Droplet Assembly by 3D Printing

• US 20160136888A1
• Filed: 12/06/2013
• Published: 05/19/2016
• Est. Priority Date: 12/07/2012
• Status: Active Grant

• Current Assignee: Oxford University Innovation Limited (University of Oxford)
• Original Assignee: Isis Innovation Limited
• Inventors: Bayley, John Hagan Pryce, Villar, Gabriel, Graham, Alexander D.
• Granted Patent: US 11,213,797 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 9/127 Liposomes
• CPC Class Codes: A61K 9/1277 Processes for preparing; Proliposomes
• CPC Class Codes: B01J 13/04 by physical processes, e.g. drying, spraying
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2400/0433 vibrational forces
• CPC Class Codes: B01L 3/0268 using pulse dispensing or spraying, eg. inkjet type, piezo actuated ejection of droplets from capillaries
• CPC Class Codes: B29C 64/112 using individual droplets, e.g. from jetting heads
• CPC Class Codes: B29C 64/393 for controlling or regulating additive manufacturing processes
• CPC Class Codes: B29C 69/00 Combinations of shaping techniques not provided for in a single one of main groups B29C39/00 - B29C67/00, e.g. associations of moulding and joining techniques; Apparatus therefore B29C48/001 takes precedence
• CPC Class Codes: B29K 2105/0058 Liquid or visquous
• CPC Class Codes: B29L 2031/753 Medical equipment; Accessories therefor bloodbags, medical bags B29L2031/7148; artificial eyes B29L2011/0008
• CPC Class Codes: B33Y 10/00 Processes of additive manufacturing
• CPC Class Codes: B33Y 30/00 Apparatus for additive manufacturing; Details thereof or accessories therefor
• CPC Class Codes: B33Y 50/02 for controlling or regulating additive manufacturing processes
• CPC Class Codes: B33Y 80/00 Products made by additive manufacturing
• CPC Class Codes: C09B 57/001 Pyrene dyes
• CPC Class Codes: C09B 67/0097 Dye preparations of special physical nature; Tablets, films, extrusion, microcapsules, sheets, pads, bags with dyes

Abstract

The invention relates to an apparatus for producing a droplet assembly, which apparatus comprises a droplet generator. A process for producing a droplet assembly, using an apparatus comprising a droplet generator is also described. The invention also relates to droplet assemblies comprising a plurality of droplets. Various uses of the droplet assemblies are also described.

Claims

1. An apparatus for producing a droplet assembly, which apparatus comprises:

at least one droplet generator;

a container which is moveable relative to the at least one droplet generator; and

a control unit, which control unit is adapted to control the dispensing of droplets from the at least one droplet generator and the movement of the container relative to the at least one droplet generator,

wherein the apparatus is adapted to produce a droplet assembly which comprises a plurality of droplets, wherein each of said droplets comprises (i) a droplet medium, and (ii) an outer layer of amphipathic molecules around the surface of the droplet medium, wherein the droplet medium is an aqueous medium or a hydrophobic medium, and wherein at least one of said droplets contacts another of said droplets to form a layer of said amphipathic molecules as an interface between the contacting droplets.

2. An apparatus according to claim 1 which is adapted to produce a said droplet assembly wherein each of said droplets contacts another of said droplets to form a layer of said amphipathic molecules as an interface between the contacting droplets.

3. An apparatus according to claim 1 wherein the or each layer of said amphipathic molecules which is an interface between contacting droplets is a bilayer of said amphipathic molecules.

11. An apparatus according to claim 1 wherein the container contains a bulk medium, wherein:

when the droplet medium is an aqueous medium, the bulk medium is a hydrophobic medium, and

when the droplet medium is a hydrophobic medium, the bulk medium is an aqueous medium.

12. An apparatus according to claim 11 wherein the droplet generator contains said droplet medium.

13. An apparatus according to claim 12 wherein the droplet medium is an aqueous medium.

15. An apparatus according to claim 13 wherein the container contains a hydrophobic medium and amphipathic molecules.

18. An apparatus according to claim 1 wherein the or each droplet generator is adapted to dispense droplets having a diameter of equal to or less than 1 mm or having a volume of equal to or less than 2 μ

L.

