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1. (WO2015175344) SYSTÈME ET APPAREIL PERMETTANT D'ISOLER OU D'ENRICHIR DES AGENTS PAR FLOTTATION
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CLAIMS

What is claimed is:

1. An apparatus for isolation or enrichment of biological agents from a sample comprising:

a sample container comprising:

(a) an inner volume to hold a sample and a plurality of microbubbles;

(b) a tapered end and an open end;

(c) a plunger inserted into said open end; and

(d) a tip of said tapered end forming an acute angle relative to the horizontal axis of said sample container,

wherein said plurality of microbubbles float one or more biological agents of said sample and wherein said plunger moves towards said tip to isolate or enrich said one or more biological agents from said sample.

2. The apparatus of claim 1, wherein said acute angle is between about 5 degrees and about 40 degrees.

3. The apparatus of claim 1, wherein said acute angle is between about 5 degrees and about 10 degrees.

4. The apparatus of claim 1, wherein said acute angle is about 35 degrees.

5. The apparatus of claim 1, further comprising a container spout directionally orienting one or more droplets that form from said tip comprising said one or more biological agents.

6. The apparatus of claim 5, further comprising a collection container to collect the one or more droplets.

7. The apparatus of claim 6, wherein said collection container is adjacent to the outer surface of the sample container.

8. The apparatus of claim 7, wherein said collection container is mounted onto the outer surface of the sample container close to the bottom end of the container spout.

9. An apparatus for isolation or enrichment of biological agents from a sample comprising:

a sample container comprising:

(a) an inner volume to hold a sample and a plurality of microbubbles;

(b) a tapered end and an open end;

(c) a plunger inserted into said open end; and

(d) a tip of said tapered end perpendicular relative to the horizontal axis of said sample container,

wherein said plurality of microbubbles float one or more biological agents of said sample and wherein said plunger moves towards said tip to isolate or enrich said one or more biological agents from said sample.

10. The apparatus of claims 1 or 9, wherein the plunger is operated manually by a user.

11. The apparatus of claims 1 or 9, further comprising a drive screw mechanically coupled to said plunger, wherein rotation of said drive screw is mechanically converted to linear translation of said plunger.

12. The apparatus of claim 11, further comprising a container mount to house the drive screw.

13. The apparatus of claim 12, wherein a bottom flange of said sample container is secured to said container mount by inserting said bottom flange into a locking flange slot on said container mount.

14. The apparatus of claim 11, wherein said drive screw is operated manually by a user.

15. The apparatus of claim 11, wherein said drive screw is operated automatically by a drive motor.

16. The apparatus of claim 11, wherein said drive screw is operated automatically by a drive motor remotely controlled by a microprocessor.

17. The apparatus of claim 15, further comprising an apparatus enclosure to house said drive motor.

18. The apparatus of claim 11, further comprising a plunger adapter joint and a plunger engagement adapter, wherein said plunger adapter joint is adjacent to said plunger and said plunger engagement adapter, and said plunger engagement adapter is adjacent to said plunger adapter joint and said drive screw.

19. The apparatus of claim 12, wherein said container mount further comprising a sensor depressor and sensor mount.

20. The apparatus of claim 12, wherein said container mount further comprises a wire port.

21. The apparatus of claim 12, wherein said container mount further comprises a screw port.

22. The apparatus of claims 1 or 9, wherein said sample container is formed of a material to provide said inner volume of said sample container hydrophilicity.

23. The apparatus of claims 1 or 9, wherein said sample container is formed of a material to provide said inner volume of said sample container hydrophobicity.

24. The apparatus of claim 22, wherein said material is glass.

25. The apparatus of claims 1 or 9, wherein a hydrophilic layer is adjacent thereto said inner volume of said sample container.

26. The apparatus of claims 1 or 9, wherein a hydrophobic layer is adjacent thereto said tapered end of said inner volume of said sample container.

27. The apparatus of claims 1 or 9, wherein a hydrophilic layer is adjacent thereto said inner volume of said sample container and a hydrophobic layer is adjacent thereto said tapered end of said inner volume.

