Devices and methods for magnetic enrichment of cells and other particles

• US 20060223178A1
• Filed: 12/29/2005
• Published: 10/05/2006
• Est. Priority Date: 04/05/2005
• Status: Abandoned Application

• Current Assignee: Gpb Scientific, LLC, Mehmet Toner
• Original Assignee: Mehmet Toner
• Inventors: Schmidt, Martin, Toner, Mehmet, Kapur, Ravi, Huang, Lotien Richard, Barber, Tom
• Application Number:
  US11/323,971
• Publication Number:
  US 20060223178A1
• Time in Patent Office: Days
• US Class Current: 435/325
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/043 magnetic forces
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/086 using baffles or other fixed flow obstructions
• 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: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: B01L 3/502776 specially adapted for focusing or laminating flows
• CPC Class Codes: B03C 1/30 Combinations with other devices, not otherwise provided for
• CPC Class Codes: B03C 1/32 acting on the medium containing the substance being separated, e.g. magneto-gravimetric-, magnetohydrostatic-, or magnetohydrodynamic separation
• CPC Class Codes: B03C 2201/18 Magnetic separation whereby the particles are suspended in a liquid
• CPC Class Codes: C12M 35/06 Magnetic means C12M35/02 takes precedence
• CPC Class Codes: G01N 33/5044 involving specific cell types

Abstract

The invention features devices and methods for the enrichment of cells and other desired analytes by employing a magnetic field, alone or in conjunction with size-based separation. The devices and methods may be advantageously employed to enrich for rare cells, e.g., fetal cells or epithelial cells, present in a sample, e.g., maternal blood.

Claims

1. A device for producing a sample enriched in a first cell or component thereof relative to a second component, said device comprising:

(a) a channel through which said first cell or component flows; and

(b) a magnet that produces a magnetic field of between 0.05 and 5.0 Tesla and a magnetic field gradient of between 100 Tesla/m and 1,000,000 Tesla/m in said channel.

2. The device of claim 1, wherein said first cell or component is retained in said channel and said second component is not retained in said channel.

3. The device of claim 1, wherein said first cell or component is not retained in said channel and said second component is retained in said channel.

4. The device of claim 1, wherein said channel comprises first and second outlets, and said first cell or component thereof is directed into said first outlet, while said second component is directed into said second outlet.

5. The device of claim 1, further comprising an analytical module that enriches said first cell or component based on size, shape, deformability, or affinity.

6. The device of claim 5, wherein said analytical module comprises a first channel comprising a structure that deterministically deflects particles having a hydrodynamic size above a critical size in a direction not parallel to the average direction of flow in said structure, wherein said particles are said first cell or component or said second component.

7. The device of claim 1, further comprising a reagent capable of altering a magnetic property of said first cell or component or second component of said sample.

8. The device of claim 7, wherein said reagent alters the magnetic properties of a protein present in said first cell or component or said second component.

9. The device of claim 8, wherein said protein comprises iron.

10. The device of claim 9, wherein said protein is fetal hemoglobin, adult hemoglobin, methemoglobin, myoglobin, or a cytochrome.

11. The device of claim 8, wherein said reagent comprises sodium nitrite, carbon dioxide, oxygen, carbon monoxide, or nitrogen.

12. The device of claim 1, wherein said first cell is a blood cell.

13. The device of claim 1, wherein said first cell is a nucleated cell.

14. The device of claim 1, wherein said first cell is an enucleated cell.

15. The device of claim 1, wherein said blood cell is an adult nucleated red blood cell.

16. The device of claim 1, wherein said blood cell is a fetal nucleated red blood cell.

17. The device of claim 16, wherein said fetal nucleated red blood cell is from a fetus of less than 10 weeks of age.

18. The device of claim 1, wherein said first cell is mammalian, avian, reptilian, or amphibian.

19. The device of claim 1, wherein said component of said first cell is selected from the group consisting of nuclei, peri-nuclear compartments, nuclear membranes, mitochondria, chloroplasts, or cell membranes, lipids, polysaccharides, proteins, nucleic acids, viral particles, or ribosomes.

