Creation of shear in a reactor

• US 20050277187A1
• Filed: 06/07/2005
• Published: 12/15/2005
• Est. Priority Date: 06/07/2004
• Status: Abandoned Application

• Current Assignee: BioProcessors Corp.
• Original Assignee: BioProcessors Corp.
• Inventors: Aumond, Bernardo D., Zarur, Andrey J., Johnson, Timothy J., Rodgers, Seth T., Vella, George J., Benoit, Brian O.
• Application Number:
  US11/147,416
• Publication Number:
  US 20050277187A1
• Time in Patent Office: Days
• US Class Current: 435/293.100
• CPC Class Codes: B01F 29/64 with stirring devices moving in relation to the receptacle, e.g. rotating
• CPC Class Codes: B01F 33/25 Mixers with loose mixing elements, e.g. loose balls in a receptacle
• CPC Class Codes: B01F 33/251 using balls as loose mixing element
• CPC Class Codes: B01F 33/252 using bubbles as loose mixing element
• CPC Class Codes: B01F 33/253 using sliders or cylindrical elements as loose mixing element
• CPC Class Codes: B01F 33/30 Micromixers
• CPC Class Codes: B01F 33/35 Mixing after turning the mixing vessel upside down
• CPC Class Codes: B01F 33/452 using independent floating stirring elements
• CPC Class Codes: B01F 33/453 using supported or suspended stirring elements
• CPC Class Codes: B01J 2219/00488 by rotation of the reaction vessels
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502746 characterised by the means for controlling flow resistance, e.g. flow controllers, baffles B01L3/502738 takes precedence
• CPC Class Codes: C12M 23/16 Microfluidic devices; Capillary tubes integrated microfluidic structures B01L3/5027; microreactors B01J19/0093
• CPC Class Codes: C12M 23/24 Gas permeable parts
• CPC Class Codes: C12M 35/04 : Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli

Abstract

An apparatus for performing a biological or biochemical reaction that, in certain embodiments, has the ability to apply shear stress to a component of a liquid sample and includes a biological or biochemical reactor comprising a container having a volume of less than about 2 mL and containing a liquid sample, and a shear-generating element, the shear-generating element being contained within the apparatus and constructed and arranged so that the entire shear-generating element moves along a selected path of motion intersecting a first location within the apparatus and a second location within the apparatus, with or without rotational movement is described. A method of applying shear stress to a component of a liquid sample that includes moving a liquid or gaseous shear-generating element within an apparatus along a selected path of motion to create a reproducible and controllable level of shear stress at a selected location within the liquid sample is also disclosed.

Claims

1. An apparatus for performing a biological or biochemical reaction having the ability to apply shear stress to a component of a liquid sample, comprising:

a biological or biochemical reactor comprising a container having a volume of less than about 2 mL, the container containing a liquid sample; and

a shear-generating element, which does not comprise a surface of a container or a conduit in contact with a liquid, the shear-generating element being contained within the apparatus and constructed and arranged so that the entire shear-generating element moves along a selected path of motion intersecting a first location within the apparatus and a second location within the apparatus, with or without rotational movement.

2. An apparatus as in claim 1, wherein the shear-generating element is contained and movable within the container containing the liquid sample.

3. An apparatus as in claim 1, wherein the container is constructed and arranged to facilitate cell cultivation.

4. An apparatus as in claim 3, wherein the container is constructed and arranged to facilitate mammalian cell cultivation.

5. An apparatus as in claim 1 further comprising:

a first gas permeable, liquid vapor impermeable membrane defining a first wall of the container.

6. An apparatus as in claim 1, wherein the shear-generating element has an average density at least 1% different from the average density of the liquid sample.

7. An apparatus as in claim 1, wherein the shear-generating element has an average density at least 5% different from the average density of the liquid sample.

8. An apparatus as in claim 1, wherein the container has a volume of less than about 1.3 mL.

9. An apparatus as in claim 1, wherein changes in the movement of the shear-generating element that create changes in a level or pattern of shear stress within the liquid sample do not significantly affect gas exchange between the liquid sample and the exterior of the reactor.

10. An apparatus as in claim 1, wherein the shear-generating element is a gas bubble contained within the apparatus.

11. An apparatus as in claim 1, wherein the shear-generating element is a gas bubble contained and movable within the container containing the liquid sample.

12. An apparatus as in claim 1, further comprising a control system configured to control the movement of the shear-generating element to facilitate the creation of a reproducible and controllable level of shear stress at a selected location within the liquid sample.

13. An apparatus as in claim 1, wherein the gas permeable, liquid vapor impermeable membrane has an oxygen permeability of greater than or equal to 0.061 mol O₂/(day·

m2·

atm).

14. An apparatus as in claim 13, wherein the membrane has an oxygen permeability of less than or equal to 0.6 mol O₂/(day·

m2·

atm).