28. The apparatus of claims 1 or 9, wherein said sample container is formed of a material to provide hydrophilicity to said inner volume and a hydrophobic layer is adjacent thereto said tapered end of said inner volume.

29. The apparatus of claims 25 or 27, wherein said hydrophilic layer is bovine serum albumin.

30. The apparatus of any one of claims 26, 27, or 28 wherein said hydrophobic layer is SURFASIL™.

31. The apparatus of any one of claims 26, 27, or 28 wherein said hydrophobic layer is paraffin, polytetrafluoroethylene, poloaxmer, or combinations thereof.

32. The apparatus of claims 1 or 9, wherein said sample container holds a sample volume of between about lmL and about lOmL.

33. The apparatus of claims 1 or 9, wherein said sample container holds a sample volume of between about lOmL and about 50mL.

34. The apparatus of claims 1 or 9, wherein said sample container holds a sample volume of between about 50mL and about 200mL.

35. The apparatus of claims 1 or 9, wherein said sample is diluted.

36. The apparatus of claims 1 or 9, wherein said sample is tissue, blood, bone marrow, urine, saliva, cerebrospinal fluid, seminal fluid, sputum, stool, joint fluid, lymph, amniotic fluid, bile, ascites, or pleural effusion, or combinations thereof.

37. The apparatus of claims 1 or 9, wherein said sample is umbilical cord blood.

38. The apparatus of claims 1 or 9, wherein said sample is circulating blood.

39. The apparatus of claims 1 or 9, wherein said sample is tumor tissue.

40. The apparatus of claims 1 or 9, wherein said sample is fluid within and surrounding a tumor tissue.

41. The apparatus of claims 1 or 9, wherein said biological agents are cells.

42. The apparatus of claim 41, wherein said cells are circulating tumor cells (CTCs).

43. The apparatus of claim 41, wherein said cells are bone marrow cells.

44. The apparatus of claim 41, wherein said cells are fetal cells from a mother's blood.

45. The apparatus of claim 41, wherein a ratio of said cells to said plurality of microbubbles is about 1 : 100.

46. The apparatus of claim 41, wherein a ratio of said cells to said plurality of

microbubbles is about 1 :500.

47. The apparatus of claim 41, wherein a ratio of said cells to said plurality of

microbubbles is about 1 : 1000.

48. The apparatus of claims 1 or 9, wherein said biological agents isolated or enriched are less than 0.5% of the total sample.

49. The apparatus of claim 36, wherein said biological agents isolated or enriched are less than 0.1% of the total sample.

50. The apparatus of claim 49, wherein said biological agents isolated or enriched are less than 0.01% of the total sample.

51. The apparatus of claim 50, wherein said biological agents isolated or enriched are less than 0.001% of the total sample.

52. The apparatus of claims 1 or 9, wherein said microbubbles are lipid shell microbubbles, albumin shell microbubbles, polymer shell microbubbles, lipopolymer shell microbubbles or glass microbubbles, or combinations thereof.

53. The apparatus of claims 52, wherein the shell of each microbubble comprises lipid, protein, polymer, lipopolymer, or glass or combinations thereof.

54. The apparatus of claims 1 or 9, wherein said plurality of microbubbles have an outer diameter of between about 1 micron and about 10 microns.

55. The apparatus of claims 1 or 9, wherein said plurality of microbubbles have an outer diameter of between about 2 microns and about 8 microns.

56. The apparatus of claims 1 or 9, wherein said plurality of microbubbles have an outer diameter of between about 2 microns and about 20 microns.

57. The apparatus of claims 1 or 9, wherein a plurality of selection markers are adjacent to the outer surface of the plurality of microbubbles.

58. The apparatus of claim 57, wherein said plurality of selection markers bind said outer surface by nucleophilic conjugate addition.

59. The apparatus of claim 58, wherein said plurality of selection markers bind said outer surface by Michael addition.

60. The apparatus of claim 57, wherein said plurality of selection markers per microbubble is between about 200,000 and about 500,000.

61. The apparatus of claim 57, wherein a surface density of the plurality of selection markers adjacent to the outer surface of a single microbubble of the plurality of microbubbles is between about 3,000 and about 6,000 selection markers per square micron.