23. An apparatus according to claim 1 wherein the apparatus comprises a plurality of said droplet generators.

24. An apparatus according to claim 23 wherein the droplet medium is an aqueous medium and wherein a first droplet generator comprises a first aqueous medium and wherein a second droplet generator comprises a second aqueous medium, wherein the first and second aqueous media are different, or wherein a first droplet generator is adapted to dispense droplets having a first diameter, and a second droplet generator is adapted to dispense droplets having a second diameter, wherein the first and second diameters are different.

29. An apparatus according to claim 1 wherein the control unit is adapted to coordinate (a) the movement of the container relative to the or each droplet generator and (b) the dispensing of the droplets, to create a said droplet assembly which comprises at least one layer of droplets, wherein each of said droplets comprises (i) a droplet medium and (ii) an outer layer of amphipathic molecules around the surface of the droplet medium, wherein the droplet medium is an aqueous medium or a hydrophobic medium and wherein each droplet in the layer contacts at least one other droplet in the layer to form a layer of said amphipathic molecules as an interface between contacting droplets.

32. An apparatus according to claim 1 wherein the control unit is adapted to coordinate (a) the movement of the container relative to the or each droplet generator and (b) the dispensing of the droplets, to create a said droplet assembly which comprises a plurality of droplets, wherein each of said droplets comprises (i) an aqueous medium, and (ii) an outer layer of amphipathic molecules around the surface of the aqueous medium, and wherein each of said droplets contacts another of said droplets to form a bilayer of said amphipathic molecules as an interface between the contacting droplets,

wherein the plurality of droplets comprises a first region of said droplets and a second region of said droplets, wherein each droplet in the first region contacts at least one other droplet in the first region to form a bilayer of said amphipathic molecules as an interface between the contacting droplets, and each droplet in the second region contacts at least one other droplet in the second region to form a bilayer of said amphipathic molecules as an interface between the contacting droplets,

wherein the aqueous medium of the droplets in the first region is different from the aqueous medium of the droplets in the second region, and/or wherein the composition of the bilayers between droplets in the first region is different from the composition of the bilayers between droplets in the second region.

33. An apparatus according to claim 32 wherein:

the aqueous medium of the droplets in the first region comprises a membrane protein and the aqueous medium of the droplets in the second region does not comprise said membrane protein;

the aqueous media of the droplets in the first and second regions have different concentrations of a membrane protein;

the bilayers between droplets in the first region further comprise a membrane protein, and the bilayers between droplets in the second region either do not comprise said membrane protein or have a lower concentration of said membrane protein than the bilayers between droplets in the first region;

the aqueous media of the droplets in the first and second regions have different osmolarities;

or

the aqueous media of the droplets in the first and second regions have different concentrations of a salt or buffer.

4-10. -10. (canceled)

14. (canceled)

16-17. -17. (canceled)

19-22. -22. (canceled)

25-28. -28. (canceled)

30-31. -31. (canceled)

34-37. -37. (canceled)

38. A process for producing a droplet assembly using an apparatus for producing the droplet assembly,

which droplet assembly comprises:

a plurality of droplets, wherein each of said droplets comprises;

(i) a droplet medium, and (ii) an outer layer of amphipathic molecules around the surface of the droplet medium, wherein the droplet medium is an aqueous medium or a hydrophobic medium, and wherein at least one of said droplets contacts another of said droplets to form a layer of said amphipathic molecules as an interface between the contacting droplets;

which apparatus comprises;

at least one droplet generator;

a container which is moveable relative to the at least one droplet generator; and

a control unit, which control unit is adapted to control the dispensing of droplets from the at least one droplet generator and the movement of the container relative to the at least one droplet generator;

wherein said container of the apparatus contains a bulk medium, wherein;

when the droplet medium is an aqueous medium the bulk medium is a hydrophobic medium, and when the droplet medium is a hydrophobic medium the bulk medium is an aqueous medium;

which process comprises;

(a) a plurality of dispensing steps, wherein each dispensing step comprises dispensing a droplet of the droplet medium from a said droplet generator into the bulk medium, in the presence of amphipathic molecules, and thereby forming in the bulk medium a droplet which comprises (i) said droplet medium and (ii) an outer layer of amphipathic molecules around the surface of the droplet medium; and

(b) moving the container relative to the at least one droplet generator, to control the relative positioning of the droplets in the bulk medium.