15. An apparatus as in claim 1, wherein a second gas permeable, liquid vapor impermeable membrane defines a second wall of the container.

16. An apparatus as in claim 2, wherein the thickness of the container varies along the selected path of motion of the shear-generating element.

17. An apparatus as in claim 1, further comprising a plurality of containers each having a volume of less than about 2 mL and each containing a liquid sample.

18. An apparatus as in claim 17, wherein the plurality of containers are present on a chip, and the chip is constructed and arranged to enable it to be stably connected in a selected orientation with respect to other, similar chips in the apparatus.

19. An apparatus as in claim 1, wherein the shear-generating element is a solid element.

20. An apparatus as in claim 1, wherein the shear-generating element is a liquid immiscible in the liquid sample.

21. An apparatus as in claim 1, further comprising a rotating apparatus to which the container is attached.

22. An apparatus as in claim 1, wherein the chip further comprises a predetermined gas region in fluid communication with the container, in which the shear-generating element can be positioned when the shear-generating element is not being used to generate shear.

23. An apparatus as in claim 1, further comprising an inlet port, an outlet port, and a self-sealing elastomeric material defining portions of the inlet port and the outlet port.

24. An apparatus as in claim 1, wherein the container is defined by a void in a substrate layer.

25. An apparatus as in claim 5, wherein an adhesive layer binds the gas permeable vapor impermeable membrane to the substrate layer.

26. An apparatus for performing a biological or biochemical reaction having the ability to apply shear stress to a component of a liquid sample, comprising:

a biological or biochemical reactor comprising a container containing a liquid sample; and

a shear-generating element, which does not comprise a surface of a container or a conduit in contact with a liquid, the shear-generating element being contained within the container and constructed and arranged so that the entire shear-generating element moves along a selected path of motion intersecting a first location within the container and a second location within the container, wherein

changes in the movement of the shear-generating element within the container that create changes in a level or pattern of shear stress in the liquid sample do not significantly affect gas exchange between the liquid sample and the exterior of the reactor.

27. An apparatus as in claim 26, further comprising a first gas permeable, liquid vapor impermeable membrane defining a first wall of the container.

28. An apparatus as in claim 27, further comprising an inlet port, an outlet port, and at least one microfluidic channel in fluid communication with the container.

29. An apparatus as in claim 27, wherein the container is constructed and arranged to maintain at least one living mammalian cell.

30. An apparatus as in claim 26, wherein the shear-generating element has an average density at least 1% different from the average density of the liquid sample.

31. An apparatus as in claim 26, wherein the container has a volume of less than about 5 mL.

32. An apparatus as in claim 26, wherein the shear-generating element is a gas bubble.

33. An apparatus as in claim 26, further comprising a control system configured to control the movement of the shear-generating element to facilitate the creation of the reproducible and controllable level of shear stress at a selected location within the liquid sample.

34. An apparatus as in claim 26, wherein the shear-generating element is a solid element attached to the container.

35. An apparatus as in claim 34, wherein the shear-generating element is slidingly attached to the container.

36. An apparatus as in claim 26, wherein the shear-generating element is a liquid immiscible in the liquid sample.

37. An apparatus for performing a biological or biochemical reaction having the ability to apply shear stress to a component of a liquid sample, comprising:

a biological or biochemical reactor comprising a container, the container containing a liquid sample;

a shear-generating element within the apparatus that is movable within the apparatus upon inversion of the apparatus; and

a control system configured to control movement of the shear-generating element to facilitate creation of a reproducible and controllable level of shear stress at a selected location within the liquid sample.

38. An apparatus as in claim 37, wherein the container has a volume of less than 5 mL.

39. An apparatus as in claim 37, wherein the container has a volume of greater than 0.01 mL and less than 3 mL.

40. An apparatus as in claim 37, wherein the container has a volume of greater than 0.5 mL and less than 3 mL.

41. An apparatus as in claim 37, further comprising a first gas permeable, liquid vapor impermeable membrane defining a first wall of the container.

42. An apparatus as in claim 37, wherein the shear-generating element has an average density at least 1% different from the average density of the liquid sample.

43. An apparatus as in claim 37, wherein the shear-generating element is a gas bubble.

44. An apparatus as in claim 37, wherein the shear-generating element is a gas bubble contained and movable within the container containing the liquid sample.

45. A method of applying shear stress to a biological or biochemical component of a liquid sample contained within a container, comprising:

moving and/or controlling movement of a shear-generating element within a container containing a liquid sample, the movement of the shear-generating element occurring upon inversion of the container, wherein the movement applies a reproducible and controllable level of shear stress to a biological or biochemical component at a selected location within the liquid sample.

46. A method as in claim 45, wherein the container is capable of maintaining at least one living cell.

47. A method as in claim 45, wherein the container is capable of maintaining at least one living mammalian cell.

48. A method as in claim 45, a gas permeable, liquid vapor impermeable membrane defines a first wall of the container.

49. A method as in claim 45, wherein the movement applies a preselected level of shear stress at the selected location within the liquid sample.