20. The device of claim 7, wherein said reagent causes expression or overexpression of a protein that is magnetic in said first cell or component or said second component.

21. The device of claim 20, wherein said reagent is capable of transfecting said first cell or said second component with a magnetically responsive protein.

22. The device of claim 7, wherein said reagent comprises a magnetic particle that binds to or is incorporated into said first cell or component or said second component.

23. The device of claim 1, further comprising a pump capable of producing a flow rate of greater than 50,000 cells or components thereof flowing into said channel per second.

24. The device of claim 1, wherein at least 90% of said first cell or component is retained in said device and at least 90% of said second component is not retained in said device.

25. A method for producing a sample enriched in a first cell or component thereof relative to a second component, said method comprising the steps of:

(a) introducing a sample comprising said first cell or component into the device of claim 1;

(b) allowing the passage of said first cell or component or said second component in said sample relative to the other to be altered based on a magnetic property, thereby producing said sample enriched in said first cell or component.

26. The method of claim 25, wherein said sample introduced into said device in step (a) is enriched for said first cell or component relative to a third component.

27. The method of claim 26, wherein, prior to step (a), said sample is contacted with an analytical module that enriches said first cell or component relative to said third component based on size, shape, deformability, or affinity.

28. The method of claim 27, wherein said analytical module comprises a first channel comprising a structure that deterministically deflects particles having a hydrodynamic size above a critical size in a direction not parallel to the average direction of flow in said structure, wherein said particles are said first cell or component or are said third component of said sample.

29. The method of claim 25, wherein said sample enriched in said first cell or component retains at least 70% of said first cells or components present in said sample.

30. The method of claim 25, wherein said sample enriched in said first cell or component is enriched by a factor of 100.

31. The method of claim 25, further comprising the step, prior to step (b), of contacting said sample with a reagent capable of altering a magnetic property of said first cell or component or second component.

32. The method of claim 31, wherein said reagent alters the magnetic properties of a protein present in said first cell or component or said second component.

33. The method of claim 32, wherein said protein is fetal hemoglobin, adult hemoglobin, methemoglobin, myoglobin, or a cytochrome.

34. The method of claim 32, wherein said reagent comprises sodium nitrite, carbon dioxide, or nitrogen.

35. The method of claim 25, wherein said first cell is a blood cell.

36. The method of claim 25, wherein said first cell is a nucleated cell.

37. The method of claim 25, wherein said first cell is an enucleated cell.

38. The method of claim 35, wherein said blood cell is an adult nucleated red blood cell.

39. The method of claim 35, wherein said blood cell is a fetal nucleated red blood cell.

40. The method of claim 39, wherein said fetal nucleated red blood cell is from a fetus of less than 10 weeks of age.

41. The method of claim 25, wherein said first cell is mammalian, avian, reptilian, or amphibian.

42. The method of claim 25, wherein said component of said first cell is nuclei, peri-nuclear compartments, nuclear membranes, mitochondria, chloroplasts, or cell membranes, lipids, polysaccharides, proteins, nucleic acids, viral particles, or ribosomes.

43. The method of claim 31, wherein said reagent causes expression or overexpression of a protein that is magnetic in said first cell or component or said second component.

44. The method of claim 31, wherein said reagent comprises a magnetic particle that binds to or is incorporated into said first cell or component or said second component.

45. The method of claim 25, wherein said sample enriched in said first cell or component comprises at least 90% of said first cell or component in said sample introduced in step (a) and less than 10% of said second component in said sample introduced in step (a).

46. The method of claim 25, wherein greater than 50,000 cells or components thereof flow into said channel per second.

86. The method of claim 69, 1 wherein said first enrichment step comprises a plurality of hydrodynamic size-based enrichment steps that occur in series to one another.