39. A process according to claim 38 wherein the or each layer of said amphipathic molecules which is an interface between contacting droplets is a bilayer of said amphipathic molecules.

41. A process according to claim 38 wherein the control unit coordinates the dispensing steps, and the movement of the container relative to the at least one droplet generator, to produce the droplet assembly.

42. A process according to claim 38 wherein moving the container relative to the at least one droplet generator comprises:

moving the container relative to the at least one droplet generator to position at least one of said droplets adjacent to another of said droplets so that at least one of said droplets contacts another of said droplets to form a layer of said amphipathic molecules as an interface between the contacting droplets.

44. A process according to claim 38, wherein moving the container relative to the at least one droplet generator comprises:

moving the container relative to the at least one droplet generator to position each droplet adjacent to at least one other droplet, so that each of said droplets contacts another of said droplets to form a layer of said amphipathic molecules as an interface between contacting droplets.

47. A process according to claim 38 wherein the droplet medium is an aqueous medium and the bulk medium is a hydrophobic medium.

48. A process according to claim 38 for producing a droplet assembly using an apparatus for producing the droplet assembly,

which droplet assembly comprises:

a plurality of droplets, wherein each of said droplets comprises;

(i) an aqueous medium, and (ii) an outer layer of amphipathic molecules around the surface of the aqueous medium, and wherein at least one of said droplets contacts another of said droplets to form a bilayer of said amphipathic molecules as an interface between the contacting droplets;

which apparatus comprises;

at least one droplet generator;

a container which is moveable relative to the at least one droplet generator; and

a control unit, which control unit is adapted to control the dispensing of droplets from the at least one droplet generator and the movement of the container relative to the at least one droplet generator;

wherein said container of the apparatus contains a hydrophobic medium;

which process comprises;

(a) a plurality of dispensing steps, wherein each dispensing step comprises dispensing a droplet of an aqueous medium from a said droplet generator into the hydrophobic medium, in the presence of amphipathic molecules, and thereby forming in the hydrophobic medium a droplet which comprises (i) said aqueous medium and (ii) an outer layer of amphipathic molecules around the surface of the aqueous medium; and

(b) moving the container relative to the at least one droplet generator, to control the relative positioning of the droplets in the hydrophobic medium.

49. A process according to claim 47 wherein the process comprises:

(a) a first dispensing step, comprising dispensing a droplet of an aqueous medium from a said droplet generator into the hydrophobic medium, in the presence of amphipathic molecules, and thereby forming in the hydrophobic medium a first droplet, which first droplet comprises (i) said aqueous medium and (ii) an outer layer of amphipathic molecules around the surface of the aqueous medium;

(b) moving the container relative to the at least one droplet generator, to control the positioning of a second droplet in the hydrophobic medium relative to the first droplet in the hydrophobic medium; and

(c) a second dispensing step, comprising dispensing a droplet of an aqueous medium from a said droplet generator into the hydrophobic medium, in the presence of amphipathic molecules, and thereby forming in the hydrophobic medium a second droplet, which second droplet comprises (i) said aqueous medium and (ii) an outer layer of amphipathic molecules around the surface of the aqueous medium.

50. A process according to claim 49 wherein said (b) moving the container relative to the at least one droplet generator comprises:

moving the container relative to the at least one droplet generator so that the second droplet is positioned adjacent to the first droplet, so that the first and second droplets contact one another to form a bilayer of said amphipathic molecules as an interface between the contacting droplets.

51. A process according to claim 50 wherein the process further comprises:

(d) moving the container relative to the at least one droplet generator, to control the positioning of a further droplet in the hydrophobic medium relative to the other droplets in the hydrophobic medium; and

(e) a further dispensing step, comprising dispensing a droplet of an aqueous medium from a said droplet generator into the hydrophobic medium, in the presence of amphipathic molecules, and thereby forming in the hydrophobic medium a further droplet, which further droplet comprises (i) said aqueous medium and (ii) an outer layer of amphipathic molecules around the surface of the aqueous medium.