62. The apparatus of claim 57, wherein a surface density of the plurality of selection markers adjacent to the outer surface of a single microbubble of the plurality of microbubbles is between about 1,000 and about 3,000 selection markers per square micron.

63. The apparatus of claim 57, wherein a surface density of the plurality of selection markers adjacent to the outer surface of a single microbubble of the plurality of microbubbles is less than about 6,000 selection markers per square micron.

64. The apparatus of claim 57, wherein a surface density of the plurality of selection markers adjacent to the outer surface of a single microbubble of the plurality of microbubbles is less than about 3,000 selection markers per square micron.

65. The apparatus of claim 60, wherein said plurality of selection markers per microbubble is between about 300,000 and about 400,000.

66. The apparatus of claim 57, wherein said plurality of selection markers comprise a first set and a second set.

67. The apparatus of claim 66, wherein each of said plurality of microbubbles comprise said first set and said second set of selection markers.

68. The apparatus of claim 66, wherein a first set of said plurality of microbubbles comprise said first set of selection markers and a second set of said plurality of microbubbles comprise said second set of selection markers.

69. The apparatus of claim 41, wherein said cells attach said plurality of microbubbles by receptor ligand interactions.

70. The apparatus of claim 57, wherein said one or more selection markers bind cell surface markers.

71. The apparatus of claim 57, wherein said one or more selection markers are different.

72. The apparatus of claim 57, wherein said one or more selection markers are antibodies.

73. The apparatus of claim 57, wherein said one or more selection markers are molecules that bind tumor markers.

74. The apparatus of claim 57, wherein said one or more selection markers are molecules that bind stem cell markers.

75. The apparatus of claim 57, wherein said one or more selection markers are molecules that bind cell surface marker, EpCAM.

76. The apparatus of claim 57, wherein said one or more selection markers are molecules that bind cell surface marker, CD 133.

77. The apparatus of claim 57, wherein said one or more selection markers are molecules that bind cell surface marker, CD34.

78. The apparatus of claim 70, wherein at least about 80% of said cells bind at least one of said plurality of microbubbles.

79. The apparatus of claim 70, wherein at least about 85% of said cells bind at least one of said plurality of microbubbles.

80. The apparatus of claim 70, wherein at least about 90% of said cells bind at least one of said plurality of microbubbles.

81. The apparatus of claim 70, wherein at least about 95% of said cells bind at least one of said plurality of microbubbles.

82. The apparatus of claims 1 or 9, wherein a biological agent of said sample floats when at least about 2 microbubbles are adjacent thereto.

83. The apparatus of claims 1 or 9, wherein a biological agent of said sample floats when at least about 3 microbubbles are adjacent thereto.

84. The apparatus of claims 1 or 9, wherein a biological agent of said sample floats when between about 2 to about 20 microbubbles are adjacent thereto.

85. The apparatus of claims 1 or 9, wherein said apparatus is compatible with downstream assays.

86. The apparatus of claim 85, wherein said downstream assays comprise cell staining, molecular analysis, or cell culture, or combinations thereof.

87. The apparatus of claims 1 or 9, wherein said apparatus is sterile.

88. The apparatus of claims 1 or 9, further comprising a sterile liquid in said inner volume to enhance viability of said sample.

89. The apparatus of claims 1 or 9, further comprising a sterile liquid in said inner volume to cause said plurality of microbubbles to float.

90. The apparatus of claim 89, wherein the density of said liquid is between about 0.9 and about 1.1 grams per cubic centimeter.

91. The apparatus of claims 1 or 9, wherein said apparatus is used in a clinical or laboratory setting.

92. The apparatus of claims 1 or 9, wherein said apparatus is used in a residential setting.

93. The apparatus of claims 1 or 9, wherein said apparatus is utilized for personal use.

94. The apparatus of claim 15, further comprising a bottom stop sensor, a stepper motor, a top drip detector, a power supply, a microcontroller board, or combinations thereof.

