphere, a gelatinous particle, a microbead, a microdisk, a cross-linked bead, a biochemical molecular entity, and combinations thereof.

8. The system of claim 2, wherein the component comprises a cell growing on a solid object chosen from a solid sphere, a fiber, a gelatinous particle, a polymeric particle, a non-porous particle, a porous particle, and combinations thereof.

9. The system of claim 1, wherein the second apparatus further comprises a second collapsible bag, and a second reusable support structure adapted for surrounding and supporting the second collapsible bag.

11. The system of claim 1, wherein the fluid communication of the second apparatus with the first apparatus during use is continuous or periodic.

14. The system of claim 1, wherein a shaft is associated with the base plate, which is irreversibly attached to the first collapsible bag.

15. The system of claim 1, wherein the impeller is chosen from a magnetically-driven impeller and a direct-shaft driven impeller.

16. The system claim 1, wherein the impeller comprises an impeller support affixed to a side of the first collapsible bag at a lower portion thereof, such that the impeller is off-center from the center of the bottom of the bag.

17. The system of claim 1, wherein the first apparatus forms at least part of a bioreactor system or at least part of a pharmaceutical manufacturing system.

19. The system of claim 1, wherein the first collapsible bag has a volume of between about 1 L and about 1,000 L.

21. The system of claim 1, wherein the second apparatus forms at least part of a separation device.

22. The system of claim 21, wherein the separation device comprises a separation device chosen from a centrifuge, a settling device, a hollow fiber filter, a flat membrane filter, a spiral wound filter, a spin filter, depth filters, tangential flow filters, microporous flow filters, ultrafiltration filters, a plate and frame filtration device, and combinations thereof.

28. The system of claim 1, further comprising a liquid-solids separation device positioned on a surface of or in an internal portion of the first collapsible bag.

31. The system of claim 28, wherein the separation device comprises a device chosen from a microporous frit, an ultrafiltration membrane, a spin filter, a centrifuge, and a settling device.

32. The system of claim 1, wherein the second apparatus forms at least part of a device chosen from a media or buffer storage tank, a mixing system, and a batch, periodic or continuous harvesting system.

36. The system of claim 1, wherein the first support structure comprises one or more baffles mounted either internally or externally as part of the collapsible bag or as part of the external support structure.

37. The system of claim 1, wherein at least one of the first apparatus and the second apparatus is associated with its own ventilation system, cooling system, feedback control system, component that can allow one or both of the apparatuses to be operated independently of one another, and combinations thereof.

38. The system of claim 1, wherein the first collapsible bag comprises one or more internal partitions.

39. The system of claim 1,

wherein at least one of the first apparatus and the second apparatus is in the form of a portable module, and

wherein upon fluid communication between the first and second apparatuses, each apparatus can be moved relative to the other without breaking the connection therebetween.

58. A method of recovering and recycling solutes or cells from a bioreactor comprising the first collapsible bag of the system of claim 21, while maintaining a substantially constant volume of liquid in the bioreactor, the method comprising:

allowing a cell culture suspension in a liquid medium in the first collapsible bag to grow to a first concentration of solutes or cells;

continuously allowing a portion of the liquid medium comprising the solute or cell culture suspension having the first concentration of solutes or cells to flow out of the first outlet of the first collapsible bag and into the second inlet of the second external apparatus comprising the separation device or through the second internal separation device to the outlet of the first apparatus;

allowing the separation device of the second apparatus to separate out at least a portion of the solutes or cells from the liquid medium flowing into the second apparatus from the first collapsible bag, while simultaneously causing a liquid to flow from the second apparatus out of the second outlet of the second apparatus and into the first inlet of the first collapsible bag; and

recycling at least a portion of the solutes or cells separated out by the second apparatus by flowing some or all of the separated solutes or cells back into the collapsible bag, or directly back into the fluid within the first apparatus,

while continuously maintaining a substantially constant volume within the collapsible bag during the outflowing, inflowing, and recycling steps, thereby recovering and recycling solutes or cells from the bioreactor while maintaining a substantially constant volume of liquid in the bioreactor or pharmaceutical device.

59. The system of claim 1, wherein at least one sparger is connected to the first collapsible bag.

60. The system of claim 1, wherein at least one sparger is associated with the base plate.

61. The system of claim 1, wherein at least one sparger is incorporated into an impeller support.

6-7. -7. (canceled)

10. (canceled)

12-13. -13. (canceled)

18. (canceled)

20. (canceled)

23-27. -27. (canceled)

29-30. -30. (canceled)

33-35. -35. (canceled)

40-49. -49. (canceled)

50. A method, comprising:

transferring a first liquid comprising a plurality of substances chosen from biochemical solutes, particulates, solid objects, and combinations thereof from a first collapsible bag to an apparatus including a liquid-solids separation device,

wherein the apparatus including the liquid-solids separation device is external or internal to the collapsible bag, and wherein the first collapsible bag is supported by a first reusable support structure adapted for surrounding and supporting the first collapsible bag;

separating at least a portion of the plurality of the biochemical solutes, particulates, solid objects, or a combination thereof from the first liquid in the apparatus; and

transferring a second liquid from the apparatus to the first collapsible bag, wherein the first and second liquids have different concentrations of the biochemical solutes, particulates, solid objects, or a combination thereof.

51. The method of claim 50, wherein at least a portion of the plurality of biochemical solutes, particulates, solid objects, or a combination thereof separated from the first liquid in the apparatus are chosen from biochemicals, cells, solids with or without cells on the surface, semi-rigid spheres, polymeric spheres, precipitates, and combinations thereof, and

the second liquid transferred from the apparatus to the first collapsible bag comprises a greater concentration of the biochemical solutes, particulates, solid objects, or a combination thereof than the concentration of biochemical solutes, particulates, solid objects, or a combination thereof in the first liquid, thereby flowing some or all of the separated biochemical solutes, particulates, solid objects, or a combination thereof back into the first collapsible bag.

52-57. -57. (canceled)