52. A process according to claim 51 wherein said (d) moving the container relative to the at least one droplet generator, comprises:

moving the container relative to the at least one droplet generator so that the further droplet is positioned adjacent to at least one other droplet in the hydrophobic medium, so that the further droplet contacts at least one other droplet in the hydrophobic medium to form a bilayer of said amphipathic molecules as an interface between the contacting droplets.

53. A process according to claim 51 which comprises at least 500 of said further dispensing steps (e).

57. A process according to claim 47 wherein the plurality of dispensing steps comprises dispensing steps which together produce a row or layer of said droplets in the hydrophobic medium, wherein each droplet in the row or layer contacts another droplet in the row or layer to form a bilayer of said amphipathic molecules as an interface between contacting droplets.

76. A process according to claim 38 wherein the droplet medium is an aqueous medium and the bulk medium is a hydrophobic medium and wherein the plurality of dispensing steps comprises:

a first set of dispensing steps, which together produce a first region of said droplets in the hydrophobic medium, wherein each droplet in the first region contacts at least one other droplet in the first region to form a bilayer of said amphipathic molecules as an interface between contacting droplets; and

a second set of dispensing steps, which together produce a second region of said droplets in the hydrophobic medium, wherein each droplet in the second region contacts at least one other droplet in the second region to form a bilayer of said amphipathic molecules as an interface between contacting droplets.

82. A process according to claim 76 wherein the aqueous medium of the droplets dispensed into the hydrophobic medium in said first set of dispensing steps further comprises a membrane protein, and wherein the aqueous medium of the droplets dispensed into the hydrophobic medium in said second set of dispensing steps does not comprise said membrane protein, or wherein the aqueous medium of the droplets dispensed into the hydrophobic medium in said first set of dispensing steps comprises a higher concentration of a membrane protein than the aqueous medium of the droplets dispensed into the hydrophobic medium in said second set of dispensing steps.

86. A process according to claim 82 wherein the bilayers of amphipathic molecules formed between the contacting droplets in the first region comprise said membrane protein, and the bilayers of amphipathic molecules formed between the contacting droplets in the second region do not comprise said membrane protein or comprise a lower concentration of said membrane protein than the bilayers of amphipathic molecules formed between the contacting droplets in the first region.

88. A process according to claim 82 wherein the membrane protein is a pump, channel or a pore, a receptor protein, a transporter protein, or a protein which effects cell recognition or a cell-to-cell interaction.

90. A process according to claim 76 wherein:

the aqueous medium of the droplets dispensed into the hydrophobic medium in said first set of dispensing steps has a first osmolarity, and the aqueous medium of the droplets dispensed into the hydrophobic medium in said second set of dispensing steps has a second osmolarity, wherein the first osmolarity is greater than the second osmolarity; and

droplets in the second region are disposed adjacent to droplets in the first region, so that droplets in the first region contact droplets in the second region to form bilayers of said amphipathic molecules as interfaces between the contacting droplets.

95. A process according to claim 90 which further comprises allowing water to transfer from the second region to the first region, to cause deformation of the droplet assembly.

96. A process according to claim 95 wherein the transfer causes the first and second regions to curve or fold.

99. A process for producing a droplet assembly according to claim 38 wherein the droplet medium is an aqueous medium and the bulk medium is a hydrophobic medium and wherein the apparatus comprises a first droplet generator and a second droplet generator, and the plurality of dispensing steps comprises:

at least one dispensing step which comprises dispensing a droplet of a first aqueous medium from the first droplet generator into the hydrophobic medium, in the presence of amphipathic molecules, and thereby forming in the hydrophobic medium a said droplet which comprises (i) said first aqueous medium and (ii) an outer layer of amphipathic molecules around the surface of the aqueous medium; and

at least one dispensing step which comprises dispensing a droplet of a second aqueous medium from the second droplet generator into the hydrophobic medium, in the presence of amphipathic molecules, and thereby forming in the hydrophobic medium a said droplet which comprises (i) said second aqueous medium and (ii) an outer layer of amphipathic molecules around the surface of the aqueous medium,

wherein the first aqueous medium and the second aqueous medium are the same or different.