95. A method for isolation or enrichment of biological agents from a sample comprising:

(a) obtaining a sample and a plurality of microbubbles;

(b) adding said sample and said plurality of microbubbles to an inner volume of a

sample container;

(c) orienting said tapered end of said sample container away from ground wherein said plurality of microbubbles float one or more biological agents towards a tip of said tapered end; and

(d) moving a plunger from an open end of said sample container towards said tapered end to isolate or enrich one or more biological agents from said sample.

96. The method of claim 95, further comprising disrupting said plurality of microbubbles after (d).

97. The method of claim 96, wherein ambient pressure is increased to disrupt said plurality of microbubbles.

98. The method of claim 97, wherein ambient pressure is increased to between about 2 atm and about 4 atm for between about 2 minutes and about 10 minutes to disrupt said plurality of microbubbles.

99. The method of claim 98, wherein ambient pressure is increased to about 3 atm for about 5 minutes to disrupt said plurality of microbubbles.

100. The method of claim 95, further comprising incubating said plurality of microbubbles and said sample in said inner volume for at least about 1 minute before (c) or (d).

101. The method of claim 100, further comprising incubating said plurality of microbubbles and said sample in said inner volume for at least about 5 minutes before (c) or (d).

102. The method of claim 101, further comprising incubating said plurality of microbubbles and said sample in said inner volume for at least about 10 minutes before (c) or (d).

103. The method of claim 95, further comprising sterilizing said inner volume of said sample container prior to adding of (b).

104. A kit for isolation or enrichment of biological agents from a sample comprising:

(a) one or more sample containers;

(b) an apparatus enclosure;

(c) a container mounts;

(d) one or more selection markers;

(e) components to form microbubbles;

(f) instructions for forming microbubbles; and

(g) instructions for using said sample container.

105. A kit for isolation or enrichment of biological agents from a sample comprising:

(a) one or more sample containers;

(b) an apparatus enclosure;

(c) a container mount;

(d) microbubbles with one or more selection markers adjacent thereto; and

(e) instructions for using said sample container.

106. A kit for isolation or enrichment of biological agents from a sample comprising:

(a) one or more sample containers;

(b) one or more selection markers;

(c) components to form microbubbles;

(d) instructions for forming microbubbles; and

(e) instructions for using said sample container.

107. The apparatus of claims 1 or 9, wherein said apparatus is used in a remote geographical area.

108. The apparatus of claims 1 or 9, wherein said apparatus is used in an impoverished or low-income area.

109. The apparatus of claims 1 or 9, wherein said apparatus is used in the absence of sterilization equipment.

110. The apparatus of claims 1 or 9, wherein said apparatus is used in the absence of centrifugation equipment.

111. The apparatus of claims 1 or 9, wherein said apparatus is used by a clinician.

112. The apparatus of claims 1 or 9, wherein said apparatus is used by a researcher.

113. The apparatus of claims 1 or 9, wherein said apparatus is used by a lay person.

114. The apparatus of claims 1 or 9, wherein said apparatus is used by a consumer.

115. The apparatus of claims 1 or 9, wherein said apparatus is used for cord blood banking,

116. The apparatus of claims 1 or 9, wherein said apparatus is used for isolation of immune cells for immunotherapy in cancer treatment.

117. The apparatus of claims 1 or 9, wherein said apparatus is used for isolation of hematopoietic stem cells from umbilical cord blood following a live birth.

118. The apparatus of claims 1 or 9, wherein said apparatus is used for isolation of one or more narcotics from a blood sample or urine sample.

119. The apparatus of claims 1 or 9, wherein said apparatus is used for depletion of one or more biological agents from a sample.

120. The apparatus of claims 1 or 9, wherein said apparatus is used for depletion of one or more non-biological agents from a sample.

121. An apparatus for isolation or enrichment of agents from a sample comprising:

a sample container comprising:

(e) an inner volume to hold a sample and a liquid;

(f) a tapered end and an open end;

(g) a plunger inserted into said open end; and

(h) a tip of said tapered end forming an acute angle relative to the horizontal axis of said sample container,

wherein said liquid floats one or more agents of said sample and wherein said plunger moves towards said tip to isolate or enrich said one or more agents from said sample.