47. A method of producing a sample enriched in a first cell or component thereof relative to a second component, said method comprising the steps of:

(a) contacting a sample comprising said first cell or component with a reagent that alters the magnetic properties of a protein expressed in said first cell or component or said second component of said sample to produce an altered sample;

(b) contacting said altered sample with a channel having a magnet positioned relative to said channel and producing a magnetic field and magnetic field gradient capable of altering the passage of said first cell or component or said second component relative to the other, thereby producing said sample enriched in said first cell or component.

48. The method of claim 47, wherein, prior to or after step (a), said sample comprising said first cell or component is enriched for said first cell or component relative to a third component.

49. The method of claim 48, wherein, prior to step (a), said sample is contacted with an analytical module that enriches said first cell or component relative to said third component based on size, shape, deformability, or affinity.

50. The method of claim 49, wherein said analytical module comprises a first channel comprising a structure that deterministically deflects particles having a hydrodynamic size above a critical size in a direction not parallel to the average direction of flow in said structure, wherein said particles are said first cell or component or are said third component of said sample.

51. The method of claim 47 wherein said sample enriched in said first cell or component retains at least 70% of said first cells or components present in said sample.

52. The method of claim 47, wherein said sample enriched in said first cell or component is enriched by a factor of 100.

53. The method of claim 47, wherein said reagent alters the magnetic properties of a protein present in said first cell or component or said second component.

54. The method of claim 53, wherein said protein is fetal hemoglobin, adult hemoglobin, methemoglobin, myoglobin, or a cytochrome.

55. The method of claim 53, wherein said reagent comprises sodium nitrite, carbon dioxide, or nitrogen.

56. The method of claim 47, wherein said first cell is a blood cell.

57. The method of claim 47, wherein said first cell is a nucleated cell.

58. The method of claim 47 wherein said first cell is an enucleated cell.

59. The method of claim 56, wherein said blood cell is an adult nucleated red blood cell.

60. The method of claim 56, wherein said blood cell is a fetal nucleated red blood cell.

61. The method of claim 60, wherein said fetal nucleated red blood cell is from a fetus of less than 10 weeks of age.

62. The method of claim 47, wherein said first cell is mammalian, avian, reptilian, or amphibian.

63. The method of claim 47, wherein said component of said first cell is nuclei, peri-nuclear compartments, nuclear membranes, mitochondria, chloroplasts, or cell membranes, lipids, polysaccharides, proteins, nucleic acids, viral particles, or ribosomes.

64. The method of claim 47, wherein said reagent causes expression or overexpression of a protein that is magnetic in said first cell or component or said second component.

65. The method of claim 47, wherein said reagent comprises a magnetic particle that binds to or is incorporated into said first cell or component or said second component.

66. The method of claim 47, wherein said sample enriched in said first cell or component comprises at least 90% of said first cell or component in said sample contacted in step (a) and less than 10% of said second component in said sample contacted in step (a).

67. The method of claim 47, wherein a magnet produces a magnetic field of between 0.05 and 5.0 Tesla and a magnetic field gradient of between 100 Tesla/m and 1,000,000 Tesla/m in said channel.

68. The method of claim 47, wherein greater than 50,000 cells or components thereof flow into said channel per second.

69. A method for enriching a first analyte from a fluid sample containing said first analyte relative to second and third analytes in said sample, said method comprising:

(a) performing a first enrichment step to enrich said first analyte from said fluid sample based on hydrodynamic size using a plurality of obstacles that direct said first analyte in a first direction and said second analyte in a second direction, and

(b) performing a second enrichment step to enrich said first analyte from said fluid sample based on an intrinsic or extrinsic magnetic property of said first or third analyte.

70. The method of claim 69, wherein said fluid sample is a blood sample.

71. The method of claim 69, wherein said fluid sample is a maternal blood sample.

72. The method of claim 69, wherein said one or more analytes are red blood cells.

73. The method of claim 69, wherein said one or more analytes are fetal red blood cells.

74. The method of claim 69, wherein each of said one or more analytes comprises fetal hemoglobin, adult hemoglobin, methemoglobin, myoglobin, or a cytochrome.

75. The method of claim 69, wherein said second enrichment step comprises applying a magnetic field to the product of said first enrichment step.