102. A process according to claim 99 wherein the first and second aqueous media have different osmolarities, or the first and second aqueous media comprise different concentrations of a salt or buffer, the first aqueous medium comprises a membrane protein and the second aqueous medium does not comprise said membrane protein, or the droplet or droplets of the first aqueous medium, which are dispensed by the first droplet generator, have a different size from the droplet or droplets of the second aqueous medium, which are dispensed by the second droplet generator.

105. A process for producing a droplet assembly according to claim 38 wherein the droplet medium is an aqueous medium and the bulk medium is a hydrophobic medium, and wherein said bulk medium which is a hydrophobic medium is a drop of a hydrophobic medium.

106. A process for producing a droplet assembly according to claim 105 wherein the drop of the hydrophobic medium is within a second bulk medium which is an aqueous medium, wherein the container of the apparatus contains said second bulk medium which is an aqueous medium and the drop of hydrophobic medium, and the drop of the hydrophobic medium further comprises a peripheral layer of amphipathic molecules around the surface of the drop, as an interface between the drop and the second bulk medium.

112. A droplet assembly which is obtainable by a process as defined in claim 38.

127. A method of using a droplet assembly, the method comprising using a droplet assembly as defined in claim 112 in synthetic biology, as a drug-delivery vehicle, in tissue engineering, or in material science and engineering.

40. (canceled)

43. (canceled)

45-46. -46. (canceled)

54-56. -56. (canceled)

58-75. -75. (canceled)

77-81. -81. (canceled)

83-85. -85. (canceled)

87. (canceled)

89. (canceled)

91-94. -94. (canceled)

97-98. -98. (canceled)

100-101. -101. (canceled)

103-104. -104. (canceled)

107-111. -111. (canceled)

113. A droplet assembly which comprises a plurality of droplets, wherein each of said droplets comprises (i) an aqueous medium, and (ii) an outer layer of amphipathic molecules around the surface of the aqueous medium, and wherein each of said droplets contacts another of said droplets to form a bilayer of said amphipathic molecules as an interface between the contacting droplets,

wherein the plurality of droplets comprises a first region of said droplets and a second region of said droplets, wherein each droplet in the first region contacts at least one other droplet in the first region to form a bilayer of said amphipathic molecules as an interface between the contacting droplets, and each droplet in the second region contacts at least one other droplet in the second region to form a bilayer of said amphipathic molecules as an interface between the contacting droplets,

wherein the aqueous medium of the droplets in the first region has a first osmolarity and the aqueous medium of the droplets in the second region has a second osmolarity, wherein the first osmolarity is different from the second osmolarity.

114. A droplet assembly according to claim 113 wherein the ratio of the first osmolarity to the second osmolarity is from 2:

1 to 50;

1.

115. A droplet assembly according to claim 113 wherein the aqueous medium of the droplets in the first region is an aqueous solution of a salt, which salt has a first concentration in the aqueous solution, and the aqueous medium of the droplets in the second region is an aqueous solution of the same salt, which salt has a second concentration in the aqueous solution, wherein the first concentration is different from the second concentration.

116. A droplet assembly according to claim 115, wherein the ratio of the first concentration to the second concentration is from 2:

1 to 50;

1.

126. A droplet assembly according to claim 113 wherein the number of said droplets in the plurality of droplets is at least 10,000.

117-120. -120. (canceled)

121. A droplet assembly which comprises a plurality of droplets, wherein each of said droplets comprises (i) an aqueous medium, and (ii) an outer layer of amphipathic molecules around the surface of the aqueous medium, and wherein each of said droplets contacts another of said droplets to form a bilayer of said amphipathic molecules as an interface between the contacting droplets, wherein the plurality of droplets defines a shell around a volume within the droplet assembly that does not comprise said droplets.

124. A droplet assembly according to claim 121 wherein the shell defined by said droplets is a curved structure, substantially cylindrical, substantially ring-shaped, substantially spherical, or substantially hemispherical, and encloses said volume within the droplet assembly.

122-123. -123. (canceled)

125. (canceled)

128-130. -130. (canceled)