50. A method as in claim 49, further comprising an additional and separate act of moving and/or controlling movement of the shear-generating element within the container containing a liquid sample, the movement of the shear-generating element occurring upon inversion of the container, wherein the additional and separate movement applies a different preselected level of shear stress at the selected location within the liquid sample.

51. A method as in claim 45, wherein the shear-generating element is a gas bubble.

52. A method as in claim 45, wherein the shear-generating element is a liquid immiscible in the liquid sample.

53. A method as in claim 45, wherein the container is less than approximately 5 mL.

54. A method as in claim 45, wherein the container is less than approximately 1 mL.

55. A method as in claim 45, wherein the container is less than approximately 0.01 mL.

56. A method as in claim 45, wherein moving and/or controlling movement of the shear-generating element comprises rotating the container.

57. A method as in claim 56, wherein rotating the container comprises rotating the container about an axis that is external to the container.

58. A method as in claim 56, further comprising rotating the container using discontinuous rotation rates.

59. An apparatus for performing a biological or biochemical reaction having the ability to apply shear stress to a component of a liquid sample, comprising:

a biological or biochemical reactor comprising a container configured to contain a liquid sample, a surface of the container comprising a membrane having an oxygen permeability of greater than or equal to 0.061 mol O₂/(day·

m2·

atm); and

a shear-generating element contained within the container and constructed and arranged so that the entire shear-generating element moves along a selected path of motion intersecting a first location within the container and a second location within the container during operation when the container contains the liquid sample.

60. An apparatus as in claim 59, wherein the container has a volume of less than 5 mL.

61. An apparatus as in claim 60, wherein the container contains the liquid sample and wherein the shear-generating element is a gas bubble.

62. An apparatus as in claim 59, wherein the shear-generating element is a solid element.

63. A method as in claim 59, wherein the container contains the liquid sample and wherein the shear-generating element is a liquid immiscible in the liquid sample.

64. An apparatus as in claim 59, wherein the membrane has an oxygen permeability of less than or equal to 69.7 mol O₂/(day·

m2·

atm).

65. An apparatus as in claim 59, wherein the membrane has an oxygen permeability of less than or equal to 140 mol O₂/(day·

m2·

atm).

66. An apparatus as in claim 59, further comprising a control system configured to control the movement of the shear-generating element relative to the container so as to create a reproducible and controllable level of shear stress at a selected location within the liquid sample.

67. A method of applying shear stress to a biological or biochemical component of a liquid sample comprising:

moving an entire shear-generating element, freely suspended within an apparatus, along a selected path of motion intersecting a first location within the apparatus and a second location within the apparatus to apply a reproducible and controllable level of shear stress to a biological or biochemical component at a selected location within a liquid sample, wherein the shear-generating element is either a gas or a liquid.

68. A method as in claim 67, wherein the liquid sample is contained within a reaction site container that is capable of maintaining at least one living cell.

69. A method as in claim 68, wherein the at least one living cell is a mammalian cell.

70. A method as in claim 67, further comprising a gas permeable, liquid vapor impermeable membrane defining a first wall of the container.

71. A method as in claim 67, wherein the moving the shear-generating element a preselected level of shear stress at the selected location within the liquid sample.

72. A method as in claim 67, wherein the shear-generating element is a gas bubble.

73. A method as in claim 67, wherein the shear-generating element is a liquid immiscible in the liquid sample.

74. A method as in claim 67, wherein moving the shear-generating element comprises changing the orientation of the apparatus relative to the direction of gravity.

75. A method as in claim 67, wherein moving the shear-generating element comprises applying a magnetic field to the shear-generating element.

76. A method as in claim 67, wherein moving the shear-generating element comprises applying an electrical field to the shear-generating element.

77. A method as in claim 67, wherein the shear-generating element is freely suspended within a container having a volume of less than about 2 mL.

78. A method as in claim 67, further comprising:

receiving feedback of at least one measurement from the liquid sample; and

adjusting at least one control parameter of the apparatus in response to the measurement.

79. A method as in claim 67, further comprising:

receiving feedback of at least one measurement from the liquid sample; and

operating simulation software to determine at least one parameter value for controlling the apparatus.

80. An apparatus for performing a biological or biochemical reaction having the ability to apply shear stress to a component of a liquid sample, comprising:

a biological or biochemical reactor comprising a container having a volume of less than about 2 mL and containing a liquid sample; and

a shear-generating element, which does not comprise a surface of a container or a conduit in contact with a liquid, the shear-generating element being contained within the container and constructed and arranged for pivoting movement within the container, the pivoting movement creating a reproducible and controllable level of shear stress at a selected location within the liquid sample.

81. An apparatus as in claim 80, wherein the shear-generating element is a member having an end that is pivotally attached to an interior surface of the container.