76. The method of claim 75, wherein said magnetic field attracts said first or third analyte.

77. The method of claim 75, wherein said magnetic field repulses said first or third analyte.

78. The method of claim 75, wherein said magnetic field alters the passage of said first analyte relative to said third analyte

79. The method of claim 75, wherein said magnetic field is between 0.5 and 5.0 Tesla.

80. The method of claim 75, wherein said second enrichment step further comprises applying a magnetic field gradient of between 100 Tesla/m and 1,000,000 Tesla/m.

81. The method of claim 69, further comprising the step of deoxygenating said first enrichment product.

82. The method of claim 81, wherein said deoxygenating step comprises contacting the product of said first enrichment step with CO, CO₂, N₂, or NaNO₂.

83. The method of claim 69, further comprising the step of paramagnetizing said first or third analyte.

84. The method of claim 69, further comprising the step of diamagnetizing said first or third analyte.

85. The method of claim 69, wherein said first enrichment step and said second enrichment step occur in series.

87. The method of claim 69, wherein said first enrichment step comprises a plurality of hydrodynamic size-based enrichment steps that occur in parallel to one another.

88. The method of claim 69, wherein said second enrichment step comprises a plurality of enrichment steps that occur in parallel to one another.

89. The method of claim 69, wherein said first enrichment step occurs during sample flow through.

90. The method of claim 69, wherein said second enrichment step occurs during sample flow through.

91. The method of claim 69, wherein said second enrichment step is based on an intrinsic magnetic property.

92. The method of claim 69, wherein said second enrichment step is based on an extrinsic magnetic property.

93. The method of claim 69, wherein greater than 50,000 analytes are subjected to enrichment per second.

94. A system comprising a first module comprising (a) an array of obstacles that selectively directs one or more first analytes having a hydrodynamic size greater than a critical size in a first direction towards a first outlet and one or more second analytes having a hydrodynamic size smaller than said critical size in a second direction towards a second outlet;

(b) a second module comprising a channel for receiving said one or more first analytes from said first outlet; and

(c) a magnet that generates a magnetic field and magnetic field gradient in said channel to alter passage of said one or more first analytes.

95. The system of claim 94, wherein said one or more second analytes comprise enucleated red blood cells.

96. The system of claim 94, wherein said one or more first analytes comprise nucleated red blood cells.

97. The system of claim 94, wherein said one or more first analytes comprise fetal nucleated red blood cells.

98. The system of claim 94, wherein said one or more first analytes comprise fetal hemoglobin, adult hemoglobin, methemoglobin, myoglobin, or a cytochrome.

99. The system of claim 94, further comprising a reservoir containing a deoxygenating agent coupled to said array of obstacles or said channel.

100. The system of claim 94, further comprising a reservoir containing a probe for specifically binding said one or more first analytes or components thereof.

101. The system of claim 100, wherein said probe is a nucleic acid probe or an antibody probe.

102. The system of claim 94, wherein said magnetic field is between 0.5 and 5.0 Tesla.

103. The system of claim 94, wherein said magnetic field gradient is between 100 Tesla/m and 1,000,000 Tesla/m.

104. The system of claim 94, wherein the passage of said one or more first analytes is altered based on an intrinsic magnetic property.

105. A system comprising (a) a flow-through channel comprising a two dimensional array of obstacles that selectively directs one or more first analytes having a hydrodynamic size greater than a critical size in a first direction towards a first outlet and one or more second analytes having a hydrodynamic size less than a critical size in a second direction towards a second outlet;

and (b) a magnet that generates a magnetic field and magnetic field gradient to alter the passage of said one or more first analytes.

106. The system of claim 105, wherein said one or more first analytes comprise fetal nucleated red blood cells.

107. The system of claim 105, wherein the passage of said one or more first analytes is altered based on the presence of hemoglobin.

108. The system of claim 105, wherein said magnetic field is between 0.5 and 5.0 Tesla.

109. The system of claim 105, wherein said magnetic field gradient is between 100 Tesla/m and 1,000,000 Tesla/m.